RU2851685C2 - Compositions of antibodies for destruction of biofilms - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: methods and compositions for destroying biofilms in vitro and in vivo are proposed.

EFFECT: new antibodies capable of specifically binding to the head region of the DNABII peptide.

23 cl, 15 dwg, 9 tbl, 11 ex

Description

Cross-reference to related applications

This application claims priority under 35 U.S.C. § 119(e) to prior U.S. application No. 63/033,109, filed June 1, 2020, and U.S. application No. 62/871,457, filed July 8, 2019, the contents of each of which are incorporated herein by reference in their entireties.

Statement of state support

This invention was made with federal support under Grant No. R01 DC011818 from the National Institute on Deafness and Other Communication Disorders (NIDCD) of the National Institutes of Health (NIH). The government retains certain rights in the invention.

Field of technology to which the invention relates

The present invention generally relates to methods and compositions for reducing and/or eliminating bacterial biofilms.

Prior art

The DNABII protein family is naturally present outside bacterial cells and promotes biofilm formation. At least one DNABII protein is found in all known eubacteria. While these proteins elicit a strong innate and adaptive immune response, hosts fail to naturally produce protective antibodies against members of this family following infection. The primary problem with bacterial biofilms is the inability of the host immune system and/or antibiotics and other antimicrobial agents to access bacteria protected by the biofilm.

Biofilms are also found in industrial settings. For example, biofilms are associated with a wide range of oil process problems, from production fields to fuel tanks at gas stations. In the field, sulfate-reducing biofilm bacteria produce hydrogen sulfide (hydrogen sulfide-containing crude). In process pipelines, biofilm activity leads to the formation of slime, which obstructs filters and orifices. Biofilm and biofilm organisms also cause corrosion of pipelines and refining equipment. These problems can manifest themselves at all oil and gas production facilities, to the point that fouling and corrosive biofilm organisms are even found on the surfaces of final product storage tanks.

Biofilms are involved in a wide range of water processes, both domestic and industrial. They can grow on the surface of process equipment and interfere with its operation, for example, by impairing heat transfer or clogging filters and membranes. Biofilms growing on cooling tower packing can add enough mass to cause packing failure. Biofilms cause corrosion even in highly specialized stainless steels. Biofilms in water processes can reduce the value of the final product, for example, biofilm contamination in the papermaking process or the attachment of even a single cell to a silicon chip. Biofilms growing in drinking water distribution systems can harbor potential pathogens, corrosive organisms, or bacteria that degrade the aesthetic quality of the water. In households, biofilms are found in or on any surface that supports microbial growth, such as drains, food preparation surfaces, toilets, and swimming pools and spas.

Thus, there is a need to break through the protective barrier of biofilms to treat or kill associated bacterial infections and remove them from surfaces and water systems.

The essence of the invention

In bacterial cells, DNABII proteins are DNA-binding proteins that necessarily bend DNA substrates upon binding. Similarly, DNA already in a bent conformation is an ideal substrate, since the energy required for bending is unnecessary.

The DNABII family is a member of a class of proteins called nucleoid-associated proteins (NAPs), bacterial proteins that partially form the intracellular bacterial nucleoid (Browning et al. (2010) Curr. Opin. Microbiol. 13:773–780). Furthermore, this family is ubiquitous and is expressed by virtually all eubacteria. All characterized members of the family to date function as either homodimers or heterodimers of subunits. The family is divided into two types: HU (histone-like protein) and IHF (integration host factor protein), with B. cenocepacia being able to express both proteins (strain J2315 genes: BCAL3530, hupA; BCAL1585, hupB; BCAL1487, ihfA and BCAL2949, ihfb), while many other bacteria also express both HU and IHF. The main difference between these family members is that HU binds DNA in a sequence-independent manner, while IHF binds a consensus sequence (WATCAANNNNTTR, where W is A or T and R is a purine, SEQ ID NO: 28) conserved across genera (Swinger et al. (2004) Curr. Opin. Struct. Biol. 14:28–35). All DNABII proteins bind to DNA and bend it significantly, i.e., E. coli IHF can bend DNA into a virtual U-turn (Rice et al. (1996) Cell 87: 1295–1306). In addition, all members of the family prefer pre-bent or curved DNA structures, i.e., Holliday junctions, a cruciform structure located at the center of DNA recombination. In fact, DNABII proteins function as accessory factors facilitating all functions of intracellular DNA, including gene expression, recombination, repair, and replication (Swinger et al. (2004) Curr. Opin. Struct. Biol. 14:28–35).

DNABII family proteins are found outside bacterial cells in biofilm formation. The researchers demonstrated that these proteins are actually associated with extracellular DNA at critical branched junctions.

Thus, the present application provides an antibody or antigen-binding fragment thereof, which comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising or essentially consisting of or additionally consisting of a sequence of amino acids (aa) 25-144 of SEQ ID NO: 13 or an equivalent thereof, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising or essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-132 of SEQ ID NO: 14 or an equivalent thereof. Furthermore, the present application provides an antibody or antigen-binding fragment thereof, which comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising or essentially consisting of a sequence selected from the group of aa 25-144 of SEQ ID NO: 13, 24 or 26, or the equivalent of each of them, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising or essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-132 of SEQ ID NO: 14 or 25, aa 21-126 of SEQ ID NO: 27, or the equivalent of each of them. In some embodiments, the antibody or fragment thereof binds to a DNABII peptide (such as the head region of the DNABII peptide, including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region and/or the IhfA5-mIhfB4NTHI chimeric head peptide and/or the tail region of the DNABII peptide, including, but not limited to, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI chimeric head peptide or the IhfA3-IhfB2NTHI chimeric tail peptide.

Also provided in the present application is an antibody or antigen-binding fragment thereof, which comprises or consists essentially of or additionally consists of: a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising or consisting essentially of or additionally consisting of a sequence selected from the group aa 25-144 of SEQ ID NO: 13 or 24, or an equivalent of each of them, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising or consisting essentially of or additionally consisting of a sequence selected from the group aa 21-132 of SEQ ID NO: 14 or 25, or an equivalent of each of them. In some embodiments, the antibody or fragment thereof binds to a DNABII peptide (such as the head region of the DNABII peptide, including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI chimeric head peptide.

Also additionally provided is an antibody or an antigen-binding fragment thereof, which comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising or essentially consisting of or additionally consisting of a sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 24 or 26, or an equivalent of each of them, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising or essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-132 of SEQ ID NO: 7-9 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them. In some embodiments, the antibody or fragment thereof binds to a DNABII peptide (such as the head region of the DNABII peptide, including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region and/or the IhfA5-mIhfB4NTHI chimeric head peptide and/or the tail region of the DNABII peptide, including, but not limited to, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI chimeric head peptide or the IhfA3-IhfB2NTHI chimeric tail peptide.

Also provided is an antibody or antigen-binding fragment thereof, which comprises or essentially consists of or additionally consists of a heavy chain (HC) comprising or essentially consists of or additionally consists of a sequence selected from the group of aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or the equivalent of each of them, and/or a light chain (LC) comprising or essentially consists of or additionally consists of a sequence selected from the group of aa 21-239 of SEQ ID NO: 7-9, 14 or 25, 21-233 of SEQ ID NO: 10-12 or 27, or the equivalent of each of them. In some embodiments, the antibody or fragment thereof binds to a DNABII peptide (such as the head region of the DNABII peptide, including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region and/or the IhfA5-mIhfB4NTHI chimeric head peptide and/or the tail region of the DNABII peptide, including, but not limited to, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI chimeric head peptide or the IhfA3-IhfB2NTHI chimeric tail peptide.

Additionally provided is an antibody or fragment thereof that comprises or essentially consists of or additionally consists of any one, any two, or all three CDR sequences selected from the group of SEQ ID NOs: 1-6, 13, 24, or 26, or an equivalent of each of them; and/or any one, or any two, or all three CDR sequences selected from the group of SEQ ID NOs: 7-12, 14, 25, or 27, or an equivalent of each of them. In some embodiments, the antibody or fragment thereof binds to a DNABII peptide (such as the head region of the DNABII peptide, including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region and/or the IhfA5-mIhfB4NTHI chimeric head peptide and/or the tail region of the DNABII peptide, including, but not limited to, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI chimeric head peptide or the IhfA3-IhfB2NTHI chimeric tail peptide.

Non-limiting examples of antigen-binding fragments are selected from Fab, F(ab’)2, Fab’, scFv or Fv.

In another aspect, the present application provides a Fab fragment of an antibody, wherein the antibody specifically binds to the head region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). Also provided is a fragment of the antibody described in the present application. In one embodiment, the fragment is an antigen-binding fragment selected from the group of Fab, F(ab')2, Fab', scFv, or Fv. In another embodiment, the fragment specifically binds to the head region of a DNABII peptide.

In addition, formulations containing the Fab fragment are offered.

In another aspect, the antibody or fragment thereof is modified. Non-limiting examples of modifications include PEGylation, PEG mimetic modification, polysialylation, HESylation, or glycosylation.

Antibodies and antigen-binding fragments may be labeled with a detectable label or contain a detectable label and/or a purification marker.

Also provided in the present application are polypeptides comprising or consisting essentially of or consisting of an antigen-binding complementarity determining region (CDR) belonging to an antibody or antigen-binding fragment or region, as described above. The CDR can be selected from the group CDR1, CDR2, or CDR3.

CDRs may be labeled with a detectable label or contain a detectable label and/or a purification marker.

Further provided are isolated polypeptides, wherein the polypeptides comprise, consist essentially of, or additionally consist of an amino acid sequence selected from the group of SEQ ID NOs: 1-14 or 24-27; amino acids (aa) 25-144 of SEQ ID NOs: 1-6, 13, 24, or 26; 21-132 aa of SEQ ID NOs: 7-9, 14, or 25; 21-126 aa of SEQ ID NOs: 10-12 or 27; or an equivalent of each thereof. Also provided is an isolated polypeptide comprising or alternatively consisting essentially of or additionally consisting of one or more amino acid sequences of antibodies or fragments thereof, as disclosed herein. The polypeptides may be labeled with a detectable label and/or contain a detectable label and/or a purification marker. In another embodiment, the polypeptides additionally contain a signal peptide.

Further provided are isolated polynucleotides encoding antibodies or antigen-binding fragments thereof, or CDRs of antigen-binding fragments or antibodies, as well as polypeptides of the present invention or equivalents of each, which are optionally operably linked to a promoter and/or enhancer element. The polynucleotides may be labeled with a detectable label and/or contain a detectable label and/or a purification marker. The polynucleotides may further comprise a signal peptide polynucleotide sequence located upstream of the immunoglobulin variable domain of the antibody. They may be contained within a vector and/or a host cell. Thus, further provided is a vector comprising or alternatively consisting essentially of one or more isolated polynucleotides as disclosed herein, or further consisting of one or more isolated polynucleotides. Such a vector may be a plasmid or viral vector, optionally selected from the group consisting of a retroviral vector, a lentiviral vector, an adenoviral vector, and an adeno-associated virus vector. Also provided is a host cell comprising one or more polynucleotides and/or vectors as disclosed herein.

Further provided are compositions comprising or alternatively consisting of or additionally consisting of a carrier and one or more compounds consisting of an antibody or fragment thereof, CDRs, polypeptides, isolated polynucleotides, vectors, or a host cell. In one embodiment, the carrier is a pharmaceutically acceptable carrier. Additionally or alternatively, the carrier is a solid-phase carrier or a solid-phase support. In another embodiment, the carrier is suitable for use in an industrial setting. Also provided is a non-physiological surface coated with an antibody or fragment thereof, as described herein. In one embodiment, the surface is in an industrial setting. In some embodiments, the fragment is an antigen-binding fragment.

Also provided are methods for producing antibodies, fragments, CDRs, or polypeptides, comprising or alternatively consisting of or further consisting of the step of culturing a host cell containing a polynucleotide encoding the antibody, its antigen-binding fragment, polypeptide, or CDR, under conditions that allow expression of the polynucleotide, and optionally isolating the antibody, fragment, CDR, and/or polypeptide from the cell and/or cell culture. In one embodiment, the host cell is a mammalian cell.

In one aspect, a method is provided for inhibiting or competing for the binding of a DNABII polypeptide or protein to microbial DNA. The method comprises or essentially consists of or additionally consists of contacting the DNABII polypeptide or protein with one or more antibodies, fragments thereof, a polypeptide, or a CDR, as disclosed herein. In one embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the head region of the IhfA5-mIhfB4NTHI chimeric peptide). In another embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the IhfA5-mIhfB4NTHI chimeric head peptide. In yet another embodiment, the contact is carried out in vivo or in vitro.

In another aspect, a method for disrupting a biofilm is provided. The method comprises or essentially consists of or additionally consists of contacting the biofilm with one or more antibodies, fragments thereof, a polypeptide, or a CDR, as disclosed herein. In one embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In another embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the IhfA5-mIhfB4NTHI chimeric head peptide. In yet another embodiment, the contact is made in vivo or in vitro.

Also further provided are methods for preventing the formation or destruction of a biofilm on a surface, comprising or alternatively consisting of or further consisting of treating a surface sensitive to or containing a biofilm, for example, with one or more antibodies, antigen-binding fragments thereof, a polypeptide or CDR, as described herein, wherein the antibody, fragment thereof, polypeptide or CDR binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide). In another embodiment, the antibody, fragment thereof, polypeptide or CDR binds the IhfA5-mIhfB4NTHI chimeric head peptide. In yet another embodiment, the treatment is carried out in vivo or in vitro.

Also further provided are methods for detecting a biofilm on a surface, comprising or alternatively consisting of or further consisting of contacting a surface (in one aspect susceptible to a biofilm or containing a biofilm) with one or more antibodies, antigen-binding fragments thereof, polypeptide or CDR, as described herein, wherein the antibody, fragment thereof, polypeptide or CDR binds to the tail or head region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, the antibody, fragment thereof, polypeptide or CDR binds the IhfA5-mIhfB4NTHI chimeric head peptide. In one embodiment, the contact is made in vivo or in vitro.

In one aspect, a method for detecting a microbial infection that leads to biofilm formation in a subject is provided. The method comprises or alternatively consists of, or further consists of, contacting one or more antibodies, fragments thereof, a polypeptide, or a CDR, as disclosed herein, with a biological sample suspected of containing a biofilm and isolated from the subject's body, and detecting binding of the antibody, fragment thereof, polypeptide, or CDR to any biofilm in the sample. The amount can be determined by the attending physician or veterinarian, for example, an effective amount for the subject. In one embodiment, the antibody, fragment, polypeptide, or CDR thereof binds to the tail or head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, the contact is made in vivo or in vitro.

In another aspect, a method is provided for screening subjects having a biofilm, comprising or alternatively consisting of, or further consisting of, contacting with one or more antibodies, fragments thereof, polypeptide, or CDR, as disclosed herein, a biological sample containing a biofilm and isolated from the subject, and determining the binding of the antibody, fragment thereof, polypeptide, or CDR to any biofilm in the sample. In one embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the tail or head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, a subject in whom binding is detected is selected for administration of one or more antibodies, fragments thereof, polypeptides, or CDRs as disclosed herein, and/or one or more polynucleotides or vectors encoding an antibody, fragment thereof, polypeptide, or CDR, wherein the antibody, fragment thereof, polypeptide, or CDR binds to the head region of the DNABII peptide (including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). The amount can be determined by the attending physician or veterinarian, for example, an effective amount for the subject. In another embodiment, the contact is carried out in vivo or in vitro.

The present application provides methods for detecting a biofilm in a subject by administering to the subject one or more antibodies or fragments thereof, as disclosed herein, and detecting any binding of the antibody, fragment thereof, polypeptide, or CDR, as disclosed herein, to the biofilm. In one aspect, the antibody, fragment thereof, polypeptide, or CDR binds to the tail or head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, the method further comprises detecting binding of the antibody, fragment thereof, polypeptide, or CDR to the biofilm.

In one aspect, methods are provided for preventing or disrupting a biofilm in a subject, comprising or alternatively consisting of or further consisting of administering to the subject one or more antibodies, fragments thereof, a polypeptide, or CDR as described herein, and/or one or more polynucleotides or vectors encoding the antibody, fragment thereof, polypeptide, or CDR, wherein they bind to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). The amount can be determined by the attending physician or veterinarian, for example, an effective amount for the subject. In one embodiment, the method further comprises detecting a biofilm by contacting one or more antibodies, fragments thereof, polypeptides or CDRs as disclosed herein with a sample suspected of containing a biofilm and detecting binding of the biofilm and the antibody, fragment thereof, polypeptide or CDR, and wherein the antibody, fragment thereof, polypeptide or CDR binds to the head region or tail region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide).

In another aspect, methods are provided for inhibiting, preventing, or treating a microbial infection that causes biofilm formation in a subject, comprising or alternatively consisting of or further consisting of administering to the subject one or more antibodies, fragments thereof, a polypeptide, or CDR, as described herein, and/or one or more polynucleotides or vectors encoding the antibody, fragment thereof, polypeptide, or CDR, wherein they bind to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). The amount can be determined by the attending physician or veterinarian, for example, an effective amount for the subject. In one embodiment, the method further comprises detecting a biofilm by contacting one or more antibodies, fragments thereof, polypeptides or CDRs as disclosed herein with a sample suspected of containing a biofilm and detecting binding of the biofilm and the antibody, fragment thereof, polypeptide or CDR, and wherein the antibody, fragment thereof, polypeptide or CDR binds to the head region or tail region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide).

In another aspect, methods for preventing or treating a condition characterized by biofilm formation in a subject are provided by administering to the subject one or more antibodies, fragments thereof, a polypeptide, or CDR as disclosed herein, and/or one or more polynucleotides or vectors encoding the antibody, fragment thereof, polypeptide, or CDR, wherein they bind to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric peptide). The amount can be determined by the attending physician or veterinarian, for example, an effective amount for the subject. Non-limiting examples of conditions include chronic non-healing wounds, lung infections caused by Burkholderia sp., venous ulcers, diabetic foot ulcers, ear infections, sinus infections, urinary tract infections, gastrointestinal diseases, hospital-acquired pneumonia, ventilator-associated pneumonia, surgical implant-associated infections, pulmonary infections, respiratory tract infections, cystic fibrosis, chronic obstructive pulmonary disease, catheter-associated infections, implanted device-associated infections, implanted prosthetic device-associated infections, osteomyelitis, cellulitis, abscesses, and periodontal disease. In one embodiment, the method further comprises detecting a biofilm by contacting one or more antibodies, fragments thereof, a polypeptide or CDR as disclosed herein with a sample suspected of containing a biofilm and detecting binding of the biofilm and the antibody, fragment thereof, polypeptide or CDR, and wherein the antibody, fragment thereof, polypeptide or CDR binds to the head region or tail region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide).

In some embodiments of the method disclosed herein, administering one or more antibodies, fragments thereof, a polypeptide, or a CDR reduces the level of one or more proinflammatory cytokines in a subject. Non-limiting examples of proinflammatory cytokines include: IL-1β, IL6, IL8, IL12p70, IL17A, interferon (IFN), and tumor necrosis factor (TNF). Additionally or alternatively, administering one or more antibodies, fragments thereof, a polypeptide, or a CDR increases the level of one or more anti-inflammatory cytokines in the subject. In one embodiment, anti-inflammatory cytokines include, but are not limited to, IL-10, IL-13, IL-1ra, IL-4, IL-11, and transforming growth factor-β (TGF-β).

Also provided is a method for generating passive immunity in a subject, comprising or alternatively consisting essentially of or additionally consisting of administering to the subject one or more antibodies, fragments thereof, a polypeptide or CDR, as disclosed herein, and/or one or more polynucleotides or vectors encoding the antibody, fragment, polypeptide or CDR thereof, wherein the antibody, fragment, polypeptide or CDR binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region and/or the IhfA5-mIhfB4NTHI chimeric head peptide). The amount can be determined by the attending physician or veterinarian, for example, an effective amount for the subject.

In one aspect, therapeutic methods are combined with diagnostic methods for detecting and/or monitoring biofilm formation and destruction using an antibody, fragment, polypeptide, or CDR thereof as disclosed herein.

In some embodiments, the biofilm is formed (i.e., produced) by Gram-negative or Gram-positive biofilm-producing bacteria. In some embodiments, the biofilm comprises the DNABII protein. In another embodiment, the biofilm comprises a histone-like protein from E. coli U93 (HU) or an integration host factor (IHF) binding protein. In some embodiments, the DNABII peptide is an IHF peptide. Additionally or alternatively, the DNABII peptide is an HU peptide. In some embodiments, the head region of the DNABII peptide is an IHFA head region and/or an IHFB head region. In another embodiment, the head region of the DNABII peptide is an IHFA head region conjugated directly or indirectly (e.g., via a linker) to an IHFB head region. In another embodiment, the head region of the DNABII peptide is the IhfA5-mIhfB4NTHI chimeric head peptide. In some embodiments, the tail region of the DNABII peptide is the IHFA tail region and/or the IHFB tail region. In another embodiment, the tail region of the DNABII peptide is the IHFA tail region conjugated directly or indirectly (e.g., via a linker) to the IHFB tail region. In yet another embodiment, the tail region of the DNABII peptide is the IhfA3-IhfB2NTHI chimeric tail peptide.

Also further provided are methods for producing an interfering nucleic acid or alternatively consisting of or further consisting of producing a nucleic acid consisting of about 10-20 nucleotides that specifically binds to a specific binding partner of an antibody or fragment thereof as disclosed herein, and optionally isolating the interfering nucleic acid obtained by this method.

In another aspect, the present application provides a non-physiological surface coated with one or more compounds selected from an antibody, fragment thereof, polypeptide, or CDR as disclosed herein, and optionally, in one aspect, the surface is in an industrial setting.

Methods for producing an antiserum effective for disrupting a biofilm are also provided, comprising immunizing a subject with a small molecule and obtaining the antiserum from the subject and, optionally, isolating polyclonal antiserum or monoclonal antibodies from the subject.

Kits are also provided. The kits comprise one or more antibodies, fragments thereof, a polypeptide or CDR, a polynucleotide, a vector, a host cell, or a composition as described herein, and, optionally, instructions for use.

Brief description of the drawings

Figure 1 is a schematic showing how the DNABII protein (e.g., IHF) binds to double-stranded DNA, as well as the two arms of the DNABII protein.

Figures 2A–2D show the results of a direct binding ELISA performed to assess the binding of humanized monoclonal antibody variants to their respective peptides. The head or tail chimeric peptide was added to the wells of a 96-well plate at a concentration of 2 μg/mL, and an 8-point dilution series of antibodies (starting concentration 50 μg/mL, dilution 1:3) was added. HRP-conjugated anti-human IgG Fc antibody (1:7000, Jackson ImmunoResearch Laboratories, 109-035-098) was used as a secondary detection antibody. A standard direct ELISA protocol was followed, and optical density was measured at 450 nm using a microplate reader. _EC values were calculated, and the humanized variants were determined to have comparable binding. Humanized antibodies to the head chimeric peptide are shown in Figures 2A and 2B. Humanized antibodies to the tail chimeric peptide are shown in Figures 2C and 2D.

Figure 3 is an in vitro model for reversing established biofilm in an 8-well slide using a humanized monoclonal antibody to the head or tail chimeric peptide as described herein.

Figures 4A-4G present a comparison (in table form, Figure 4A and detailed alignments, Figures 4B-4G) of the amino acid identities of the heavy chains and light chains of humanized monoclonal antibodies designed to target the IhfA5-mIhfB4NTHI chimeric head peptide. Amino acid alignments were performed using NCBI blastp.

Figures 5A-5G present a comparison (in table form, Figure 5A and detailed alignments, Figures 5B-5G) of the amino acid identities of the heavy chains and light chains of humanized monoclonal antibodies designed to target the IhfA3-IhfB2NTHI chimeric head peptide. Amino acid alignments were performed using NCBI blastp.

Figure 6 shows relative biofilm disruption data using humanized monoclonal antibodies designed to target the IhfA5-mIhfB4NTHI head peptide. Biofilm disruption: NTHI 86-028NP colonies were picked from overnight chocolate agar cultures and suspended in brain heart infusion broth supplemented with 2 μg β-NAD and heme per 1 ml of medium (sBHI). The optical density at 490 nm was then adjusted to 0.65, and the culture was diluted 1:6 in sBHI before static incubation at 37°C with 5% _CO2 for 3 h. The culture was then diluted 1:2500 in fresh sBHI, and 200 μl aliquots of the suspension were added to each well of an 8-well microscope slide. The slide was then statically incubated at 37°C with 5% _CO2 for 3 h. After 16 h, 200 μl of fresh sBHI was added to each well, and the slide was incubated for an additional 8 h. At this time, the medium was aspirated from each well, and 5 μg of monoclonal antibody per well was added. The biofilms were incubated for an additional 16 h. The biofilms were then washed and stained with FM1-43FX bacterial cell membrane dye (Invitrogen) and fixed overnight at 4°C in 16% paraformaldehyde, 2.5% glutaraldehyde, 4.0% acetic acid in 0.1 M phosphate buffer (pH 7.4). The fixative solution was removed by aspiration, and 200 μl of Hanks' balanced salt solution was added to each well before viewing the biofilms on a Zeiss 800 Meta-laser scanning confocal microscope. Images were compiled using Zeiss Zen Black software, and biofilm biomass was calculated using COMSTAT2.1 software. The K _A for the IhfA5-mIhfB4NTHI head chimeric peptide (in 1/M) ranges from approximately 4E+05 to approximately 2E+08.

Figure 7 shows relative biofilm disruption data using a humanized monoclonal antibody designed to target the IhfA3-IhfB2NTHI tail chimeric peptide. Biofilm disruption: NTHI 86-028NP colonies were picked from an overnight culture on chocolate agar and suspended in brain heart infusion broth supplemented with 2 μg β-NAD and heme per 1 ml of medium (sBHI). The optical density at 490 nm was then adjusted to 0.65, and the culture was diluted 1:6 in sBHI before static incubation at 37°C with 5% _CO2 for 3 h. The culture was then diluted 1:2500 in fresh sBHI, and 200 μl aliquots of the suspension were added to each well of an 8-well microscope slide. The slide was then statically incubated at 37°C with 5% _CO2 for 3 h. After 16 h, 200 μl of fresh sBHI was added to each well, and the slide was incubated for an additional 8 h. At this time, the medium was aspirated from each well, and 5 μg of monoclonal antibody per well was added. The biofilms were incubated for an additional 16 h. The biofilms were then washed and stained with FM1-43FX bacterial cell membrane dye (Invitrogen) and fixed overnight at 4°C in 16% paraformaldehyde, 2.5% glutaraldehyde, 4.0% acetic acid in 0.1 M phosphate buffer (pH 7.4). The fixative solution was removed by aspiration, and 200 μl of Hanks' balanced salt solution was added to each well before viewing the biofilms on a Zeiss 800 Meta-laser scanning confocal microscope. Images were compiled using Zeiss Zen Black software, and biofilm biomass was calculated using COMSTAT2.1 software. The K _A for the IhfA3-IhfB2NTHI tail chimeric peptide (in 1/M) ranges from approximately 8E+06 to approximately 2E+09.

Figures 8A–8B show that murine monoclonal antibody Fab fragments directed against the β-head portion disrupt biofilms formed by five human pathogens in vitro. FIGURE 8A shows representative images of bacterial biofilms (pseudo-stained white) demonstrating significant biofilm disruption by 170 nM intact IgG or Fab directed against the β-head portion. Orthogonal projections show a top-down view to depict the spatial distribution of the biofilm in the x-y plane, and the side view indicates the biofilm height in the z-plane (arrows). Scale bars, 20 μm. FIGURE 8B shows the biomass in each image quantified using COMSTAT2 software. Each assay was repeated three times on different days, and the mean ± SEM was determined. *P≤0.05, **P≤0.01 compared with the corresponding intact Fabs or IgG (one-way ANOVA with multiple comparisons).

Figures 9A–9D illustrate the killing of NTHI (non-encapsulated H. influenzae bacteria) resident in biofilms and the resorption of middle ear mucosal biofilms mediated by Fab fragments of a mouse monoclonal antibody against the β-head portion. Figure 9A shows the timeline and treatment assignment. A dose of 342 nM Fab fragment was administered to each middle ear. FIGURE 9B shows the relative amount of NTHI in mucosal biofilm and attached to middle ear mucosa among treatment groups. Figure 9C shows the relative mucosal biofilm quantification, as determined by six reviewers blinded to treatment, among treatment groups. Figures 9B and 9C: 6–8 middle ears were examined per group, individual ear values and the group mean are shown, and the groups for each panel from left to right represent isotype controls with Fab, anti-β-tail Fab, and anti-β-head Fab. *P≤0.05, **P≤0.01, ***P≤0.001 (one-way ANOVA with multiple comparisons). Figure 9D shows representative images of middle ears of chinchillas from each group, the mean biofilm score in the ear mucosa is indicated in the box in the lower right corner. TM, tympanic membrane; S, bony septa; MEM, middle ear mucosa; B, biofilm (outlined). In one embodiment, delivery of Fab fragments of mouse monoclonal antibodies against the β-head portion of IHF both significantly reduced the bacterial load in the middle ear and destroyed mucosal biofilms, however, Fab fragments against the β-tail portion not only failed to achieve this result, but were also associated with significant inflammation.

Figures 10A-10B show proinflammatory cytokines predominant in middle ear fluids from animals treated with 342 nM Fab fragments of mouse monoclonal antibodies against the β-tail. FIGURE 10A shows the relative amounts of proinflammatory cytokines in middle ear fluids of chinchillas after NTHI challenge and Fab fragment treatment, as determined by bead cytometric analysis. *P≤0.05, **P≤0.01 compared with Fab fragments against the β-head (one-way ANOVA with multiple comparisons), ǂ P≤0.05 compared with isotype control Fab fragments (unpaired t-test for independent samples). FIGURE 10B shows the relative mean concentration of the anti-inflammatory cytokine IL-10 among the three Fab-treated groups. *P≤0.05 versus anti-β-tail Fab, ++P≤0.01 versus anti-β-tail Fab, or isotype control Fab (one-way ANOVA with multiple comparisons). Five to seven middle ear fluids were tested per group. Mean cytokine concentrations ± standard deviations are shown. For each panel (FIGURE 10A or 10B), cytokine results are shown from left to right for the isotype control Fab, anti-β-tail Fab, and anti-β-head Fab groups. In one embodiment, delivery of Fab fragments against the β-head portion resulted in a significant reduction in the concentration of six pro-inflammatory cytokines and a significantly greater concentration of the anti-inflammatory cytokine IL-10, which supports relative inflammation or its absence, as shown in Figure 9D.

Figures 11A–11D show that anti-chimeric head-band Fab fragments from polyclonal rabbit IgG mediated the clearance of resident NTHI in biofilm, eradication of established mucosal biofilms, and normalization of experimental disease. Figure 11A shows the timeline of the study. A dose of 342 nM Fab was administered to each middle ear. Figure 11B shows the relative amount of NTHI present in mucosal biofilms and adhered to middle ear mucosa one and seven days after completion of antibody therapy. Figure 11C shows the relative amount of remaining mucosal biofilm as determined by six reviewers blinded to treatment. Figures 11B and 11C: Six middle ears per group, values for individual ears, and the mean value for each group shown. Groups for each graph from left to right represent naive serum Fabs, anti-tail chimeric Fabs, and anti-head chimeric Fabs. *P≤0.05, **P≤0.01, ***P≤0.001 (one-way ANOVA with multiple comparisons). Figure 11D shows representative images of middle ear mucosal biofilms; mean mucosal biofilm score is indicated in the box in the lower right corner. TM, tympanic membrane; S, bony septa; MEM, middle ear mucosa; B, biofilm (circled). Rabbit polyclonal antibody Fab fragments directed against the head chimeric peptide mediated rapid and robust biofilm clearance of resident NTHI, eradication of established mucosal biofilms, and normalization of experimental OM. Fab fragments of rabbit polyclonal antibodies directed against the tail chimeric peptide did not induce biofilm clearance of resident NTHI or disease normalization, but were instead associated with significant inflammation (Figure 11D), as were Fab fragments of mouse monoclonal antibodies directed against the β-tail region (see Figure 9D).

Figures 12A-12B show that the humanized monoclonal antibody against the head chimeric peptide disrupted bacterial biofilms in vitro. Figure 12A shows representative images of bacterial biofilms (pseudo-stained white) after 16-hour incubation with HuTipMab or HuTailMab. Orthogonal projections show a top-down view to depict the spatial distribution of the biofilm in the x-y plane, and the side view indicates the biofilm height in the z-plane (arrows). Scale bars, 20 μm. As used herein, the designation "HuTipMab" refers to a monoclonal antibody (Mab or mAb) comprising the anti-head HC1 domain having the amino acid sequence of SEQ ID NO: 1 and the anti-head LC1 domain having the amino acid sequence of SEQ ID NO: 7. FIGURE 12B shows the percentage of biofilm biomass remaining after exposure to HuTipMab compared to HuTailMab, determined using the COMSTAT2 assay. Mean ± standard deviation is shown. Experiments were performed three times on different days. ***P ≤ 0.001 (unpaired t-test). The HuTipMab antibody significantly disrupted biofilms formed by various respiratory pathogens in vitro. As used herein, the term "HuTailMab" refers to a monoclonal antibody (Mab or mAb) comprising an anti-tail antibody HC1 domain having the amino acid sequence of SEQ ID NO: 4 and an anti-tail antibody LC1 domain having the amino acid sequence of SEQ ID NO: 10.

Figures 13A–13D show that the humanized anti-tail chimeric peptide monoclonal antibody (HuTipMab) eliminated preexisting NTHI biofilms in the middle ear during experimental NTHI-induced OM. Figure 13A shows the study and treatment timeline. Figure 13B shows the relative amount of NTHI in the middle ear. Figure 13C shows the relative amount of NTHI mucosal biofilm remaining in the middle ear after treatment with the human monoclonal antibodies. Figures 13B and 13C: Six middle ears per group, showing individual ear values and the mean value for the group. The groups for each graph, from left to right, represent the groups treated with saline, the humanized anti-tail chimeric peptide monoclonal antibody, and the humanized anti-head chimeric peptide monoclonal antibody. ****P ≤ 0.0001 (one-way ANOVA with multiple comparisons). Figure 13D shows representative images of the middle ear from each group; the mean mucosal biofilm score for the ear is indicated in the box in the lower right corner. S, bony septa; MEM, middle ear mucosa; B, biofilm, circled. The HuTipMab antibody eliminated NTHI by dissolving mucosal biofilms, which was a rapid and robust result. It is also noteworthy that, when humanized, the monoclonal antibody targeting the tail chimeric peptide is no longer associated with significant inflammatory changes, as was consistently observed with either the mouse monoclonal antibody targeting the β-tail region (see Figure 9D) or the Fab fragments of rabbit polyclonal antibodies targeting the tail chimeric peptide (see Figure 11D).

Figures 14A-14B show that proinflammatory cytokines were significantly less abundant in the middle ear fluids of animals treated with HuTipMab. Relative amounts of proinflammatory and anti-inflammatory cytokines in the middle ear fluids of chinchillas after NTHI challenge and treatment with humanized monoclonal antibodies (Figure 14A) six days after NTHI challenge (one day after completion of therapy) and (Figure 14B) 13 days after NTHI challenge (seven days after completion of therapy), as determined by bead cytometric analysis. *P≤0.05 vs. saline, **P≤0.01 vs. saline, ***P≤0.001 vs. saline, ****P≤0.0001 vs. saline, +P≤0.05 vs. HuTailMab, ++P≤0.01 vs. HuTailMab, ++P≤0.001 vs. HuTailMab, ++++P≤0.0001 vs. HuTailMab (one-way ANOVA with multiple comparisons). Three to five middle ear fluids were tested per group. Mean cytokine concentrations ± SD are shown. Pro-inflammatory cytokine levels in animals treated with HuTailMab were now equivalent to those in animals treated with saline, confirming the absence of a link between the administration of this now humanized monoclonal antibody compared to mouse monoclonal antibodies.

Figures 15A–15D show immunization with the head chimeric peptide preventing ascending experimental OM in a viral-bacterial coinfection model. Figure 15A shows the study timeline and the vaccine formulations delivered. Figure 15B shows the relative amounts of NTHI in nasopharyngeal lavage fluids one day after bacterial challenge to ensure that all groups were equally colonized. Eight lavage fluids per group, NS, not significant, one-way ANOVA. Figure 15C shows the number of animals in each group with evidence of experimental OM, i.e., inflammation and/or middle ear fluid visualized by blinded video otoscopy. Eight animals per group, ****P ≤ 0.001 compared with dmLT or tail chimeric peptide, Mantel-Cox test. Figure 15D shows representative images of the tympanic membranes of each group on day 11, which was the day of peak disease incidence in the group immunized with the head-on chimeric peptide. TM, tympanic membrane. Immunization with the head-on chimeric peptide prevented ascending polymicrobial OM and promoted rapid resolution of the observed limited disease.

Detailed description

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this disclosure pertains. All nucleotide sequences presented herein are in the 5' to 3' direction. Although any methods and materials similar or equivalent to those disclosed herein can be used in the practice or testing of the present invention, specific, non-limiting, exemplary methods, devices, and materials are described herein. All technical and patent publications cited herein are hereby incorporated by reference in their entirety. Nothing in this description should be construed as an admission that the present invention does not precede the disclosure in these publications.

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of tissue culture, immunology, molecular biology, microbiology, cell biology, and recombinant DNA, which are within the skill of the art. See, e.g., Sambrook and Russell eds. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition; Ausubel et al. eds. (2007) Current Protocols in Molecular Biology series; Methods in Enzymology series (Academic Press, Inc., N.Y.); MacPherson et al. (1991) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual; Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition; Gait ed. (1984) Oligonucleotide Synthesis; Pat. US No. 4683195; Hames and Higgins eds. (1984) Nucleic Acid Hybridization; Anderson (1999) Nucleic Acid Hybridization; Hames and Higgins eds. (1984) Transcription and Translation; Immobilized Cells and Enzymes (IRL Press (1986)); Perbal (1984) A Practical Guide to Molecular Cloning; Miller and Calos eds, (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); and Herzenberg et al. eds (1996) Weir's Handbook of Experimental Immunology.

All numerical designations, such as pH, temperature, time, concentration, and molecular weight, including ranges, are approximate values that vary (+) or (-) in increments of 1.0 or 0.1, as appropriate, or alternatively by changing by +/- 15%, or alternatively 10%, or alternatively 5%, or alternatively 2%. It should be understood, although not always explicitly stated, that all numerical designations are preceded by the term "about". It should also be understood, although not always explicitly stated, that the reagents described herein are merely illustrative and that their equivalents are known in the art.

The singular forms "a" and "an" used in the description and claims should be construed as encompassing both singular and plural meanings unless the context clearly dictates otherwise. For example, the term "polypeptide" encompasses a plurality of polypeptides, including mixtures thereof.

As used herein, the term "comprising" is intended to indicate that the compositions and methods include the listed elements, but do not exclude the presence of others. The term "consisting essentially of," when used to define compositions and methods, means to exclude other elements that are of any significant importance to the combination for the purpose of the intended use. Thus, a composition consisting essentially of the elements as defined herein does not exclude the presence of trace impurities due to the isolation and purification process, and pharmaceutically acceptable carriers such as phosphate-buffered saline, preservatives, and the like. The term "consisting of" is understood to exclude more than trace elements of other ingredients and essential steps of the method of administration disclosed herein. Embodiments defined by each of these transitional terms are included within the scope of the present invention.

The term "biofilm" refers to an organized community of microorganisms that intermittently adhere to the surface of a structure, which may be organic or inorganic, along with polymers such as DNA that they secrete, release, and/or become accessible to the extracellular environment through bacterial lysis. Biofilms are highly resistant to microbiotics and antimicrobial agents. They live on gingival tissue, teeth, and fillings, causing dental caries and periodontal disease, also known as periodontal plaque. They also cause chronic middle ear infections. Biofilms can also form on the surface of dental implants, stents, catheter lines, and contact lenses. They grow on pacemakers, heart valve replacements, artificial joints, and other surgical implants. The Centers for Disease Control estimates that more than 65% of nosocomial (hospital-acquired) infections are caused by biofilms. They cause chronic vaginal infections and lead to life-threatening systemic infections in people with weakened immune systems. Biofilms are also associated with numerous diseases. For example, patients with cystic fibrosis have Pseudomonas infections, which often lead to the formation of antibiotic-resistant biofilms. In one embodiment, the biofilm comprises the DNABII polypeptide or protein. In another embodiment, the biofilm comprises IHF and/or HU. In yet another embodiment, the biofilm comprises IHFA and/or IHFB.

The term "disruption" refers to a reduction in the formation of the DNA/protein matrix, which is a component of the microbial biofilm. In some embodiments, biofilm disruption refers to biofilm dispersal, the release of microorganisms from the DNA/protein matrix of the biofilm, and optional killing of the microorganisms by host immune effectors and/or antibiotics.

"DNABII polypeptide or protein" means a DNA-binding protein or polypeptide that consists of DNA-binding domains and thus has a specific or general affinity for microbial DNA. In one aspect, these domains bind DNA in the minor groove. Non-limiting examples of DNABII proteins include the integration host factor (IHF) protein and the histone-like protein from E. coli strain U93 (HU). Other DNA-binding proteins that may be associated with biofilm include DPS (Genbank accession number: CAA49169), H-NS (Genbank accession number: CAA47740), Hfq (Genbank accession number: ACE63256), CbpA (Genbank accession number: BAA03950), and CbpB (Genbank accession number: NP-418813).

The "integration host factor" or "IHF" protein is a bacterial protein used by bacteriophages to incorporate their DNA into host bacteria. Bacteriophages also bind extracellular microbial DNA. The genes encoding the IHF protein subunits in E. coli are himA (Genbank accession number: POA6X7.1) and himD (POA6Y1.1). Homologs of these genes are found in other organisms. In some embodiments, the term "IHF" refers to one or both of the two IHF subunits: the integration host factor alpha subunit (IHFA or IhfA) and the integration host factor beta subunit (IHFB or IhfB).

"HU" or "histone-like protein from E. coli strain U93" refers to a class of heterodimeric proteins commonly associated with E. coli. HU proteins are known to bind to DNA structures. Related proteins have been isolated from other microorganisms. The complete amino acid sequence of E. coli HU was determined by Laine et al. (1980) Eur. J. Biochem 103(3)447–481. Antibodies to the HU protein are commercially available from Abeam. The genes encoding the HU protein subunits from E. coli are hupA and hupB, corresponding to SEQ ID NOs: 29 and 30, in that order. Homologs for these genes have been found in other organisms, and peptides corresponding to these genes from other organisms can be found in Table 10 of WO 2011/123396.

The term "surface antigens" or "surface proteins" refers to proteins or peptides on the surface of cells, such as bacterial cells. Examples of outer membrane proteins are, for example, OMP P5 (Genbank accession number: YP-004139079.1), OMP P2 (Genbank accession number: ZZX87199.1), and OMP P26 (Genbank accession number: YP-665091.1), while examples of surface antigens are rsPilA or recombinant soluble PilA (Genbank accession number: EFU96734.1) and pilin type IV (Genbank accession number: Yp-003864351.1).

The name "Haemophilus influenzae" refers to pathogenic bacteria that can cause a variety of infections, including ear infections, eye infections, and sinusitis. Many different strains of Haemophilus influenzae have been isolated, each carrying the IhfA, ihfB, and hupA genes or proteins. Some non-limiting examples of different Haemophilus influenzae strains include Rd KW20, 86-028NP, R2866, PittGG, PittEE, R2846, and 2019.

"Microbial DNA" refers to single-stranded or double-stranded DNA from a microorganism that is embedded in a biofilm.

"Inhibition, prevention, or disruption" of biofilm refers to the prophylactic or therapeutic reduction of biofilm structure.

The term "bent polynucleotide" refers to a double-stranded polynucleotide that contains a small loop on one strand that does not pair with the other strand. In some embodiments, the loop is from 1 to about 20 bases long, or alternatively from 2 bases to about 15 bases long, or alternatively from about 3 bases to about 12 bases long, or alternatively from about 4 bases to about 10 bases long, or alternatively has about 4, 5, 6, 7, 8, 9, or 10 bases.

"Polypeptides that compete for DNA binding with DNABII proteins, such as IHF," refers to proteins or peptides that compete with DNABII (e.g., IHF) for binding to bent or distorted DNA structures, but do not form a biofilm with DNA. Examples, but not limited to, include fragments of IHF that contain one or more IHF DNA-binding domains, or their biological equivalents.

The "object (subject)" of diagnosis or treatment is a cell or an animal, such as a mammal, including humans. Non-human animals to be diagnosed or treated, as well as subjects to be infected or used in animal models, include, for example, monkeys, murine animals such as rats, mice, chinchillas, canines such as dogs, leporids such as rabbits, as well as livestock, sport animals, and companion animals. The terms "object (subject)," "host," "individual," and "patient" are used interchangeably herein to refer to animals, typically mammals. Non-limiting examples of mammals include humans, primates (e.g., great apes, gibbons, chimpanzees, orangutans, marmosets, macaques, etc.), domestic animals (e.g., dogs and cats), farm animals (e.g., horses, cows, goats, sheep, pigs), and experimental animals (e.g., mice, rats, rabbits, guinea pigs). In some embodiments, the mammal is a human. The mammal may be of any age or at any stage of development (e.g., an adult, an adolescent, a child, an infant, or a mammal in utero). The mammal may be male or female. In some embodiments, the subject is a human.

The terms "protein," "peptide," and "polypeptide" are used interchangeably and in the broadest sense to refer to a compound consisting of two or more subunit amino acids, amino acid analogs, or peptidomimetics. The subunits may be linked by peptide bonds. In another embodiment, the subunits may be linked by other bonds, such as ester bonds, ether bonds, etc. A protein or peptide must contain at least two amino acids, and there is no limitation on the maximum number of amino acids that can make up a protein or peptide sequence. As used herein, the term "amino acid" refers to natural and/or unnatural or synthetic amino acids, including glycine and the D and L optical isomers, amino acid analogs, and peptidomimetics.

"C-terminal polypeptide" means at least 10, or alternatively at least 15, or alternatively at least 20, or at least 25 C-terminal amino acids, or alternatively, half of the polypeptide. In another aspect, for polypeptides containing 90 amino acids, the C-terminal polypeptide will contain amino acids 46 to 90. In one aspect, the term means 20 C-terminal amino acids from the carboxyl terminus.

A "head fragment" of a DNABII polypeptide means a DNABII polypeptide that, with the help of, for example, IHF-alpha and IHF-beta, forms two protein arms (see FIGURE 1). Non-limiting examples include IhfA, head fragment A: NFELRDKSSRPGRNPKTGDVV, SEQ ID NO: 31, and IhfB, head fragment B: SLHHRQPRLGRNPKTGDSVNL, SEQ ID NO: 32.

The "tail fragment" of a DNABII polypeptide means the region of the protein that is both exposed to the general environment and not occluded by DNA or other polypeptides.

An immunodominant antigen refers to a region of a protein that is recognized and bound by an antibody with high affinity.

An immunoprotective antigen refers to a region of a protein that, when recognized and bound with high affinity by an antibody, interferes with the protein's function; antibodies generated against an immunoprotective antigen are characterized by enhanced or optimal activity against the target as a result of interfering with the protein's function—in this case, improved biofilm clearance.

The terms "polynucleotide" and "oligonucleotide" are used interchangeably and refer to the polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or their analogs. Polynucleotides may have any three-dimensional structure and may perform any function, known or unknown. The following are non-limiting examples of polynucleotides: a gene or gene fragment (e.g., a probe, primer, EST or SAGE tag), exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, interfering RNA (iRNA), ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. A polynucleotide may contain modified nucleotides, such as methylated nucleotides and nucleotide analogs. If modified nucleotides are present, modifications to the nucleotide structure may be provided before or after assembly of the polynucleotide. The nucleotide sequence may be interrupted by non-nucleotide components. The polynucleotide may be further modified after polymerization, for example, by conjugation with a labeling component. The term also refers to both double-stranded and single-stranded molecules. Unless otherwise stated or required, any embodiment disclosed herein that represents a polynucleotide encompasses both the double-stranded form and each of the two complementary single-stranded forms known or suspected to constitute the double-stranded form.

A polynucleotide consists of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); thymine (T); and uracil (U) instead of thymine when the polynucleotide is RNA. Thus, the term "polynucleotide sequence" is an alphabetic representation of the polynucleotide molecule. This alphabetic representation can be entered into databases on a computer with a central processing unit and used for bioinformatics applications such as functional genomics and homology searching.

The term "isolated" or "recombinant" as used herein with respect to nucleic acids such as DNA or RNA refers to molecules separated from other DNA or RNA, respectively, that are also present in the natural source of the macromolecules, as well as from polypeptides. The term "isolated or recombinant nucleic acid" includes fragments of nucleic acid that do not occur in nature as fragments and cannot be found in the natural state. The term "isolated" is also used herein to refer to polynucleotides, polypeptides, and proteins that are separated from other cellular proteins, and the term is intended to encompass both purified and recombinant polypeptides. In other embodiments, the term "isolated or recombinant" refers to compounds and structures separated from cellular and other constituents with which the cell, tissue, polynucleotide, peptide, polypeptide, protein, antibody, or fragment(s) thereof are normally associated in nature. For example, an isolated cell is one that is separated from tissue or cells with a dissimilar phenotype or genotype. An isolated polynucleotide is separated from the 3' and 5' contiguous nucleotides to which the polynucleotide is normally associated in its native or natural environment, such as a chromosome. As will be appreciated by those skilled in the art, a non-naturally occurring polynucleotide, peptide, polypeptide, protein, antibody, or fragment(s) thereof does not require "isolation" to distinguish it from its naturally occurring counterparts.

It should be understood without explicit recitation and unless otherwise implied that when the present invention relates to a polypeptide, protein, polynucleotide, or antibody, an equivalent or biological equivalent thereof is included within the scope of the present invention. As used herein, the term "biological equivalent" is synonymous with the term "equivalent" when referring to a reference protein, antibody, fragment, polypeptide, or nucleic acid, meaning those that have minimal homology while retaining the desired structure or functionality. Unless otherwise indicated herein, any polynucleotide, polypeptide, or protein referenced herein is also intended to include their equivalents. In one aspect, an equivalent polynucleotide is a polynucleotide that hybridizes under stringent conditions to a polynucleotide or complementary polynucleotide as described herein for use in the described methods. In another aspect, an equivalent antibody or antigen-binding polypeptide is an antibody that binds with at least 70%, or alternatively at least 75%, or alternatively at least 80%, or alternatively at least 85%, or alternatively at least 90%, or alternatively at least 95% with affinity or higher affinity to a reference antibody or antigen-binding fragment. In another aspect, equivalents compete for binding of the antibody or antigen-binding fragment to the corresponding antigen in a competitive ELISA assay. In another aspect, an equivalent means at least about 80% homology or identity, and alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 95%, or alternatively 98% homology or identity to a reference compound and exhibits a biological activity substantially equivalent to the activity of the reference protein, polypeptide, or nucleic acid. Examples of biologically equivalent polypeptides are shown in Table 9 of WO 2011/123396, which notes conservative amino acid substitutions of the disclosed amino acid sequences.

A polynucleotide or polynucleotide region (or polypeptide or polypeptide region) having a certain percentage (e.g., 80%, 85%, 90%, or 95%) of "sequence identity" with another sequence means that, when aligned, that percentage of bases (or amino acids) are the same when the two sequences are compared. Alignment and percentage of sequence homology or identity can be determined using programs known in the art, such as those described in Current Protocols in Molecular Biology (Ausubel et al., Eds. 1987) Supplement 30, section 7.7.18, Table 7.7.1. In some embodiments, default parameters are used for alignment. A non-limiting exemplary alignment program is BLAST using default parameters. Specifically, exemplary programs include BLASTN and BLASTP using the following default parameters: genetic code = standard; filter = none; strand = both; cutoff = 60; expectation = 10; matrix = BLOSUM62; descriptions = 50 sequences; sort by = HIGHEST SCORE; databases = non-redundant, GenBank+EMBL+DDBJ+PDB+GenBank CDS translations+SwissProtein+SPupdate+PIR. Detailed information on these programs can be found at the following web address: ncbi.nlm.nih.gov/cgi-bin/BLAST. Sequence identity and percent identity were determined by incorporating them into clustalW (available at align.genome.jp, last accessed March 7, 2011).

"Homology," or "identity," or "similarity" refers to the sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a specific position in each sequence, which can be aligned for comparison purposes. When a position in the compared sequence is occupied by the same base or amino acid, the molecules are homologous at that position. The degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. An "unrelated" or "non-homologous" sequence has less than 40% identity, or alternatively, less than 25% identity, with one of the sequences of the present invention.

"Homology," or "identity," or "similarity" can also refer to two nucleic acid molecules that hybridize under stringent conditions.

"Hybridization" refers to a reaction in which one or more polynucleotides react to form a complex stabilized by hydrogen bonds between the bases of the nucleotide residues. Hydrogen bonding can occur through Watson-Crick base pairing, Hoogsteen bonding, or any other sequence-specific mechanism. The complex may include two strands forming a duplex structure, three or more strands forming a multistranded complex, a single self-hybridizing strand, or any combination thereof. The hybridization reaction may represent a step in a larger process, such as the initiation of a PCR reaction or the enzymatic cleavage of a polynucleotide by a ribozyme.

Examples of stringent hybridization conditions include: incubation temperatures from about 25°C to about 37°C; hybridization buffer concentrations from about 6x SSC to about 10x SSC; formamide concentrations from about 0% to about 25%; and wash solutions from about 4x SSC to about 8x SSC. Examples of moderate hybridization conditions include: incubation temperatures from about 40°C to about 50°C; buffer concentrations from about 9x SSC to about 2x SSC; formamide concentrations from about 30% to about 50%; and wash solutions from about 5x SSC to about 2x SSC. Examples of highly stringent conditions include: incubation temperatures from about 55°C to about 68°C; buffer concentrations from about 1x SSC to about 0.1x SSC; formamide concentrations from about 55% to about 75%; and washing solutions of approximately 1× SSC, 0.1× SSC, or deionized water. Typically, hybridization incubation times range from 5 minutes to 24 hours, with 1, 2, or more washing steps, and wash incubation times of approximately 1, 2, or 15 minutes. SSC is 0.15 M NaCl and 15 mM citrate buffer. It is understood that equivalents of SSC using other buffer systems can be used.

As used herein, the term "expression" refers to the process by which polynucleotides are transcribed into mRNA and/or the process by which transcribed mRNA is subsequently translated into peptides, polypeptides, or proteins. If the polynucleotide is derived from genomic DNA, expression may involve mRNA splicing in a eukaryotic cell.

The term "encoding" when applied to polynucleotides refers to a polynucleotide known to "encode" a polypeptide if, in its native state or when manipulated using methods well known to those skilled in the art, the polynucleotide can be transcribed and/or translated to produce mRNA for the polypeptide and/or a fragment thereof. The antisense strand is the complement of such a polynucleotide, and the coding sequence can be deduced from it.

As used herein, the terms "treat," "treatment," and the like are used to mean obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing a disorder or a sign or symptom thereof, and/or may be therapeutic in terms of partially or completely curing the disorder and/or an adverse effect associated with the disorder. As used herein, the term "treat" or "treating" a disease in a subject may also refer to (1) preventing the onset of symptoms or a disease in a subject who is predisposed to or does not yet exhibit symptoms of the disease; (2) inhibiting the disease or stopping its progression; or (3) alleviating or regressing a disease or symptoms of a disease. As is understood in the art, "treating" is an approach to obtaining beneficial or desired results, including clinical results. For the purposes of the present invention, beneficial or desired results may include, but are not limited to, one or more results selected from the following: alleviation or improvement of one or more symptoms, reduction in the severity of a condition (including a disease), stabilization (i.e., not worsening) of a condition (including a disease), delay or slowing of the progression of a condition (including a disease), progression, improvement, or mitigation of a condition (including a disease), and remission (partial or complete), detectable or undetectable. In one aspect, treatment excludes prophylaxis.

The term "prevent" refers to the prevention (prophylaxis) of a disorder or effect in vitro or in vivo in a system or organism predisposed to the disorder or effect. An example is the prevention of biofilm formation in a system infected with a microorganism known to produce it.

A "composition" is intended to mean a combination of an active agent and another compound or composition, both inert (e.g., a detectable agent or label) and active, such as an adjuvant, diluent, binder, stabilizer, buffers, salts, lipophilic solvents, preservatives, adjuvants, etc., and includes pharmaceutically acceptable carriers. Carriers also include pharmaceutical excipients and additives, proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, di-, tri-, tetra-oligosaccharides, and oligosaccharides; derivatized sugars such as alditols, aldonic acids, esterified sugars, etc., and polysaccharides or polymers of sugars), which may constitute, individually or in combination, 1-99.99% of the composition by weight or volume. Typical protein excipients include serum albumin, such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Typical amino acid/antibody components that can also act as a buffering capacity include alanine, arginine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like. Carbohydrate excipients are also within the scope of the invention, examples of which include, but are not limited to, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, and the like; disaccharides such as lactose, sucrose, trehalose, cellobiose, etc.; polysaccharides such as raffinose, melezitose, maltodextrins, dextrans, starches, etc.; and alditols such as mannitol, xylitol, maltitol, lactitol, sorbitol (glucitol), and myoinositol.

A "pharmaceutical composition" is intended to include a combination of an active agent with a carrier, inert or active, which makes the composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

"Pharmaceutically acceptable carriers" refer to any diluents, excipients, or delivery agents that can be used in the compositions disclosed herein. Pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering agents such as phosphates, glycine, sorbic acid, potassium sorbate, partial glycerides, mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and wool fat. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, the standard reference work in this field. They can be selected according to the intended form of administration, i.e., oral tablets, capsules, elixirs, syrups, etc., and are consistent with standard pharmaceutical practice.

The compositions used in accordance with the invention can be packaged in dosage form for ease of administration and uniformity of dosage. The term "unit dose" or "dosage" refers to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of the composition calculated to elicit the desired responses in connection with its administration, i.e., appropriate to the route of administration and treatment regimen. The amount administered, which corresponds to the number of treatment procedures and the dosage, will depend on the desired result and/or protective effect. The exact amounts of the composition also depend on the physician's judgment and are unique to each individual. Factors influencing dosage include the physical and clinical condition of the subject, the route of administration, the goal of treatment (relief of symptoms during therapy), and the efficacy, stability, and toxicity of specific compositions. After preparation, the solutions will be administered in a manner compatible with the dosage and in an amount that is therapeutically or prophylactically effective. The compositions are easily administered in various dosage forms, such as the injection solutions described in this application.

A "biologically active agent" or active agent disclosed herein refers to one or more isolated or recombinant polypeptides, isolated or recombinant polynucleotides, vectors, isolated host cells, or antibodies, as well as compositions containing one or more of these agents.

"Administration" may be carried out as a single dose, continuously, or periodically throughout the course of treatment. Methods for determining the most effective agents and the dosage of administration are known to those skilled in the art and will vary depending on the composition used for therapy, the purpose of therapy, the target cell being treated, and the subject being treated. Single or multiple administrations may be performed, with the dose level and nature of the dose being selected by the attending physician. Suitable dosage forms and routes of administration of the agents are known in the art. The route of administration may also be determined. The method for determining the most effective route of administration is known to those skilled in the art, and such route will vary depending on the composition used for treatment, the purpose of treatment, the health condition or stage of the disease of the subject being treated, and the target cell or tissue. Non-limiting examples of routes of administration include oral administration, nasal administration, injection, and topical application.

The agent of the present invention can be administered for therapeutic purposes by any suitable route. It is also understood that the optimal route will vary depending on the condition and age of the recipient, as well as the disease being treated.

As used herein, the term "contacting" refers to the direct or indirect binding or interaction between two or more molecules. A specific example of a direct interaction is binding. A specific example of an indirect interaction is when one entity acts on a mediator molecule, which in turn acts on a second entity. The term "contacting," as used herein, includes contacting in solution, on a solid phase, in vitro, ex vivo, in a cell, and in vivo. Contacting in vivo may be referred to as administering or administering.

The term "effective amount" refers to an amount sufficient to achieve the desired effect. In the context of therapeutic or prophylactic applications, an effective amount will depend on the type and severity of the particular condition and the characteristics of the individual subject, such as general health, age, gender, body weight, and tolerance to the pharmaceutical compositions. With respect to an immunogenic composition, in some embodiments, an effective amount is an amount sufficient to provide a protective response against a pathogen. In other embodiments, an effective amount of an immunogenic composition is an amount sufficient to generate antibodies against the antigen. In some embodiments, an effective amount is the amount necessary to generate passive immunity in a subject in need thereof. For immunogenic compositions, in some embodiments, an effective amount will depend on the intended use, the degree of immunogenicity of the particular antigenic compound, and the health/responsiveness of the subject's immune system, in addition to the factors described above. A skilled artisan will be able to determine appropriate amounts based on these and other factors.

For in vitro use, in some embodiments, the effective amount will depend on the scale and nature of the intended application. It will also depend on the nature and sensitivity of the in vitro target and the methods used. A skilled artisan will be able to determine the effective amount based on these and other considerations. The effective amount may involve one or more administrations of the composition, depending on the embodiment of the invention.

The term "contact" refers to the direct or indirect association or interaction between two or more molecules. A specific example of a direct interaction is binding. A specific example of an indirect interaction is when one entity acts on a mediator molecule, which in turn acts on a second entity. The term "contact," as used herein, includes contact in solution, on a solid phase, in vitro, ex vivo, within a cell, and in vivo. In vivo contact may be referred to as administration or administration.

The term "conjugated moiety" refers to a moiety that can be added to an isolated chimeric polypeptide by forming a covalent bond with a residue of the chimeric polypeptide. The moiety can be linked directly to a residue of the chimeric polypeptide or can form a covalent bond with a linker, which in turn forms a covalent bond with a residue of the chimeric polypeptide.

A "peptide conjugate" refers to the association, via covalent or non-covalent linkage, of one or more polypeptides and another chemical or biological compound. In a non-limiting example, "conjugation" of a polypeptide to a chemical compound results in increased stability or efficacy of the polypeptide for its intended use. In one embodiment, the peptide is conjugated to a carrier, wherein the carrier is a liposome, a micelle, or a pharmaceutically acceptable polymer.

Liposomes are microscopic vesicles composed of concentric lipid bilayers. Structurally, liposomes vary in size and shape from long tubes to spherical structures with dimensions ranging from several hundred angstroms to fractions of a millimeter. The vesicle-forming lipids are selected to achieve a desired degree of fluidity or rigidity of the final complex, providing the lipid composition of the outer layer. They are neutral (cholesterol) or bipolar and include phospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), and sphingomyelin (SM), as well as other types of bipolar lipids, including, but not limited to, dioleoylphosphatidylethanolamine (DOPE), with hydrocarbon chain lengths in the range of 14–22 and saturated or with one or more C=C double bonds. Examples of lipids capable of forming stable liposomes, alone or in combination with other lipid components, are phospholipids such as hydrogenated soy phosphatidylcholine (HSPC), lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cephalin, cardiolipin, phosphatidic acid, cerebrosides, distearoylphosphatidylethanolamine (DSPE), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), palmitoylteoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), and dioleoylphosphatidylethanolamine 4-(N-maleimidotriethyl)cyclohexane-1-carboxylate (DOPE-mal). Additional phosphorus-free lipids that can be incorporated into liposomes include stearylamine, dodecylamine, hexadecylamine, isopropyl myristate, triethanolamine lauryl sulfate, alkyl aryl sulfate, acetyl palmitate, glycerol ricinoleate, hexadecyl stearate, amphoteric acrylic polymers, polyethyloxylated fatty acid amides, and the cationic lipids mentioned above (DDAB, DODAC, DMRIE, DMTAP, DOGS, DOTAP (DOTMA), DOSPA, DPTAP, DSTAP, DC-Chol). Negatively charged lipids include phosphatidic acid (PA), dipalmitoylphosphatidylglycerol (DPPG), ditheoylphosphatidylglycerol (DOPG), and dicetyl phosphate, which are capable of forming vesicles. Liposomes can generally be divided into three categories based on their overall size and the nature of their lamellar structure. Three classifications, developed at the New York Academy of Sciences meeting, "Liposomes and Their Applications in Biology and Medicine," December 1977, include multilamellar vesicles (MLVs), small unilamellar vesicles (SUVs), and large unilamellar vesicles (LUVs). Biologically active agents can be encapsulated in them for administration according to the methods described herein.

A micelle is an aggregate of surfactant molecules dispersed in a liquid colloid. A typical micelle in aqueous solution forms an aggregate with hydrophilic head regions in contact with the surrounding solvent, isolating hydrophobic tail regions in the center of the micelle. This type of micelle is known as a normal-phase micelle (oil-in-water micelle). Inverse micelles have head groups in the center with protruding tails (water-in-oil micelle). Micelles can be used to attach a polynucleotide, polypeptide, antibody, or composition described herein to facilitate their efficient delivery to a target cell or tissue.

The phrase "pharmaceutically acceptable polymer" refers to a group of compounds that can be conjugated to one or more polypeptides described herein. Conjugation of a polymer to a polypeptide is expected to increase the half-life of the polypeptide in vivo and in vitro. Non-limiting examples include polyethylene glycols, polyvinylpyrrolidones, polyvinyl alcohols, cellulose derivatives, polyacrylates, polymethacrylates, sugars, polyols, and mixtures thereof. Biologically active agents can be conjugated to a pharmaceutically acceptable polymer for administration according to the methods described herein.

A "gene delivery vehicle" is defined as any molecule that can transfer incorporated polynucleotides into a host cell. Examples of gene delivery vehicles include liposomes, micelles, biocompatible polymers, including natural polymers and synthetic polymers; lipoproteins; polypeptides; polysaccharides; lipopolysaccharides; artificial viral envelopes; metal particles; and bacteria or viruses such as baculovirus, adenovirus, and retrovirus, bacteriophage, cosmid, plasmid, fungal vectors, and other recombinant vehicles commonly used in the art that have been described for expression in a variety of eukaryotic and prokaryotic hosts and can be used for gene therapy as well as simple protein expression.

The polynucleotide disclosed in this application can be delivered to a cell or tissue or organism of a subject using a gene delivery vehicle. "Gene delivery," "gene transfer," "transduction," and the like, as used herein, are terms that refer to the introduction of an exogenous polynucleotide (sometimes referred to as a "transgene") into a host cell, regardless of the method used for introduction. Such methods include a variety of well-known techniques, such as gene transfer using vectors (by, for example, viral infection/transfection or various other protein- or lipid-based gene delivery complexes), as well as technologies that facilitate the delivery of "naked" polynucleotides (such as electroporation, "gene gun" delivery, and various other technologies used to introduce polynucleotides). The introduced polynucleotide can be stably or transiently maintained in the host cell. Stable maintenance typically requires that the introduced polynucleotide either contain an origin of replication compatible with the host cell or integrate into a host cell replicon, such as an extrachromosomal replicon (e.g., a plasmid), a nuclear chromosome, or a mitochondrial chromosome. A number of vectors are capable of mediating gene transfer into mammalian cells, as is known in the art and described herein.

As used herein, the term "cfDNA" refers to extracellular DNA found as a component of pathogenic biofilms.

A plasmid is an extrachromosomal DNA molecule, separate from chromosomal DNA, capable of replication independently of chromosomal DNA. In many cases, it is circular and double-stranded. Plasmids provide a mechanism for horizontal gene transfer within a microbial population and typically confer a selective advantage under given environmental conditions. Plasmids can carry genes that confer resistance to natural antibiotics in a competitive ecological niche, or alternatively, the proteins they produce can act as toxins under similar circumstances.

Plasmids used in genetic engineering are called plasmid vectors. Many plasmids are commercially available for such applications. The gene to be replicated is inserted into copies of the plasmid, which contains genes that render cells resistant to specific antibiotics and a multiple cloning site (MCS, or polylinker), which is a short region containing several commonly used restriction sites that allow DNA fragments to be easily inserted at this location. Another important use of plasmids is the production of large quantities of proteins. In this case, researchers grow bacteria containing a plasmid carrying the gene of interest. Just as bacteria produce proteins that confer antibiotic resistance, they can also be induced to produce large quantities of proteins based on the inserted gene. This is a cheap and easy way to mass-produce a gene or the protein encoded by that gene.

Yeast artificial chromosome, or YAC, refers to a vector used for cloning large DNA fragments (greater than 100 kb and up to 3,000 kb). It is an artificially constructed chromosome containing telomeric, centromeric, and origin of replication sequences necessary for replication and maintenance in yeast cells. Constructed using a circular plasmid, they are linearized using restriction enzymes, and DNA ligase can then add the desired sequence or gene to the linear molecule using sticky ends. Yeast expression vectors such as YAC, YIp (yeast integrating plasmid) and YEps (yeast episomal plasmid) are extremely useful because eukaryotic protein products with post-translational modifications can be produced, as yeast itself is a eukaryotic cell, however, YACs have been found to be more unstable than BACs, causing chimeric effects.

A "viral vector" is defined as a recombinantly produced virus or viral particle that contains a polynucleotide for delivery into a host cell either in vivo, ex vivo, or in vitro. Examples of viral vectors include retroviral vectors, adenoviral vectors, adeno-associated virus vectors, alphavirus vectors, etc. Infectious tobacco mosaic virus (TMV)-based vectors can be used to produce proteins and have been reported to express the griffithsin protein in tobacco leaves (O'Keefe et al. (2009) Proc. Nat. Acad. Sci. USA 106(15):6099–6104). Alphavirus vectors, such as Semliki Forest virus-based vectors and Sindbis virus-based vectors, have also been generated for use in gene therapy and immunotherapy. See Schlesinger & Dubensky (1999) Curr. Opin. Biotechnol. 5:434–439 and Ying et al. 16:95–118; Eren et al. Med. 5(7):823–827. In cases where gene transfer is mediated by a retroviral vector, the vector construct refers to a polynucleotide containing the retroviral genome, or a portion thereof, and the therapeutic gene.

As used herein, the terms "retroviral-mediated gene transfer" or "retroviral transduction" have the same meaning and refer to the process by which a gene or nucleic acid sequence is stably transferred into a host cell through viral entry and integration of its genome into the host cell's genome. The virus may enter the host cell via its normal infection mechanism or may be modified to bind to a different receptor or ligand on the host cell surface for entry. As used herein, the term "retroviral vector" refers to a viral particle capable of introducing exogenous nucleic acid into a cell via a viral or virus-like entry mechanism.

Retroviruses carry their genetic information in the form of RNA; however, once the virus infects a cell, the RNA is reverse-transcribed into DNA, which is integrated into the genomic DNA of the infected cell. The integrated form of DNA is called a provirus.

When gene transfer is mediated by a DNA viral vector such as an adenovirus (Ad) or adeno-associated virus (AAV), the vector construct refers to a polynucleotide containing the viral genome or a portion thereof and a transgene. Adenoviruses (Ad) are a relatively well-characterized, homogeneous group of viruses comprising more than 50 serotypes. See, for example, published international application WO 95/27071. Adenoviruses do not require integration into the host cell genome. Vectors based on recombinant Ads have also been constructed, including those that reduce the potential for recombination and the generation of wild-type virus; see published international applications WO 95/00655 and WO 95/11984. Wild-type AAV is highly infective and specific when integrated into the host cell genome, see Hermonat & Muzyczka (1984) Proc. Natl. Acad. Sci. USA 81:6466–6470 and Lebkowski et al. (1988) Mol. Cell. Biol. K8:3988–3996.

Vectors that contain both a promoter and a cloning site into which a polynucleotide can be operably inserted are well known in the art. Such vectors are capable of transcribing RNA in vitro or in vivo and are commercially available from companies such as Stratagene (La Jolla, CA) and Promega Biotech (Madison, WI). Optimizing in vitro expression and/or transcription may require removal, addition, or modification of the 5' and/or 3' untranslated portions of clones to eliminate additional, potentially inappropriate alternative translation initiation codons or other sequences that may interfere with expression or reduce expression at the transcriptional or translational level. Alternatively, consensus ribosome binding sites can be inserted immediately 5' of the start codon to enhance expression.

Gene delivery vehicles also include DNA/liposome complexes, micelles, and targeted viral protein-DNA complexes. Liposomes, which also contain a targeting antibody or fragment thereof, can be used in the methods disclosed herein. In addition to delivering polynucleotides to a cell or cell population, direct introduction of the proteins described herein into a cell or cell population can be accomplished using a non-limiting protein transfection method, alternatively under culture conditions that can enhance expression and/or promote the activity of the proteins disclosed herein.

As used herein, the terms "antibody," "antibodies," and "immunoglobulin" include whole antibodies and any antigen-binding fragment or single chain thereof. Thus, the term "antibody" includes any molecule containing a protein or peptide that contains at least a portion of an immunoglobulin molecule. The terms "antibody," "antibodies," and "immunoglobulin" also include immunoglobulins of any isotype, antibody fragments that retain specific binding to an antigen, including, but not limited to, Fab, Fab', F(ab)2, Fv, scFv, dsFv, Fd fragments, dAb, VH, VL, VhH, and V-NAR domains; minibodies, diabodies, tribodies, tetrabodies, and kappa bodies; multispecific antibody fragments constructed from antibody fragments. Examples of such include, but are not limited to, the antigen-binding complementarity determining region (CDR) of a heavy or light chain or a ligand-binding portion thereof, the variable region of a heavy or light chain (also referred to herein as a variable domain), the constant region of a heavy chain or light chain (also referred to herein as a constant domain), the framework (FR) region or any portion thereof, at least one portion of a binding protein, chimeric antibodies, humanized antibodies, single-chain antibodies, and fusion proteins comprising an antigen-binding portion of an antibody and a non-antibody protein. The variable regions of the heavy and light chains of an immunoglobulin molecule include a binding domain that interacts with an antigen. The constant regions of antibodies (Ab) can mediate the binding of immunoglobulin to host tissues. The term "anti-" used before a protein name, such as anti-DNABII, anti-IHF, anti-HU, or anti-OMP P5, refers to a monoclonal or polyclonal antibody that binds to and/or has affinity for a specific protein. For example, "anti-IHF" refers to an antibody that binds to the IHF protein. A specific antibody may have affinity for or bind to proteins other than the protein it was raised against. For example, although anti-IHF is specifically raised against the IHF protein, it may also bind other proteins that are related due to either sequence homology or structural homology.

Complementarity-determining regions (CDRs) are part of the variable region of an antibody or T-cell receptor, generated by B cells and T cells, respectively, and these molecules bind to their specific antigen (also called an epitope). In some embodiments, the terms "variable region" and "variable domain" are used interchangeably, referring to a light or heavy chain polypeptide of an antibody that varies significantly in amino acid sequence between different antibodies and determines the conformation of the binding site that confers specificity to a particular antigen. In another embodiment, the variable region has a length of from about 90 amino acids to about 200 amino acids, including, but not limited to, a length of about 100 amino acids, or alternatively a length of about 110 amino acids, or alternatively a length of about 120 amino acids, or alternatively about 130 amino acids, or alternatively about 140 amino acids, or alternatively about 150 amino acids, or alternatively about 160 amino acids, or alternatively about 170 amino acids, or alternatively about 180 amino acids, or alternatively about 190 amino acids. In some embodiments, the variable region of an amino acid sequence disclosed herein refers to about the first 100 amino acids, or alternatively about 110 amino acids, or alternatively about 120 amino acids, or alternatively about 130 amino acids, or alternatively about 140 amino acids, or alternatively about 150 amino acids of the sequence (including or excluding a signal peptide, if applicable).

A set of CDRs constitutes a paratope, also called an antigen-binding site, which is the part of an antibody that recognizes and binds to an antigen. There are three CDR regions (CDR1, CDR2, and CDR3) arranged non-sequentially, optionally from the amino terminus to the carboxy terminus, in the amino acid sequence of a variable region of an antigen receptor, such as a heavy chain or a light chain. As used herein, the designation CDRn refers to CDRn in an immunoglobulin chain or regions derived from an immunoglobulin chain, wherein the number n is selected from 1-3. In one embodiment, CDRLn refers to CDRn in an immunoglobulin light chain or regions derived from a light chain, wherein the number n is selected from 1-3; while CDRHn refers to CDRn in an immunoglobulin heavy chain or regions derived from a heavy chain, wherein the number n is selected from 1-3. In some embodiments, the framework region (FR) refers to the portion of the variable region that is not a CDR. In some embodiments, FRn refers to the FR of a heavy chain or a light chain, or a region derived from a heavy chain or a light chain, wherein the number n is selected from 1-4. In some embodiments, the variable region comprises or consists essentially of the following elements (optionally in the order listed, and optionally also from amino-terminus to carboxy-terminus): FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

The variable regions and/or CDRs of an antibody or fragment thereof can be determined by one skilled in the art, for example, using publicly available or commercially available tools. Non-limiting examples of such tools include IgBlast (available at www.ncbi.nlm.nih.gov/igblast/), Scaligner (available from Drugdesigntech at www.scaligner.com/), IMGT rules and/or tools (see, for example, www.imgt.org/IMGTScientificChart/Nomenclature/IMGT-FRCDRdefinition.html, also available at www.imgt.org/), Chothia Canonical Assignment (available at www.bioinf.org.uk/abs/chothia.html), Antigen Receptor Numbering and Receptor Classification (ANARCI, available at opig.stats.ox.ac.uk/webapps/newsabdab/sabpred/anarci/), Kabat numbering method/scheme (e.g., Kabat, E.A., et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242) or the Paratome web server (available at www.ofranlab.org/paratome/, see Vered Kunik, et al, Nucleic Acids Research, Volume 40, Issue W1, 1 July 2012, Pages W521–W524).

Antibodies can be polyclonal, monoclonal, multispecific (e.g., bispecific antibodies), or antibody fragments, provided they exhibit the desired biological activity. Antibodies can be isolated from any suitable biological source, such as mice, rats, sheep, and dogs.

The terms "polyclonal antibody" or "polyclonal antibody composition" as used herein refer to a preparation of antibodies derived from various B-cell lines. They represent a mixture of immunoglobulin molecules secreted against a specific antigen, each recognizing a distinct epitope.

As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies. Monoclonal antibodies are highly specific because each monoclonal antibody is directed against a single antigenic determinant. Antibodies can be labeled with a detectable label, such as a radioisotope, an enzyme that produces a detectable product, a fluorescent protein, etc. Antibodies can be further conjugated to other moieties, such as members of specific binding pairs, such as biotin (a member of the biotin-avidin specific binding pair), etc. Antibodies can also be bound to a solid support, including, but not limited to, polystyrene sheets or beads, etc.

Monoclonal antibodies can be produced using hybridoma technology or recombinant DNA techniques known in the art. A hybridoma is a cell produced in the laboratory by fusing an antibody-producing lymphocyte with a non-antibody-producing malignant tumor cell, typically myeloma or lymphoma. The hybridoma replicates and produces a constant pattern of a specific monoclonal antibody. Alternative methods for producing or selecting antibodies include exposing lymphocytes to antigens of interest in vitro and screening antibody display libraries in cells, phages, or similar systems.

The term "human antibody" as used herein is intended to characterize antibodies comprising variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies disclosed herein may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or somatic mutation in vivo). However, the term "human antibody" as used herein does not include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. Thus, as used herein, the term "human antibody" refers to an antibody in which substantially every portion of the protein (e.g., CDRs, framework region, CL, CH domains (e.g., CH1, CH2, CH3), hinge region (VL, VH)) is substantially non-immunogenic in humans and has only minor sequence changes or variations. Similarly, antibodies referred to as primate (marmoset, baboons, chimpanzees, etc.), rodent (mice, rats, rabbits, guinea pigs, hamsters, etc.) and other mammalian antibodies refer to specific antibodies of certain species, subgenera, genera, subfamilies, families. Furthermore, chimeric antibodies include any combination of the above. Such changes or variations do not necessarily maintain or reduce immunogenicity in humans or other species compared to unmodified antibodies. Thus, a human antibody differs from a chimeric or humanized antibody. It is known that a human antibody can be produced in an animal organism or by a prokaryotic or eukaryotic cell capable of expressing functionally rearranged human immunoglobulin genes (e.g., heavy chain and/or light chain). Furthermore, when a human antibody is a single-chain antibody, it may contain a linker peptide that is not found in native human antibodies. For example, Fv may contain a linker peptide, e.g., two to about eight glycine residues or other amino acids, that connects the variable region of the heavy chain and the variable region of the light chain. Such linker peptides are considered to be of human origin.

As used herein, the term "a human antibody is derived from" a particular germline sequence if the antibody is obtained from a system that utilizes human immunoglobulin sequences, such as by immunizing a transgenic mouse carrying human immunoglobulin genes or by screening a human immunoglobulin gene library. A human antibody that is "derived" from a human germline immunoglobulin sequence can be identified as such by comparing the amino acid sequence of the human antibody to the amino acid sequence of human germline immunoglobulins. The amino acid sequence of a selected human antibody is typically at least 90% identical to the amino acid sequence encoded by the human germline immunoglobulin gene and contains amino acid residues that identify the antibody as human when compared to the amino acid sequences of germline immunoglobulins from other species (e.g., mouse germline immunoglobulin sequences). In some cases, the amino acid sequence of a human antibody may be at least 95% identical, or even at least 96%, 97%, 98%, or 99% identical, to the amino acid sequence encoded by the germline immunoglobulin gene. Typically, a human antibody derived from a particular human germline sequence will have no more than 10 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene. In some cases, a human antibody may differ by no more than 5 amino acids, or even no more than 4, 3, 2, or 1 amino acid from the amino acid sequence encoded by the germline immunoglobulin gene.

As used herein, the term "humanized antibody" or "humanized immunoglobulin" refers to a human/non-human chimeric antibody that contains a minimal sequence derived from a non-human immunoglobulin. In general, humanized antibodies are human immunoglobulins (the recipient antibody) in which residues of the variable region or a fragment thereof (e.g., 1, 2, 3, 4, 5, or all 6 CDRs) of the recipient are replaced with residues from the variable region or a fragment thereof (e.g., 1, 2, 3, 4, 5, or all 6 CDRs) of a non-human antibody (the donor antibody), such as, for example, a mouse, rat, rabbit, or non-human primate antibody, having the desired specificity, affinity, and properties. Humanized antibodies may contain amino acid residues that are not found in the recipient antibody or in the donor antibody. A humanized antibody may also optionally contain at least a portion of an immunoglobulin constant region (Fc), typically a human immunoglobulin or a non-human antibody, comprising one or more amino acids in the framework, constant region, or CDR that have been replaced with appropriately positioned amino acids from a human antibody. Without wishing to be bound by any theory, the inventors note that humanized antibodies elicit a reduced immune response in a human host compared to a non-humanized version of the same antibody. Humanized antibodies may have conservative amino acid substitutions that do not substantially affect antigen binding or other antibody functions. Conservative substitution groups include: glycine-alanine, valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, serine-threonine, and asparagine-glutamine. In particular, the humanized antibodies disclosed herein specifically bind to a DNABII polypeptide or a fragment thereof (such as a head chimeric peptide or a tail chimeric peptide) within a certain range of one or more of the following values: _EC50 , _Kon , _Koff , _KA , and/or _KD , and inhibit or release certain cytokines when treating a subject. In another embodiment, a humanized antibody that specifically binds to the head region of a DNABII polypeptide (such as a head chimeric peptide) disrupts biofilm both in vivo and in vitro. Furthermore, the humanization process, although a rational way to engineer antibodies, may result in unexpected changes (positive or negative) in, for example, binding affinity, antigen specificity, or physical properties such as solubility or aggregation ability; therefore, the properties of humanized antibodies cannot be predicted based on the properties of the parent non-human antibody.

In one embodiment, the antibody used in the present invention may be a recombinant antibody. The term "recombinant antibody" as used herein includes all antibodies that are produced, expressed, created, or isolated by recombinant methods, such as antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for immunoglobulin genes, or produced by a hybridoma obtained using an animal, antibodies isolated from a host cell transformed to express an antibody, for example, from a transfectoma, antibodies isolated from a library of recombinant, combinatorial antibodies, and antibodies produced, expressed, created, or isolated by any other methods that involve splicing an immunoglobulin (Ig) gene sequence with other DNA sequences. However, in some embodiments, such recombinant antibodies may be subjected to in vitro mutagenesis (or, when an animal transgenic for Ig sequences is used, somatic mutagenesis in vivo), and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies represent sequences that may not naturally exist in the germline repertoire of antibodies in vivo. Methods for producing these antibodies are described herein.

In one embodiment, the antibody used in the present invention may be a chimeric antibody. As used herein, the term "chimeric antibodies" refers to antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, based on the variable and constant region genes of antibodies from different species.

As used herein, the term "antibody derivative" encompasses a full-length antibody or antibody fragment wherein one or more amino acids are chemically modified by alkylation, PEGylation, acylation, ester formation, amide formation, or the like, for example, to link the antibody to a second molecule. Such technology includes, but is not limited to, PEGylated antibodies, cysteine-PEGylated antibodies, and variants thereof.

As used herein, the term "label" means a directly or indirectly detectable compound or composition that is directly or indirectly conjugated to a detectable entity, such as N-terminal histidine tags (N-His), magnetically active isotopes such as ¹¹⁵ Sn, ¹¹⁷ Sn, and ¹¹⁹ Sn, non-radioactive isotopes such as ¹³ C and ¹⁵ N, a polynucleotide, or a protein such as an antibody, to create a "labeled" entity. The term also includes sequences conjugated to the polynucleotide that will provide a signal upon expression of integrated sequences such as green fluorescent protein (GFP), etc. The label may be detectable in itself (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze a detectable chemical change in a substrate compound or composition. Labels may be suitable for small-scale detection or more suitable for high-throughput screening. Suitable labels thus include, but are not limited to, magnetically active isotopes, non-radioactive isotopes, radioisotopes, fluorochromes, chemiluminescent compounds, dyes, and proteins, including enzymes. A label may be simply detectable or quantifiable. A response that is simply detected typically involves a response whose existence is merely confirmed, whereas a quantifiable response typically has a quantifiable value (e.g., a numerical value), such as intensity, polarization, and/or other properties. In luminescence or fluorescence assays, the detectable response may be obtained directly using a phosphor or fluorophore linked to the assay component actually involved in binding, or indirectly using a phosphor or fluorophore linked to another (e.g., a reporter or indicator) component. Examples of luminescent labels that generate signals include, but are not limited to, bioluminescent and chemiluminescent labels. The detectable luminescence response typically involves a change or occurrence of a luminescence signal. Suitable methods and luminophores for luminescently labeling assay components are known in the art and are described, for example, in Haugland, Richard P. (1996) Handbook of Fluorescent Probes and Research Chemicals (6th ed). Examples of luminescent probes include, but are not limited to, aequorin and luciferases.

As used herein, the term "immunoconjugate" encompasses an antibody or antibody derivative associated or linked to a second agent, such as a cytotoxic agent, a detectable agent, a radioactive agent, a targeting agent, a human antibody, a humanized antibody, a chimeric antibody, a synthetic antibody, a semi-synthetic antibody, or a multispecific antibody.

Examples of suitable fluorescent labels include, but are not limited to, fluorescein, rhodamine, tetramethylrhodamine, eosin, erythrosine, coumarin, methylcoumarins, pyrene, malachite green, stilbene, Lucifer Yellow, Cascade Blue™, and Texas Red. Other suitable optical dyes are described in Haugland, Richard P. (1996) Handbook of Fluorescent Probes and Research Chemicals (6th ed.).

In another embodiment, the fluorescent label is functionalized to facilitate covalent attachment to a cellular component present in or on the surface of a cell or tissue, such as a cell surface marker. Suitable functional groups include, but are not limited to, isothiocyanate groups, amino groups, haloacetyl groups, maleimides, succinimidyl esters, and sulfonyl halides, all of which can be used to attach the fluorescent label to a second molecule. The choice of functional group on the fluorescent label will depend on the site of attachment to the linker, agent, marker, or second labeling agent.

The term "eukaryotic cells" refers to all biological kingdoms except Monera. They are easily distinguished by their membrane-enclosed nucleus. Animals, plants, fungi, and protozoa are eukaryotes, or organisms whose cells are organized into complex structures by internal membranes and a cytoskeleton. The most characteristic membrane-containing structure is the nucleus. Unless otherwise specified, the term "host" encompasses the eukaryotic host, including, for example, yeast, higher plant cells, insects, and mammals. Non-limiting examples of eukaryotic cells or host cells include those of monkeys, cows, pigs, mice, rats, birds, reptiles, and humans.

Prokaryotic cells, which typically lack a nucleus or other membrane-bound organelles, are divided into two domains: bacteria and archaea. In addition to chromosomal DNA, these cells may also contain genetic information in a circular molecule called an episome. Bacterial cells are very small, approximately the size of an animal mitochondria (about 1-2 µm in diameter and 10 µm in length). Prokaryotic cells have three basic shapes: rod-shaped, spherical, and helical. Rather than undergoing complex replication processes like eukaryotes, bacterial cells reproduce through binary fission. Examples include, but are not limited to, Bacillus, E. coli, and Salmonella.

A "native" or "natural" antigen is a polypeptide, protein, or fragment containing an epitope that has been isolated from a natural biological source and that can specifically bind to an antigen receptor, particularly the T-cell antigen receptor (TCR), of a subject.

The terms "antigen" and "antigenic" refer to molecules capable of being recognized by an antibody or otherwise acting as a member of an antibody-ligand pair. "Specific binding" or "binding" refers to the interaction of the antigen with the variable regions of the heavy and light chains of immunoglobulin. Antibody-antigen binding can occur in vivo or in vitro. One skilled in the art understands that macromolecules, including proteins, nucleic acids, fatty acids, lipids, lipopolysaccharides, and polysaccharides, have the potential to act as an antigen. One skilled in the art also understands that nucleic acids encoding a protein potentially capable of acting as an antibody ligand necessarily encode an antigen. One also understands that antigens are not limited to full-length molecules but may also include portions of molecules. The term "antigenic" refers to molecules that exhibit the properties of an antigen. It encompasses substances that are immunogenic, i.e., immunogens, as well as substances that cause immunological insensitivity or anergy, i.e. anergens.

An "altered antigen" is an antigen whose primary sequence differs from that of the corresponding wild-type antigen. Altered antigens may be produced by synthetic or recombinant methods and include, but are not limited to, antigenic peptides that are differentially modified during or after translation, such as by phosphorylation, glycosylation, cross-linking, acylation, proteolytic cleavage, association with an antibody molecule, a membrane molecule, or another ligand. (Ferguson et al. (1988) Ann. Rev. Biochem. 57:285-320). The synthetic or altered antigen described herein is designed to bind to the same TCR as the natural epitope.

A "self-antigen," also referred to herein as a native antigen or wild-type antigen, is an antigenic peptide that induces little or no immune response in a subject due to self-tolerance to the antigen. An example of a self-antigen is the melanoma-specific antigen gp100.

"Immune response" broadly refers to antigen-specific responses of lymphocytes to foreign substances. The terms "immunogen" and "immunogenic" refer to molecules capable of eliciting an immune response. All immunogens are antigens, but not all antigens are immunogenic. The immune response described herein may be humoral (mediated by antibody activity) or cell-mediated (mediated by T-cell activation). The response may occur in vivo or in vitro. One skilled in the art understands that various macromolecules, including proteins, nucleic acids, fatty acids, lipids, lipopolysaccharides, and polysaccharides, have the potential to be immunogenic. One skilled in the art also understands that nucleic acids encoding a molecule capable of eliciting an immune response necessarily encode an immunogen. One skilled in the art also understands that immunogens are not limited to full-length molecules but may include portions of molecules.

The term "passive immunity" refers to the transfer of immunity from one subject to another through the transfer of antibodies. Passive immunity can arise naturally, for example, when maternal antibodies are transferred to a fetus. Passive immunity can also arise artificially, for example, when antibody compositions are administered to non-immune subjects. Antibody donors and recipients can be humans or non-human subjects. Antibodies can be polyclonal or monoclonal, can be produced in vitro or in vivo, and can be purified, partially purified, or unpurified, depending on the embodiment of the invention. In some embodiments described herein, passive immunity is provided to a subject in need thereof by administering antibodies or antigen-binding fragments that specifically recognize or bind to a particular antigen. In some embodiments, passive immunity is provided by administering an isolated or recombinant polynucleotide encoding an antibody or antigen-binding fragment that specifically recognizes or binds to a particular antigen.

As used herein, a "ligand" is a polypeptide. In one embodiment, the term "ligand" as used herein refers to any molecule that binds to a specific site on another molecule. In other words, a ligand is a specific protein that reacts with an immune effector cell, an antibody, or DNA. In one aspect, the ligand site on the protein directly binds to a complementary binding site on the immune effector cell.

As used herein, the terms "solid-phase carrier" or "solid support" are interchangeable and are not limited to a specific type of carrier. A wide variety of supports are available, known to those skilled in the art. Solid-phase supports include silica gels, resins, derivatized plastic films, glass beads, cotton, plastic beads, and aluminum oxide gels. The term "solid support" as used herein also includes synthetic antigen-presenting matrices, cells, and liposomes. A suitable solid-phase support can be selected based on the desired end use and suitability for various protocols. For example, for peptide synthesis, the solid phase support may be a resin such as polystyrene (e.g., PAM resin, obtained from Bachem Inc., Peninsula Laboratories, etc.), POLYHIPE® resin (obtained from Aminotech, Canada), polyamide resin (obtained from Peninsula Laboratories), polyethylene glycol-grafted polystyrene resin (TentaGel®, Rapp Polymere, Tuebingen, Germany), or polydimethylacrylamide resin (obtained from Milligen/Biosearch, California).

Examples of solid-phase supports include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylases, natural and modified celluloses, polyacrylamides, gabbro, and magnetite. The support may be either somewhat soluble or insoluble. The support material can have virtually any possible structural configuration, provided that the bound molecule is capable of binding to the polynucleotide, polypeptide, or antibody. Thus, the support configuration may be spherical, as in a bead, or cylindrical, as in the inner surface of a test tube or the outer surface of a rod. Alternatively, the surface may be flat, such as a sheet, test strip, etc., or, alternatively, polystyrene beads. Those skilled in the art will recognize many other suitable supports for antibody or antigen binding or will be able to determine them through routine experimentation.

The biological sample or specimen used in the present invention may be obtained from a subject or may be a cell line sample or cultured cells or tissues. Examples of samples include, but are not limited to, a cell sample, a tissue sample, liquid samples such as blood and other liquid samples of biological origin (including, but not limited to, ocular fluids (aqueous humor and vitreous humor), peripheral blood, serum, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, intraocular fluid, amniotic fluid, earwax, breast milk, bronchoalveolar lavage fluid, semen, prostate fluid, Cooper's fluid or pre-ejaculate, female ejaculate, sweat, tears, cystic fluid, pleural and peritoneal fluids, pericardial fluid, ascites, lymph, chyme, chyle, bile, interstitial fluid, menstrual blood, pus, sebum, vomit, vaginal secretions/washings, synovial fluid, mucous secretions, fecal fluid, pancreatic juice, sinus lavage fluid, bronchopulmonary aspirates, blastula cyst fluid, or umbilical cord blood. In one embodiment, the biological sample is suspected of containing a biofilm. In another embodiment, the biological sample contains a biofilm.

As used herein, the term "signal peptide" or "signal polypeptide" encompasses an amino acid sequence typically found at the N-terminus of newly synthesized secretory or membrane polypeptides or proteins. The signal peptide functions to direct the polypeptide to a specific location in the cell, such as through the cell membrane, into the cell membrane, or into the nucleus. In some embodiments, the signal peptide is removed after localization. Examples of signal peptides are well known in the art. Non-limiting examples are described in U.S. Patent Nos. 8,853,381, 5,958,736, and 8,795,965.

As used herein, the term "chimera" or "chimeric peptide" refers to a recombinant polypeptide comprising or alternatively consisting essentially of or additionally consisting of two or more fragments or domains of a DNABII polypeptide conjugated directly or indirectly (e.g., via a linker) to each other. In one embodiment, the domains are conformational head domains and/or conformational tail domains. Additionally or alternatively, the two or more fragments or domains are derived from the same or different DNABII polypeptides. In one embodiment, the chimeric peptide comprises or alternatively consists essentially of or additionally consists of an IhfA head domain and an IhfB head domain conjugated directly or indirectly (e.g., via a linker) to each other. In another embodiment, the chimeric peptide comprises, or alternatively consists essentially or additionally consists of, an IhfA tail domain and an IhfB tail domain, conjugated directly or indirectly (e.g., via a linker) to each other. A "conformational head domain" of a polypeptide refers to a polypeptide that comprises a primary amino acid sequence, wherein the structure has an antiparallel beta-ribbon with a sharp turn, which is typically mediated by a proline residue. The "head" of the IHF polypeptide is shown in Figure 1 of WO 2018/129078.

In some embodiments, the IhfA5-mIhfB4NTHI chimeric head peptide comprises or consists essentially of or additionally consists of a polypeptide sequence

(SEQ ID NO: 38), wherein "X" is an optional amino acid linker sequence, optionally comprising or consisting essentially of or additionally consisting of from 1 to 20 amino acids; and wherein "X ₁ " is any amino acid or alternatively "X ₁ " is selected from the amino acids Q, R, K, S or T. In a further embodiment, "X ₁ " is K or Q. In yet another further embodiment, the head chimeric peptide IhfA5-mIhfB4NTHI comprises or consists essentially of or additionally consists of the polypeptide sequence

(SEQ ID NO: 39), wherein "X" is optionally a linker sequence of amino acids, optionally comprising or consisting essentially of or additionally consisting of from 1 to 20 amino acids. In another embodiment, the head chimeric peptide IhfA5-mIhfB4NTHI comprises or consists essentially of or additionally consists of a polypeptide sequence

(SEQ ID NO: 40).

In some embodiments, the IhfA3-IhfB2NTHI chimeric tail peptide comprises, consists essentially of, or additionally consists of the polypeptide sequence FLEEIRLSLESGQDVKLSGF-X-TLSAKEEIENMVKDILEFISQ (SEQ ID NO: 41), wherein "X" is an optional amino acid linker sequence, optionally comprising, or consisting essentially of, or additionally consisting of from 1 to 20 amino acids. In some embodiments, the linker is selected from any one or more of SEQ ID NOs: 42-49. In one embodiment, the IhfA3-IhfB2NTHI chimeric tail peptide comprises, consists essentially of, or additionally consists of FLEEIRLSLESGQDVKLSGFGPSLTLSAKEIENMVKDILEFISQ (SEQ ID NO: 50).

As used herein, the term "EC ₅₀ " refers to the concentration of an antibody or fragment thereof that halfway induces a response (e.g., binding between the antibody or fragment thereof and its target) between the baseline level and the maximum after a specified exposure time.

Several parameters are used in the present application to describe the reaction of binding and release of a receptor (R, such as an antibody or fragment thereof) and a ligand (L, such as the target of the antibody or fragment thereof) molecules, which is formally represented as: R + L ⊂ RL. The reaction is characterized by the association rate constant k _on and the dissociation rate constant k _off , which have the dimensions M ^-1 s ^-1 and s ^-1 , respectively. At equilibrium, the forward binding transition R + L → RL must be balanced by the reverse unbinding transition RL → R + L. That is, k _on [R] [L] = k _off [RL], where [R], [L], and [RL] represent the concentration of unbound free receptors, the concentration of unbound free ligand, and the concentration of receptor-ligand complexes. In addition, the equilibrium dissociation constant “K _D ” can be calculated as k _off /k _on , which is equal to [R] × [L]/[RL], and the equilibrium association constant “K _A ” can be calculated as k _on /k _off , which is equal to [RL]/([R] × [L]).

As used herein, the term "cytokine" refers to small proteins (approximately 5–20 kDa) important for cellular signaling, including, but not limited to, chemokines, interferons, interleukins (IL), lymphokines, and tumor necrosis factors, but not hormones. Cytokines are peptides and cannot penetrate the lipid bilayer of cells into the cytoplasm. An inflammatory cytokine, or proinflammatory cytokine, is a type of signaling molecule (cytokine) secreted by immune cells such as helper T cells (Th) and macrophages, as well as certain other cell types that promote inflammation. Cytokines include, but are not limited to, interleukin-1 (IL-1), IL-12, and IL-18, tumor necrosis factor alpha (TNF-α), gamma interferon (IFNγ), and granulocyte-macrophage colony-stimulating factor (GM-CSF) and play an important role in mediating the innate immune response. Inflammatory cytokines are predominantly produced and involved in amplifying inflammatory responses. The term "anti-inflammatory cytokines" includes immunoregulatory molecules that control the response of proinflammatory cytokines. Cytokines act in concert with specific cytokine inhibitors and soluble cytokine receptors to regulate the human immune response. The major anti-inflammatory cytokines include interleukin (IL)-1 receptor antagonist, IL-4, IL-6, IL-10, IL-11, and IL-13. Specific cytokine receptors for IL-1, tumor necrosis factor-alpha, and IL-18 also act as inhibitors of proinflammatory cytokines. Methods for measuring cytokine levels, including anti-inflammatory and proinflammatory cytokines, are well known in the art. For example, serum cytokine levels can be measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits.

As used herein, the term "anti-infective" refers to a drug that is capable of inhibiting the spread of an infectious organism or directly killing such an organism. This term includes, but is not limited to, antibiotics, fungicides, anthelmintics, antimalarials, antiprotozoals, antituberculosis agents, and antivirals. Fungicidal agents are also called antifungal agents. They kill or inactivate fungi and are used to treat fungal infections (including yeast infections). In one non-limiting example, polyene fungicidal agents are not absorbed when administered orally and are therefore used to treat fungal infections of the gastrointestinal tract, such as oral thrush. Other non-limiting examples include azole fungicidals, which are synthetic fungistatic agents with a broad spectrum of activity, echinocandins, which are lipopeptide molecules that non-competitively inhibit the enzyme (1,3) beta-d-glucan synthase and act on the fungal cell wall, fulvicin U/F (i.e., griseofulvin), grifulvin V (Pro) (i.e., griseofulvin), lamisil (Pro) (i.e., terbinafine), Gris-PEG (Pro) (i.e., griseofulvin), ancobon (Pro) (i.e., flucytosine), fulvicin P/G (i.e., griseofulvin), and terbinex (Pro) (i.e., terbinafine). Other anti-infective agents can be found, for example, at www.drugbank.ca/categories/DBCAT000065. The term "antiviral" or "antiviral" refers to a class of drugs used to treat viral infections. Most antiviral agents target specific viruses, while broad-spectrum antiviral agents are effective against a wide range of viruses. Unlike most antibiotics, antiviral agents do not destroy the target pathogen; instead, they inhibit its development. Some of the mechanisms by which they may act include preventing viral replication by inhibiting viral DNA polymerase; binding to specific cell surface receptors and inhibiting viral entry or membrane degradation; inhibiting viral protein synthesis; or blocking late stages of viral assembly. Non-limiting examples of antiviral agents can be found, for example, at www.drugbank.ca/categories/DBCAT000066.

As used herein, the term "antiparasitic agent" refers to a class of drugs indicated for the treatment of parasitic diseases, such as those caused by helminths, amoebae, ectoparasites, parasitic fungi, and protozoa, among others. Non-limiting examples of antiparasitic agents can be found, for example, at www.drugbank.ca/categories/DBCAT000522.

Methods for carrying out the invention

Antibody compositions

The present invention relates to an isolated antibody comprising a heavy chain variable domain (HC) sequence and a light chain variable domain (LC) sequence, wherein the immunoglobulin heavy chain and light chain variable domain sequences form an antigen-binding site that binds to an epitope of the DNABII protein. In some embodiments, the antibody or fragment thereof binds to a DNABII peptide (such as a DNABII peptide head region, including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI chimeric head peptide, and/or a DNABII peptide tail region, including, but not limited to: an IHF or HU tail region, an IHFA or IHFB tail region, and/or an IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the antibody or fragment thereof binds to the head chimeric peptide IhfA5-mIhfB4NTHI. In another embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2NTHI.

In one embodiment of the present invention there are provided antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 21-132 of SEQ ID NO: 7-9, 14 or 25, or (aa) 21-126 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of said.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a heavy chain (HC) comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 25-473 of SEQ ID NO: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and/or from a light chain sequence (LC) comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 21-239 of SEQ ID NO: 7-9, 14 or 25, or (aa) 21-233 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of said.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a heavy chain (HC) comprising, essentially consisting of, or consisting of a sequence selected from the group of sequences SEQ ID NO: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and/or a light chain (LC) sequence comprising, essentially consisting of, or consisting of a sequence selected from the group of SEQ ID NO: 7-12, 14, 25 or 27, or sequences equivalent to each of said.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 21-132 of SEQ ID NO: 6-9, 14 or 25, or (aa) 21-126 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of said.

In another embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of any one, any two, or all three CDR sequences selected from the group comprising the sequences of SEQ ID NOs: 1-6, 13, 24, or 26, or sequences equivalent to each of these; and/or any one, any two, or all three CDR sequences selected from the group comprising the sequences of SEQ ID NOs: 7-12, 14, 25, or 27, or sequences equivalent to each of these.

In one embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 25-144 of SEQ ID NO: 13, 24 or 26, or sequences equivalent to each of said; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 21-132 of SEQ ID NO: 14 or 25, or (aa) 21-126 of SEQ ID NO: 27, or sequences equivalent to each of said. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from the group of amino acid sequences (aa) 21-132 of SEQ ID NO: 7-9, 14 or 25, or (aa) 21-126 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of said.

Also provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 1, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from any one sequence of amino acids (aa) 21-132 of SEQ ID NO: 7-9, 14 or 25, or (aa) 21-126 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of the aforementioned.

In one embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 2, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence selected from any one of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 7-9, 14, or 25, or (aa) 21-126 of SEQ ID NO: 10-12, or 27, or sequences equivalent to each of the aforementioned. Also provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 3, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from any one sequence of amino acids (aa) 21-132 of SEQ ID NO: 7-9, 14 or 25, or (aa) 21-126 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of the aforementioned.

Also provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 4, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence selected from any one sequence of amino acids (aa) 21-132 of SEQ ID NO: 7-9, 14 or 25, or (aa) 21-126 of SEQ ID NO: 10-12 or 27, or sequences equivalent to each of the aforementioned.

In one embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 5, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence selected from any one of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 7-9, 14, or 25, or (aa) 21-126 of SEQ ID NO: 10-12, or 27, or sequences equivalent to each of the aforementioned. Also provided are antibodies and antigen-binding fragments thereof that comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 6, or a sequence equivalent thereto; and of a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence selected from any one of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 7-9, 14, or 25, or (aa) 21-126 of SEQ ID NO: 10-12, or 27, or sequences equivalent to each of the said

In one embodiment, antibodies and antigen-binding fragments thereof are provided that comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 25-144 of any of the sequences of SEQ ID NOs: 1-6, 13, 24, or 26, or sequences equivalent to each of said; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 21-132 of SEQ ID NO: 7, or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 25-144 of any of the sequences of SEQ ID NOs: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 21-132 of SEQ ID NO: 8, or a sequence equivalent to said. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 25-144 of any of the sequences of SEQ ID NOs: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 21-132 of SEQ ID NO: 9 or a sequence equivalent thereto.

In one embodiment, antibodies and antigen-binding fragments thereof are provided that comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 25-144 of any of the sequences of SEQ ID NOs: 1-6, 13, 24, or 26, or sequences equivalent to each of said sequences; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 21-126 of SEQ ID NO: 10, or a sequence equivalent to said sequence. Also provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 25-144 of any of the sequences of SEQ ID NOs: 1-6, 13, 24, or 26, or sequences equivalent to each of said; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 21-126 of SEQ ID NO: 11, or a sequence equivalent to said. Also provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 25-144 of any of the sequences of SEQ ID NO: 1-6, 13, 24 or 26, or sequences equivalent to each of said; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of a sequence of amino acids (aa) 21-126 of SEQ ID NO: 12 or a sequence equivalent to said.

Also provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO: 1, or a sequence equivalent thereto; and an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO: 7, or a sequence equivalent thereto. In one embodiment, provided are antibodies and antigen-binding fragments thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO: 1, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 8 or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 1, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 9 or a sequence equivalent thereto.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:2, or a sequence equivalent thereto; and from an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO:7, or a sequence equivalent thereto. In yet another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:2, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 8 or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 2, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO: 9 or a sequence equivalent thereto.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:3, or a sequence equivalent thereto; and from an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO:7, or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:3, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:8 or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO:3, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:9 or a sequence equivalent thereto.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:4, or a sequence equivalent thereto; and from an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-126 of SEQ ID NO:10, or a sequence equivalent thereto. In yet another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:4, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 11 or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 4, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 12 or a sequence equivalent thereto.

In one embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:5, or a sequence equivalent thereto; and from an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-126 of SEQ ID NO:10, or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:5, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 11 or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 5, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 12 or a sequence equivalent thereto.

In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:6, or a sequence equivalent thereto; and from an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-126 of SEQ ID NO:10, or a sequence equivalent thereto. In yet another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:6, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 11 or a sequence equivalent thereto. In another embodiment, antibodies and antigen-binding fragments thereof are provided, which comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 6, or a sequence equivalent thereto; and from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 12 or a sequence equivalent thereto.

In one embodiment, antibodies and fragments thereof are provided that comprise or essentially consist of or additionally consist of a sequence of a variable domain of an immunoglobulin heavy chain (HC) comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO:24, or a sequence equivalent thereto; and/or of a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:25, or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof binds to the head region of the DNABII peptide (including, but not limited to: the head region of IHF or HU, the head region of IHFA or IHFB, and/or the head chimeric peptide IhfA5-mIhfB4NTHI). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI head chimeric peptide. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, antibodies and fragments thereof are provided that comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO: 1, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO: 7, or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:1, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO:8, or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:1, or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:9 or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided that comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO:2 or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:7 or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:2, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO:8, or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:2, or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:9 or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided that comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO:3 or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:7 or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:3, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-132 of SEQ ID NO:8, or a sequence equivalent thereto. In another embodiment, antibodies and fragments thereof are provided, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:3, or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, consisting essentially of, or consisting of the sequence of amino acids (aa) 21-132 of SEQ ID NO:9 or a sequence equivalent thereto.

In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin, comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO:26, or a sequence equivalent thereto; and of a sequence of a variable domain of a light chain (LC) of an immunoglobulin, comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO:27, or a sequence equivalent thereto. In another embodiment, the antibody or a fragment thereof binds to the tail region of the DNABII peptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In one embodiment, the antibody or fragment thereof binds to the IhfA3-IhfB2NTHI tail chimeric peptide. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody and fragment thereof that comprise or essentially consist of or additionally consist of a sequence of a variable domain of an immunoglobulin heavy chain (HC) comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 4, or a sequence equivalent thereto; and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence, comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:4, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence, comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-126 of SEQ ID NO:11, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence, comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:4, or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 12 or a sequence equivalent thereto. In another embodiment, an antibody and a fragment thereof are provided that comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 5 or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 10 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:5, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-126 of SEQ ID NO:11, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:5, or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 12 or a sequence equivalent thereto. In another embodiment, an antibody and a fragment thereof are provided that comprise or essentially consist of, or additionally consist of, a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 25-144 of SEQ ID NO: 6 or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, essentially consisting of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO: 10 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence, comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:6, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) variable domain sequence, comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-126 of SEQ ID NO:11, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) variable domain sequence, comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-144 of SEQ ID NO:6, or a sequence equivalent thereto; and/or from a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising, consisting essentially of, or consisting of the sequence of amino acids (aa) 21-126 of SEQ ID NO:12 or a sequence equivalent thereto.

In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 24, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 25, or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI chimeric head peptide. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 1, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 7, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 1, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 8, or a sequence equivalent thereto. In one embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 1, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 9, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 2, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 7, or a sequence equivalent thereto. In one embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:2, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:8, or a sequence equivalent thereto. In one embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:2, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:9, or a sequence equivalent thereto. In one embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:3, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:7, or a sequence equivalent thereto. In one embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:3, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:8, or a sequence equivalent thereto. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:3, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:9, or a sequence equivalent thereto.

In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 26, or a sequence equivalent thereto; and an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 27, or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof binds to the tail region of the DNABII peptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2NTHI. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 4, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 4, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 11, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:4, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:12, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:5, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO:10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 5, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 11, or a sequence equivalent thereto. In yet another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 5, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 12, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 6, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody or fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 6, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 11, or a sequence equivalent thereto. In another embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 6, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence of SEQ ID NO: 12, or a sequence equivalent thereto.

In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:24, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:25, or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI head chimeric peptide). In one embodiment, the antibody or fragment thereof binds to the IhfA5-mIhfB4NTHI head chimeric peptide. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:1, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:7, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:1, or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO: 8 or a sequence equivalent thereto. In another embodiment, an antibody and fragment thereof are provided that comprise or essentially consist of, or additionally consist of, an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO: 1 or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO: 9 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:2, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:7, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:2, or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:8 or a sequence equivalent thereto. In another embodiment, an antibody and fragment thereof are provided that comprise or essentially consist of, or additionally consist of, an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:2 or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:9 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:3, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:7, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:3, or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:8 or a sequence equivalent thereto. In another embodiment, an antibody and fragment thereof are provided that comprise or essentially consist of, or additionally consist of, an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:3 or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-239 of SEQ ID NO:9 or a sequence equivalent thereto.

In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:26, or a sequence equivalent thereto; and an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO:27, or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof binds to the tail region of the DNABII peptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2NTHI. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:4, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO:10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:4, or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO: 11 or a sequence equivalent thereto. In another embodiment, an antibody and fragment thereof are provided that comprise or essentially consist of, or additionally consist of, an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO: 4 or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO: 12 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and an antigen-binding fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:5, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO:10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and an antigen-binding fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:5, or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO: 11 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and an antigen-binding fragment thereof, which comprise or essentially consist of, or additionally consist of, an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO: 5 or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO: 12 or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:6, or a sequence equivalent thereto; and/or an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO:10, or a sequence equivalent thereto. In another embodiment, there are provided an antibody and a fragment thereof, which comprise or essentially consist of, or additionally consist of an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO:6, or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO: 11 or a sequence equivalent thereto. In another embodiment, an antibody and fragment thereof are provided that comprise or essentially consist of, or additionally consist of, an immunoglobulin heavy chain (HC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 25-473 of SEQ ID NO: 6 or a sequence equivalent thereto; and/or from an immunoglobulin light chain (LC) sequence comprising, essentially consisting of, or consisting of the amino acid sequence (aa) 21-233 of SEQ ID NO: 12 or a sequence equivalent thereto.

In another embodiment, there are provided an antibody or fragment thereof, which comprise or consist essentially of or additionally consist of any one, any two, or all three CDR sequences selected from the group comprising the sequences of SEQ ID NOs: 1-3 or 24 or sequences equivalent to each of these; and/or any one, any two, or all three CDR sequences selected from the group comprising the sequences of SEQ ID NOs: 7-9 or 25 or sequences equivalent to each of these. In another embodiment, the antibody or fragment thereof binds to the head region of the DNABII peptide (including, but not limited to: the head region of IHF or HU, the head region of IHFA or IHFB and/or the head chimeric peptide IhfA5-mIhfB4NTHI). In one embodiment, the antibody or fragment thereof binds to the head chimeric peptide IhfA5-mIhfB4NTHI. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody or fragment thereof that comprises or consists essentially of or additionally consists of all three CDR sequences selected from the group comprising the sequences of SEQ ID NO: 1-3 or 24 or the sequences

equivalent to each of the above; and/or from all three CDR sequences selected from the group comprising the sequences of SEQ ID NO: 7-9 or 25 or sequences equivalent to each of the above.

In another embodiment, there are provided an antibody or fragment thereof, which comprise or consist essentially of or additionally consist of any one, any two, or all three CDR sequences selected from the group comprising the sequences of SEQ ID NOs: 4-6 or 26 or sequences equivalent to each of these; and/or any one, any two, or all three CDR sequences selected from the group comprising the sequences of SEQ ID NOs: 10-12 or 27 or sequences equivalent to each of these. In another embodiment, the antibody or fragment thereof binds to the tail region of the DNABII peptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2NTHI. In another embodiment, the fragment is an antigen-binding fragment. In another embodiment, there are provided an antibody or fragment thereof that comprises or essentially consists of or additionally consists of all three CDR sequences selected from the group comprising the sequences of SEQ ID NO: 4-6 or 26 or the sequences,

equivalent to each of the above; and/or from all three CDR sequences selected from the group comprising the sequences of SEQ ID NO: 10-12 or 27 or sequences equivalent to each of the above.

In some embodiments, the antibody or fragment thereof provided in the present application further comprises one or more signal peptides. In one embodiment, the signal peptide comprises or consists essentially of or additionally consists of amino acids (aa) 1-24 of any of the sequences SEQ ID NOs: 1-6, 13, 24 or 26. In another embodiment, the signal peptide comprises or consists essentially of or additionally consists of (aa) 1-20 of any of the sequences SEQ ID NOs: 7-12, 14, 25 and 27. In another embodiment, the signal peptide is located at the amino terminus of the variable region of the light chain. Additionally or alternatively, the same signal peptide or a different signal peptide is located at the amino terminus of the variable region of the heavy chain.

The antibody or fragment thereof provided in the present application may be monospecific or bispecific. In one embodiment, the antibody or fragment thereof is trispecific, tetraspecific, or pentaspecific. Additionally or alternatively, the antibody is selected from the group consisting of an IgA (such as IgA1 or IgA2), IgD, IgE, IgG (such as IgG1, IgG2, IgG3, or IgG4), or IgM antibody. In one embodiment, the antibody further comprises a constant region selected from an IgA constant region (such as an IgA1 constant region or an IgA2 constant region), an IgD constant region, an IgE constant region, an IgG constant region (e.g., an IgG1 constant region, an IgG2 constant region, an IgG3 constant region, or an IgG4 constant region), or an IgM constant region.

In some embodiments, the equivalent amino acid sequence comprises or consists essentially of, or further consists of, a polypeptide having at least about 80% (including from about 80% to 100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical amino acids compared to the reference sequence. Additionally or alternatively, the equivalent amino acid sequence comprises or consists essentially of a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to a polynucleotide complementary to the polynucleotide encoding the reference amino acid sequence. In another embodiment, the equivalent amino acid sequence comprises or consists essentially of, or further consists of, a polypeptide that is at least 80% identical (including from about 80 to 100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) to the original amino acid sequence. In some embodiments, the equivalent amino acid sequence (e.g., the sequences of an antibody or fragment thereof, or any one or more of SEQ ID NOs: 1-14 and 24-26 or fragments thereof, as disclosed herein, including but not limited to: (aa) 25-144 of SEQ ID NO: 13, 24, or 26, (aa) 21-132 of SEQ ID NO: 14 or 25, (aa) 21-126 of SEQ ID NO: 27, (aa) 25-473 of SEQ ID NO: 13, 24, or 26, (aa) 21-239 of SEQ ID NO: 14 or 25, (aa) 21-233 of SEQ ID NO: 27) comprises or consists essentially of or further consists of a polypeptide comprising one or more or all of the CDRs of the original amino acid sequence. Additionally or alternatively, the polypeptide has at least about 80% (including from about 80% to 100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical amino acids compared to the reference sequence.

In some embodiments, the equivalent amino acid sequence, such as that of an antibody, fragment thereof, antigen-binding complementarity determining region (CDR), or CDR-containing polypeptide, has no amino acid differences in the CDRs compared to the CDRs of the original sequence. However, an equivalent amino acid sequence, such as that of an antibody, fragment thereof, CDR, or CDR-containing polypeptide, may have one or more (e.g., without limitation, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acid differences compared to the amino acid sequences of the non-CDR region(s), provided that the three-dimensional orientation of the CDRs and/or CDRs is maintained. In some embodiments, a polypeptide equivalent to an amino acid sequence, such as an antibody sequence, a fragment thereof, a CDR, or a CDR-containing polypeptide, has at least about 80% (including about 80% to 100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical amino acids compared to the original amino acid sequence, provided that the three-dimensional orientation of the CDRs and/or CDRs is maintained.

Non-limiting examples of such non-CDR regions include a framework region (FR), a constant region, an Fc region, a pFc' region, a heavy chain constant domain (CH) (such as CH1, CH2, CH3, or CH4), a light chain constant domain (CL), or a hinge region. In one embodiment, the amino acid differences may be a conservative amino acid substitution and/or do not alter the three-dimensional orientation of the antibody, fragment thereof, CDR, or CDR-containing polypeptide. In another embodiment, the equivalent sequence may include a conservative amino acid substitution within a CDR, such as one or two amino acids at the amino terminus, carboxy terminus, or at both ends of the CDR.

In one embodiment, there are provided one or more CDR regions (such as any 1 or 2 or 3 or 4 or 5 or 6 CDRs) of an antibody or fragment thereof as disclosed herein. In one embodiment, there is provided a set of CDRs comprising or alternatively consisting essentially of or additionally consisting of one or more CDR regions (such as any 1 or 2 or 3 or 4 or 5 or 6 CDRs) of an antibody or fragment thereof as described herein. In one embodiment, there is provided a set of CDRs comprising or alternatively consisting essentially of or additionally consisting of CDRL1, CDRL2 and CDRL3 of the variable region as disclosed herein. In a further embodiment, there is provided a set of CDRs comprising or alternatively consisting essentially of or additionally consisting of CDRH1, CDRH2 and CDRH3 of the variable region as disclosed herein. In another embodiment, a set of CDRs is provided, comprising or alternatively consisting essentially of or additionally consisting of CDRL1, CDRL2 and CDRL3 of a variable region as disclosed herein, and CDRH1, CDRH2, CDRH3 of another variable region as disclosed herein. In some embodiments, the set of CDRs is a paratope. Additionally or alternatively, the set of CDRs specifically binds to a DNABII peptide (such as a head region and/or a tail region, including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, an IhfA5-mIhfB4NTHI chimeric head peptide, an IHF or HU tail region, an IHFA or IHFB tail region and/or an IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, an antibody, fragment thereof, or equivalent of an antibody and fragment thereof is provided, comprising or alternatively essentially consisting of or additionally consisting of any one or more CDR regions as disclosed herein. In yet another embodiment, an antibody, fragment thereof, or equivalent of an antibody and fragment thereof is provided, comprising or alternatively essentially consisting of or additionally consisting of a set of CDRs as disclosed herein.

In some embodiments, the CDRs of SEQ ID NOs: 1-13 are listed in the table below. In some embodiments, CDRH1, any of SEQ ID NOs: 1-6, 13, 24, or 26 comprises or consists essentially of or additionally consists of amino acids (aa) 50-57 of SEQ ID NOs: 1-6, 13, 24, or 26, respectively. In some embodiments, CDRH2, any of SEQ ID NOs: 1-6, 13, 24, or 26 comprises or consists essentially of or additionally consists of amino acids (aa) 75-82 of SEQ ID NOs: 1-6, 13, 24, or 26, respectively. In some embodiments, CDRH3, any of SEQ ID NOs: 1-6, 13, 24, or 26 comprises or consists essentially of or additionally consists of amino acids (aa) 121-133 of SEQ ID NOs: 1-6, 13, 24, or 26, respectively. In some embodiments, CDRL1, any of SEQ ID NOs: 7-9, 14, or 25 comprises or consists essentially of or additionally consists of amino acids (aa) 47-57 of SEQ ID NOs: 7-9, 14, or additionally consists of amino acids (aa) 47-57 of SEQ ID NOs: 7-9, 14, or 25, respectively. In some embodiments, CDRL2, any of SEQ ID NOs: 7-9, 14, or 25 comprises or consists essentially of or additionally consists of amino acids (aa) 75-77 of SEQ ID NOs: 7-9, 14, or 25, respectively. In some embodiments of CDRL3, any of SEQ ID NOs: 7-9, 14, or 25 comprises or consists essentially of or additionally consists of amino acids (aa) 114-122 of SEQ ID NOs: 7-9, 14, or 25, respectively. In some embodiments of CDRL1, any of SEQ ID NOs: 10-12 or 27 comprises or consists essentially of or additionally consists of amino acids (aa) 47-52 of SEQ ID NOs: 10-12 or 27, respectively. In some embodiments of CDRL2, any of SEQ ID NOs: 10-12 or 27 comprises or consists essentially of or additionally consists of amino acids (aa) 70-72 of SEQ ID NOs: 10-12 or 27, respectively. In some embodiments, CDRL3, any of SEQ ID NOs: 10-12 or 27 comprises or consists essentially of or additionally consists of amino acids (aa) 109-116 of SEQ ID NOs: 10-12 or 27, respectively.

SEQ ID NO: CDR1 CDR2 CDR3 Variable region 4 GFTFSRYG ISSGGSYT ERHGGDGYWYFDV 5 GFTFSRYG ISSGGSYT ERHGGDGYWYFDV 6 GFTFSRYG ISSGGSYT ERHGGDGYWYFDV 10 QDISNY YTS QQGNPLRT 11 QDISNY YTS QQGNPLRT 12 QDISNY YTS QQGNPLRT 1 GFTFRTYA IGSDRRHT VGPYDGYYGEFDY 2 GFTFRTYA IGSDRRHT VGPYDGYYGEFDY 3 GFTFRTYA IGSDRRHT VGPYDGYYGEFDY 7 QSLLDSDGKTF LVS WQGTHFPYT 8 QSLLDSDGKTF LVS WQGTHFPYT 9 QSLLDSDGKTF LVS WQGTHFPYT

ND - not defined; SEQ - SEQ ID NO

In some embodiments, the CDRs provided are as shown in the following two tables below.

In some embodiments, an alternative CDR region is provided, which is a CDR as disclosed herein, further comprising 1 additional amino acid, or alternatively 2 amino acids, or alternatively 3 amino acids, or alternatively 4 amino acids, or alternatively 5 amino acids, or alternatively 6 amino acids, or alternatively 7 amino acids, or alternatively 8 amino acids at the amino terminus, or at the carboxy terminus, or at both ends in the corresponding variable region sequence. Additionally or alternatively, an alternative CDR region is provided, which is a CDR as disclosed herein, comprising 1 amino acid, or alternatively 2 amino acids, or alternatively 3 amino acids, or alternatively 4 amino acids, or alternatively 5 amino acids, or alternatively 6 amino acids, or alternatively 7 amino acids, or alternatively 8 amino acids, truncated at the amino terminus, or at the carboxy terminus, or at both ends in the corresponding variable region sequence. For example, CDR1 of SEQ ID NO: 1 may be amino acids 50-57 of SEQ ID NO: 1. However, CDR1 of SEQ ID NO: 1 may also begin with amino acid 42, or 43, or 44, or 45, or 46, or 47, or 48, or 49, or 50, or 51, or 52, or 53, or 54, or 55, or 56, or 57, or 58 of SEQ ID NO: 1. Furthermore, CDR1 of SEQ ID NO: 1 may end with amino acid 49, or 50, or 51, or 52, or 53, or 54, or 55, or 56, or 57, or 58, or 59, or 60, or 61, or 62, or 63, or 64, or 65 of SEQ ID NO: 1, provided that CDR1 ends after the start of SED ID NO: 1. Additionally or alternatively, the CDR comprises about 1, or alternatively about 2, or alternatively about 3, or alternatively about 4, or alternatively about 5, or alternatively about 6, or alternatively about 7, or alternatively about 8, or alternatively about 9, or alternatively about 10, or alternatively about 11, or alternatively about 12, or alternatively about 13, or alternatively about 14, or alternatively about 15 amino acids in length.

In some embodiments, CDR2 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 71-85 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 121-133 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 71-81 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 114-121 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 66-76 of each of SEQ ID NOs: 10-12, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 109-115 of each of SEQ ID NOs: 10-12, respectively.

In some embodiments, CDR1 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 50-57 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 75-82 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 121-122 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR1 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 47-57 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 75-77 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 114-120 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR1 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 47-52 of each of SEQ ID NOs: 10-12, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 70-72 of each of SEQ ID NOs: 10-12, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 109-110 of each of SEQ ID NOs: 10-12, respectively.

In some embodiments, CDR1 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 47-59 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 1-6 comprises or consists essentially of or additionally consists of amino acids 74-83 of each of SEQ ID NOs: 1-6, respectively. In some embodiments, CDR1 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 44-59 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 74-81 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 7-9 comprises or consists essentially of or additionally consists of amino acids 114-122 of each of SEQ ID NOs: 7-9, respectively. In some embodiments, CDR1 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 44-54 of each of SEQ ID NOs: 10-12, respectively. In some embodiments, CDR2 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 79-76 of each of SEQ ID NOs: 10-12, respectively. In some embodiments, CDR3 of any of SEQ ID NOs: 10-12 comprises or consists essentially of or additionally consists of amino acids 109-116 of each of SEQ ID NOs: 10-12, respectively.

In one aspect, one or more variable regions of an antibody or fragment thereof disclosed herein and/or one or more sequences equivalent to the variable regions are provided. In another embodiment, an antibody, fragment thereof, or equivalents of an antibody and fragment thereof are provided, comprising or alternatively consisting essentially of or additionally consisting of any one or any two or more variable regions as disclosed herein and/or one or more sequences equivalent to the sequences of the variable region(s). Additionally or alternatively, one or more variable regions specifically bind to a DNABII peptide (e.g., a head region and/or a tail region, including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, an IhfA5-mIhfB4NTHI chimeric head peptide, an IHF or HU tail region, an IHFA or IHFB tail region, and/or an IhfA3-IhfB2NTHI chimeric tail peptide). In some embodiments, the variable region is selected from the following sequences: amino acids (aa) 25-144 of SEQ ID NOs: 1-6, 13, 24, and 26; (aa) 21-132 of SEQ ID NOs: 7-9, 14, and 25; (aa) 21-126 of SEQ ID NOs: 10-12 or 27; (ak) 24-144 SEQ ID NO: 1-6, 13, 24 and 26; (ak) 20-13 SEQ ID NO: 7-12, 14, 25 and 27; (ak) 20-126 SEQ ID NO: 7-12, 14, 25 and 27.

In another embodiment, the variable region or equivalent region thereof is a variable region as disclosed herein, further comprising 1 additional amino acid, or alternatively 2 amino acids, or alternatively 3 amino acids, or alternatively 4 amino acids, or alternatively 5 amino acids, or alternatively 6 amino acids, or alternatively 7 amino acids, or alternatively 8 amino acids at the amino terminus or at the carboxy terminus, or at both ends of the corresponding sequence provided herein as SEQ ID NO. Additionally or alternatively, the variable region or equivalent region thereof is a variable region as disclosed herein, which is 1 amino acid, or alternatively 2 amino acids, or alternatively 3 amino acids, or alternatively 4 amino acids, or alternatively 5 amino acids, or alternatively 6 amino acids. amino acids, or alternatively 7 amino acids, or alternatively 8 amino acids, truncated at the amino terminus, or at the carboxy terminus, or at both ends of the corresponding sequence provided herein as SEQ ID NO. For example, SEQ ID NO: 1 may be amino acids 50-57 of SEQ ID NO: 1. However, the variable region or equivalent sequence related to the variable region consisting of amino acids 24-144 of SEQ ID NO: 1 may also begin with amino acid 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 or 31 or 32 of SEQ ID NO: 1. Furthermore, the variable region or equivalent sequence related to the variable region consisting of amino acids 24-144 of SEQ ID NO: 1 may end with amino acid 136 or 137 or 138 or 139 or 140 or 141 or 142 or 143 or 144 or 145 or 146 or 147 or 148 or 149 or 150 or 151 or 152 of SEQ ID NO: 1, provided that the variable region ends after the start of SEQ ID NO: 1. Additionally or alternatively, the variable region has a length of from about 90 amino acids to about 200 amino acids, such as a length of about 100 amino acids, or alternatively a length of about 110 amino acids, or alternatively a length of about 120 amino acids, or alternatively a length of about 130 amino acids, or alternatively a length of about 140 amino acids, alternatively a length of about 150 amino acids, or alternatively a length of about 160 amino acids, or alternatively a length of about 170 amino acids, or alternatively a length of about 180 amino acids, or alternatively a length of about 190 amino acids, or alternatively a length of about 200 amino acids.

Additionally or alternatively, the equivalent antibody or fragment thereof comprises one or more (e.g., but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) different amino acids compared to the antibody or fragment thereof in regions other than the variable domain(s) (referred to herein as non-VH regions). Such non-VH regions include, but are not limited to, a constant region, an Fc region, a pFc' region, a heavy chain constant domain (CH) (such as CH1, CH2, CH3, or CH4), a light chain constant domain (CL), or a hinge region. One skilled in the art will appreciate that an antibody, fragment thereof, or equivalent of an antibody and fragment thereof, as disclosed herein, may be further modified in non-VH regions (e.g., to more efficiently couple the heavy chain to the light chain, directly or indirectly conjugate with a detectable marker, a compound that provides purification, or a drug, enhance or reduce complement activation, enhance or reduce antibody-dependent cellular cytotoxicity (ADCC), or increase or decrease activation and recruitment of immune cells) to obtain an additional equivalent.

In some embodiments, the equivalent further comprises up to 50, or alternatively up to 30, or alternatively up to 25, or alternatively up to 20, or alternatively up to 15, or alternatively up to 10, or alternatively up to 5 randomly selected amino acids at either the amino or carboxy terminus, or at both termini. In some embodiments, the equivalent amino acid sequence comprises or consists essentially of or additionally consists of an amino acid sequence shortened at the amino or carboxy terminus, or at both termini, such as by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, or 25 amino acids. Such addition or shortening may not change the three-dimensional orientation of the CDRs and/or the three-dimensional orientation of the antibody, fragment thereof, CDR, or CDR-containing polypeptide.

In some embodiments, the antibody or fragment thereof comprises a signal peptide at the amino terminus of VH and/or at the amino terminus of VL. In one embodiment, the VH signal peptide is different from the VL signal peptide. In another embodiment, the VH signal peptide is the same as the VL signal peptide. In another embodiment, the signal peptide comprises or consists essentially of or additionally consists of amino acids 1-24 of SEQ ID NO: 1. In another embodiment, the signal peptide comprises or consists essentially of or additionally consists of amino acids 1-20 of SEQ ID NO: 7. In some embodiments, the antibody equivalent or fragment thereof comprises a signal peptide that is different from the signal peptide(s) of the parent antibody, provided that the signal peptide of the equivalent directs VH and/or VL to the same site of the cell as the signal peptide(s) of the antibody.

In some embodiments, the equivalent antibody or fragment thereof retains or improves at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%) of one or more functional activities of the antibody or fragment thereof. Such functional activities include, but are not limited to, binding specificity, binding avidity, and/or affinity for a DNABII peptide (such as a head region and/or tail region, including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide), preventing biofilm formation in vivo or in vitro, or disrupting biofilm in vivo or in vitro. Methods for quantifying such functional activity are illustrated in the examples.

In a further aspect, an antibody or fragment thereof is provided that competes for epitope binding with the antibody or fragment thereof disclosed herein. The antibody or fragment thereof may be polyclonal, monoclonal, and/or humanized.

In one embodiment, the antibody is a bispecific antibody or a trispecific, tetraspecific, or pentaspecific antibody. In another embodiment, the antibody is an IgA, IgD, IgE, IgG, or IgM antibody. In another embodiment, the antibodies additionally comprise a constant region selected from an IgA, IgD, IgE, IgG, or IgM constant region. In a specific embodiment, the constant region is an IgG1 constant region. In another embodiment, the present application provides antibodies that compete for epitope binding with the antibody disclosed herein. Such antibodies can be identified using conventional methods, for example, by competitive ELISA.

The antibodies disclosed herein may be polyclonal, monoclonal, or humanized. In one embodiment, the antibodies bind the head region of a DNABII polypeptide, such as HU or IHF (e.g., IhfA and IhfB). In another embodiment, the antibodies bind the tail region of a DNABII polypeptide, such as HU or IHF (e.g., IhfA and IhfB). As noted above, the present application provides antigen-binding fragments. The antigen-binding fragments are any of Fab, F(ab')2, Fab', scFv, or Fv, which can be produced using conventional methods known to those skilled in the art. In some embodiments of the antibody provided herein, the antibody is a soluble Fab. In another embodiment, the present application provides a Fab fragment of the antibody disclosed herein, wherein the antibody specifically binds to the head region of a DNABII peptide (including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment of the invention, DNABII is an IHF or HU peptide. In a specific embodiment, DNABII is an IHF peptide.

As noted above, the present application provides equivalents of antibodies and antigen-binding fragments. An equivalent may include a polypeptide having an amino acid sequence that is at least 80% identical to the sequence of the original polypeptide, or a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to a polynucleotide complementary to the polynucleotide encoding the original polypeptide.

In some embodiments of the antibody disclosed herein, the antibody, fragment, polypeptide, or CDR thereof binds a DNABII protein (including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI head chimeric peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) at a half-maximal effective concentration (EC ₅₀ ) of less than 500 ng/ml, or less than 250 ng/ml, or less than 200 ng/ml, or less than 150 ng/ml, or less than 100 ng/ml, or less than 90 ng/ml, or alternatively less than 80 ng/ml, or alternatively less than 70 ng/ml, or alternatively less than 65 ng/ml, or alternatively less than 60 ng/ml, or alternatively less than 55 ng/ml, or alternatively alternatively less than 50 ng/ml, or alternatively less than 45 ng/ml, or alternatively less than 40 ng/ml, or alternatively less than 35 ng/ml, or alternatively less than 30 ng/ml. In another embodiment, the EC ₅₀ is determined using the ELISA methods set forth in the Examples.

In some embodiments of the antibody disclosed herein, the antibody, fragment, polypeptide, or CDR thereof binds a DNABII protein (including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide) with an equilibrium constant K _D of less than 10 ^-4 M, 10 ^-5 M, 10 ^-6 M, 10 ^-7 M, 10 ^-8 M, 10 ^-9 M, 10 ^-10 M, 10 ^-11 M, or 10 ^-12 M. In one embodiment, the antibody, fragment, polypeptide, or CDR thereof binds a DNABII protein with a K _D of less than 1000 nM, or alternatively less than 900 nM, or alternatively less than 800 nM, or alternatively less than 700 nM, or alternatively less than 600 nM, or alternatively less than 500 nM nM, or alternatively less than 400 nM, or alternatively less than 300 nM, or alternatively less than 200 nM, or alternatively less than 100 nM, or alternatively less than 90 nM, or alternatively less than 80 nM, or alternatively less than 70 nM, or alternatively less than 60 nM, or alternatively less than 50 nM, or alternatively less than 40 nM, or alternatively less than 30 nM, or alternatively less than 20 nM, or alternatively less than 15 nM, or alternatively less than 10 nM, or alternatively less than 9 nM, or alternatively less than 8 nM, or alternatively less than 7 nM, or alternatively less than 6 nM, or alternatively less than 5 nM, or alternatively less than 4 nM, or alternatively less than 3 nM, or alternatively less than 2 nM, or alternatively less than 1 nM. In one embodiment, the K _D is determined using the surface plasmon resonance (SPR) methods provided in the Examples. In some embodiments of the antibody disclosed herein, the antigen-binding site specifically binds to the DNABII protein.

In some embodiments of the antibody disclosed herein, the antibody, fragment, polypeptide, or CDR thereof binds a DNABII protein (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI head chimeric peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) with a K _off of less than 1.0E-02 s ^-1 , or alternatively less than 9.0E-03 s ^-1 , or alternatively less than 8.0E-03 s ^-1 , or alternatively less than 7.0E-03 s ^-1 , or alternatively less than 6.0E-03 s ^-1 , or alternatively less than 5.0E-03 s ^-1 , or alternatively less than 4.0E-03 s ^-1 , or alternatively less than 3.0E-03 s ^-1 , or alternatively less than 2.0E-03 s ^-1 , or alternatively less than 1.0E-03 s ^-1 , or alternatively less than 9.0E-04 s ^-1 , or alternatively less than 8.0E-04 s ^-1 , or alternatively less than 7.0E-04 s ^-1 , or alternatively less than 6.0E -04 s -1 or alternatively less than 5.0E-04 s -1 or alternatively less than 4.0E-04 s ^-1 , or alternatively less than 3.0E-04 s ^-1 , or alternatively less than 2.0E-04 s ^-1 , or alternatively less than 1.0E-04 s ^-1 , or alternatively less than 9.0E-05 s -1 or less than 8.0E-05 s ^-1 , or less than 7.0E-05 s ^-1 , or less than 6.0E-05 s ^-1 , or less than 5.0E -05 s ^-1 , or alternatively less than 4.0E-05 s ^-1 , or alternatively less than 3.0E-05 s ^-1 , or alternatively less than 2.0E-05 s ^-1 , or alternatively less than 1.0E-05 s ^-1 . In one embodiment, K _off is determined using the surface plasmon resonance (SPR) method given in the examples.

In some embodiments of the antibody disclosed herein, the antibody, fragment, polypeptide, or CDR thereof binds a DNABII protein (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI head chimeric peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) with a K _on of less than 9.0E-02 M ^-1 s ^-1 , or alternatively less than 8.0E-02 M ^-1 s ^-1 , or alternatively less than 7.0E-02 M ^-1 s ^-1 , or alternatively less than 6.0E-02 M ^-1 s ^-1 , or alternatively less than 5.0E-02 M ^-1 s ^-1 , or alternatively less than 4.0E-02 M ^-1 s ^-1 , or alternatively less than 3.0E-02 M ^-1 s ^-1 , or alternatively less than 2.0E -02 M ^-1 s ^-1 or alternatively less than 1.0E-02 M ^-1 s ^-1 , or alternatively less than 9.0E-03 M ^-1 s ^-1 , or alternatively less than 8.0E-03 M ^-1 s ^-1 , or alternatively less than 7.0E-03 M ^-1 s ^-1 , or alternatively less than 6.0E-03 M ^-1 s ^-1 , or alternatively less than 5.0E-03 M ^-1 s ^-1 or alternatively less than 4.0E-03 M ^-1 s ^-1 , or alternatively less than 3.0E-03 M ^-1 s ^-1 , or alternatively less than 2.0E-03 M ^-1 s ^-1 , or alternatively less than 1.0E-03 M ^-1 s ^-1 , or less 9.0E-04 M ^-1 s ^-1 , or less than 8.0E-04 M ^-1 s ^-1 , or alternatively less than 7.0E-04 M ^-1 s ^-1 , or alternatively less than 6.0E-04 M ^-1 s ^-1 , or alternatively less than 5.0E-04 M ^-1 s ^-1 , or alternatively less than 4.0E-04 M ^-1 s ^-1 or alternatively less than 3.0E-04 M ^-1 s ^-1 , or alternatively less than 2.0E-04 M ^-1 s ^-1 , or alternatively less than 1.0E-04 M ^-1 s ^-1 , or alternatively less than 9.0 E-05 M ^-1 s ^-1 or alternatively less than 8.0E-05 M ^-1 s ^-1 , or alternatively less than 7.0E-05 M ^-1 s ^-1 , or alternatively less than 6.0E-05 M-1 s-1 or alternatively less than 5.0E-05 M ^-1 s- ¹ , or alternatively less than 4.0E-05 M- ¹ s- ¹ , or alternatively less than 3.0E-05 M ^-1 s ^-1 , or alternatively less than 2.0E-05 M ^-1 s- ¹ or alternatively less than 1.0E-05 M ^-1 s ^-1 . In one embodiment, K _on is determined using the surface plasmon resonance (SPR) method given in the examples.

In some embodiments of the present invention, the association constant K _A for the head chimeric peptide IhfA5-mIhfB4NTHI (in 1/M) is from about 3E+05 to about 2E+08. In another embodiment, K _A is from about 3E+05 to about 1E+08, or alternatively from about 2E+05 to about 1E+08, or alternatively, from about 1E+05 to about 1E+08, or alternatively, from about 1E+06 to about 1E. +08 or alternatively from about 1E+07 to about 1E+08, or alternatively from about 1E+04 to about 1E+09, alternatively from about 1E+05 to about 1E+09, alternatively from about 1E+06 to about 1E+09, alternatively from about 1E+07 to about 1E+09, alternatively from about 1E+08 to about 1E+09, alternatively from about 1E+04 to about 1E+09, or alternatively from about 1E+03 to about 1E+10.

In another embodiment, the dissociation constant K _D for the head chimeric peptide IhfA5-mIhfB4NTHI (in M) is from about 5E-09 to about 3E-06, or alternatively from about 1E-09 to about 1E-06, or alternatively from about 1E-08 to about 1E-05, or alternatively from about 1E-07 to about 1E-05, or alternatively from about 1E-06 to about 1E-05, or alternatively from about 1E-09 to about 1E-08, or alternatively from about 1E-08 to about 1E-07, or alternatively from about 1E-9 to about 1E-08, or alternatively from about 1E-10 to about 1E-09, or alternatively from about 1E-11 to about 1E-10.

In one aspect, the K _A for the IhfA3-IhfB2NTHI tail chimeric peptide (in 1/m) is from about 7E+06 to about 2E+09, or alternatively from about 1E+05 to about 1E+08, or alternatively from about 1E+06 to about 1E+08, or alternatively from about 1E+07 to about 1E+08, or alternatively from about 1E+04 to about 1E+09, alternatively from about 1E+05 to about 1E+09, alternatively from about 1E+06 to about 1E+09, alternatively from about 1E+07 to about 1E+09, alternatively from about 1E+08 to about 1E+09, alternatively from about 1E+04 to about 1E+09, or alternatively from about 1E+03 to about 1E+10, or alternatively from about 1E+03 to about 1E+11, or alternatively from about 1E+03 to about 1E+12, or alternatively from about 1E+09 to about 1E+10, or alternatively from about 1E+10 to about 1E+11, or alternatively from about 1E+11 to about 1E+12.

In another embodiment, the K _D for the IhfA3-IhfB2NTHI tail chimeric peptide (in M) is from about 6E-10 to about 2E-07, or alternatively from about 1E-09 to about 1E-06, or alternatively from about 1E-08 to about 1E-05, or alternatively from about 1E-07 to about 1E-05, or alternatively from about 1E-06 to about 1E-05, or alternatively from about 1E-09 to about 1E-08, or alternatively from about 1E-08 to about 1E-07, or alternatively from about 1E-9 to about 1E-08, or alternatively from about 1E-10 to about 1E-09, or alternatively from about 1E-11 to about 1E-10, or alternatively from about 1E-11 to around 1E-12.

In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds a DNABII protein head region (including, but not limited to, an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI chimeric head peptide) reduces biofilm biomass in vitro by at least about 10%, or alternatively by at least about 15%, or alternatively by at least about 20%, or alternatively by at least about 25%, or alternatively by at least about 30%, or alternatively by at least about 35%, or alternatively by at least about 40%, or alternatively by at least about 45%, or alternatively by at least about 50%, or alternatively by at least about 55%, or alternatively by at least about 60%, or alternatively at least about 65%, or alternatively at least about 70%, or alternatively at least about 75%, or alternatively at least about 80%, or alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 95%. In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds a tail region of a DNABII protein (including, but not limited to: an IHF or HU tail region, an IHFA or IHFB tail region, and/or an IhfA3-IhfB2NTHI tail chimeric peptide) reduces biofilm biomass in vitro by less than about 1%, or alternatively by less than about 2%, or alternatively by less than about 3%, or alternatively by less than about 4%, or alternatively by less than about 5%, or alternatively by less than about 6%, or alternatively by less than about 7%, or alternatively by less than about 8%, or alternatively by less than about 9%, or alternatively by less than about 10%, or alternatively by less than about 12%, or alternatively by less than about 15%, or alternatively by less than about 20%, or alternatively by less than about 25%, or alternatively by less than about 30%, or alternatively by less than about 35%, or alternatively by less than about 40%, or alternatively by less than about 45%, or alternatively by less than about 50%. In one embodiment, such a change in biomass is determined using the methods given in Example 3 or 4.

In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds a head region of a DNABII protein (including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI chimeric head peptide) reduces bacterial load in a subject by at least about 10%, or alternatively by at least about 15%, or alternatively by at least about 20%, or alternatively by at least about 25%, or alternatively by at least about 30%, or alternatively by at least about 35%, or alternatively by at least about 40%, or alternatively by at least about 45%, or alternatively by at least about 50%, or alternatively by at least about 55%%, or alternatively by at least about 60%, or alternatively by at least by at least about 65%, or alternatively by at least about 70%, or alternatively by at least about 75%, or alternatively by at least about 80%, or alternatively by at least about 85%, or alternatively by at least about 90%, or alternatively by at least about 91%, or alternatively by at least about 92%, or alternatively by at least about 93%, or alternatively by at least about 94%, or alternatively by at least about 95%, or alternatively by at least about 96%, or alternatively by at least about 97%, or alternatively by at least about 98% or alternatively by at least about 99%. In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds the tail region of the DNABII protein (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) reduces the bacterial load in a subject by less than about 1%, or alternatively by less than about 2%, or alternatively by less than about 3%, or alternatively by less than about 4%, or alternatively by less than about 5%, or alternatively by less than about 6%, or alternatively by less than about 7%, or alternatively by less than about 8%, or alternatively by less than about 9%, or alternatively by less than about 10%, or alternatively by less than about 12%, or alternatively by less than about 15%, or alternatively by less than about 20%, or alternatively by less by less than about 25%, or alternatively by less than about 30%, or alternatively by less than about 35%, or alternatively by less than about 40%, or alternatively by less than about 45%, or alternatively by less than about 50%. In one embodiment, such a change in bacterial load is determined using the methods provided in the examples.

In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds a DNABII protein head region (including, but not limited to, an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI head chimeric peptide) reduces middle ear occlusion in a subject suffering from otitis media (OM) by at least about 10%, or alternatively by at least about 15%, or alternatively by at least about 20%, or alternatively by at least about 25%, or alternatively by at least about 30%, or alternatively by at least about 35%, or alternatively by at least about 40%, or alternatively by at least about 45%, or alternatively by at least about 50%, or alternatively by at least by at least about 55%, or alternatively by at least about 60%, or alternatively by at least about 65%, or alternatively by at least about 70%, or alternatively by at least about 75%, or alternatively by at least about 80%, or alternatively by at least about 85%, or alternatively by at least about 90%, or alternatively by at least about 91%, or alternatively by at least about 92%, or alternatively by at least about 93%, or alternatively by at least about 94%, or alternatively by at least about 95%, or alternatively by at least about 96%, or alternatively by at least about 97% or alternatively by at least about 98% or alternatively by at least about 99%. In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds the tail region of the DNABII protein (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) reduces middle ear occlusion in a subject suffering from OM by less than about 1%, or alternatively by less than about 2%, or alternatively by less than about 3%, or alternatively by less than about 4%, or alternatively by less than about 5%, or alternatively by less than about 6%, or alternatively by less than about 7%, or alternatively by less than about 8%, or alternatively by less than about 9%, or alternatively by less than about 10%, or alternatively by less than about 12%, or alternatively by less than about 15%, or alternatively by less by less than about 20%, or alternatively by less than about 25%, or alternatively by less than about 30%, or alternatively by less than about 35%, or alternatively by less than about 40%, or alternatively by less than about 45%, or alternatively by less than about 50%. In one embodiment, such a change in middle ear occlusion is determined using the methods given in the examples in an experimental OM model.

In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds a head region of a DNABII protein (including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI head chimeric peptide) reduces the relative mucosal biofilm score and/or the biomass score in a subject with a mucosal biofilm (e.g., having OM) by at least about 10%, or alternatively by at least about 15%, or alternatively by at least about 20%, or alternatively by at least about 25%, or alternatively by at least about 30%, or alternatively by at least about 35%, or alternatively by at least about 40%, or alternatively by at least about 45%, or alternatively by at least about 50%, or alternatively at least about 55%, or alternatively at least about 60%, or alternatively at least about 65%, or alternatively at least about 70%, or alternatively at least about 75%%, or alternatively at least about 80%, or alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 91%, or alternatively at least about 92%, or alternatively at least about 93%, or alternatively at least about 94%, or alternatively at least about 95%, or alternatively at least about 96%, or alternatively at least about 97%, or alternatively at least about 98%, or alternatively at least about 99%. In some embodiments of the antibody disclosed herein, an antibody, fragment, polypeptide, or CDR thereof that binds to a head region of a DNABII protein (including, but not limited to, an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI head chimeric peptide) reduces the relative mucosal biofilm score and/or the biomass score in a subject with a mucosal biofilm (e.g., having an OM) by at least about 0.5, or alternatively by at least about 1, or alternatively by at least about 1.5, or alternatively by at least about 2, or alternatively by at least about 2.5, or alternatively by at least about 3, or alternatively by at least about 3.5, or alternatively by at least about 4. In some embodiments of the antibody disclosed herein, the antibody, its ... a fragment, polypeptide, or CDR that binds the tail region of the DNABII protein (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) reduces the relative mucosal biofilm score and/or the biomass score in a subject with a mucosal biofilm (e.g., having OM) by less than about 1%, or alternatively by less than about 2%, or alternatively by less than about 3%, or alternatively by less than about 4%, or alternatively by less than about 5%, or alternatively by less than about 6%, or alternatively by less than about 7%, or alternatively by less than about 8%, or alternatively by less than about 9%, or alternatively by less than about 10%, or alternatively by less than about 12%, or alternatively by less than about 15%, or alternatively by less by less than about 20%, or alternatively by less than about 25%, or alternatively by less than about 30%, or alternatively by less than about 35%, or alternatively by less than about 40%, or alternatively by less than about 45%, or alternatively by less than about 50%. In some embodiments of the antibody disclosed herein, an antibody, fragment thereof, polypeptide, or CDR that binds the tail region of the DNABII protein (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) reduces the relative mucosal biofilm score and/or the biomass score in a subject with a mucosal biofilm (e.g., having OM) by less than about 0.1, or alternatively by less than about 0.2. or alternatively by less than about 0.3, or alternatively by less than about 0.4, or alternatively by less than about 0.5, or alternatively by less than about 0.6, or alternatively by less than about 0.7, or alternatively by less than about 0.8, or alternatively by less than about 0.9, or alternatively by less than about 1, or alternatively by less than about 1.5, or alternatively by less than about 2, or alternatively by less than about 2.5. In one embodiment, the quantitative indicator is determined using the methods given in the examples.

In some embodiments, the DNABII protein is HU or IHF. In another embodiment, the DNABII protein is IhfA, IhfB, or both. In yet another embodiment, the antibody, fragment, polypeptide, or CDR thereof binds to the head region or head region of the DNABII protein (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the antibody, fragment, polypeptide, or CDR thereof binds the IhfA5-mIhfB4NTHI chimeric head peptide. In another embodiment, the antibody, fragment, polypeptide, or CDR thereof binds the IhfA3-IhfB2NTHI tail chimeric peptide.

In some embodiments of the antibody disclosed herein, the antibody is a soluble Fab.

In some embodiments of the antibody disclosed herein, the HC and LC variable domain sequences are parts of the same polypeptide chain. In some embodiments of the antibody disclosed herein, the HC and LC variable domain sequences are parts of different polypeptide chains.

In some embodiments of the antibody disclosed herein, the antibody is a full-length antibody.

In some embodiments of the antibody disclosed herein, the antibody is chimeric or humanized.

In some embodiments of the antibody disclosed herein, the antibody comprises an Fc domain. In some embodiments of the antibody disclosed herein, the antibody is an antibody from a non-human animal, such as an antibody from a rat, sheep, cow, dog, cat, or rabbit. In some embodiments of the antibody disclosed herein, the antibody is a human or humanized antibody, or is non-immunogenic to humans.

In some embodiments of the antibody disclosed herein, the antibody comprises a framework region of a human antibody. Examples of framework regions that can be combined with LC and HC sequences are known in the art; such examples include SEQ ID NOs: 15-23 or sequences equivalent to each of these.

In other aspects, one or more amino acid residues in a CDR of an antibody of the present invention are replaced with a different amino acid. The substitution may be "conservative" in the sense that it is a substitution within the same group of amino acids. Naturally occurring amino acids can be divided into the following four groups, and conservative substitutions will occur within these groups.

1) Amino acids with basic side chains: lysine, arginine, histidine.

2) Amino acids with acidic side chains: aspartic acid, glutamic acid

3) Amino acids with uncharged polar side chains: asparagine, glutamine, serine, threonine, tyrosine.

4) Amino acids with non-polar side chains: glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan, cysteine.

In another embodiment, one or more amino acid residues are added to or deleted from one or more CDRs of the antibody. Such additions or deletions occur at the N- or C-termini of the CDRs or within the CDRs.

By varying the amino acid sequence of an antibody's CDRs through the addition, deletion, or substitution of amino acids, various effects can be achieved, such as increased binding affinity to the target antigen.

It should be understood that the antibodies of the present invention, although containing such different CDR sequences, still bind the DNABII protein with similar specificity and sensitivity profiles as the original antibodies. This can be verified using binding assays such as ELISA or SPR.

In a further embodiment, the antibodies are both immunodominant and immunoprotective, as determined by appropriate assays and screening.

In another embodiment, the antibodies may be modified by conventional methods that are capable of, for example, increasing the half-life of the antibody, such as by PEGylation, PEG mimeation, polysialylation,

HESylation or glycosylation.

Antibodies and antigen-binding fragments may further comprise a detectable marker or a purification marker.

Antibodies and their derivatives

The present invention also provides an antibody that binds and/or specifically recognizes and binds an isolated polypeptide for use in the methods disclosed herein. The antibody may be any of the various antibodies disclosed herein, and examples of such antibodies include, but are not limited to, a polyclonal antibody, a monoclonal antibody, a chimeric antibody, a human antibody, a veneered antibody, a diabody, a humanized antibody, an antibody derivative, a recombinant humanized antibody, or an equivalent (such as a derivative) or a fragment of each of these compounds. In one embodiment, the fragment comprises or alternatively consists essentially of or additionally consists of a CDR of the antibody. In one embodiment, the antibody is labeled with a detectable label or additionally comprises a detectable label conjugated thereto.

Also provided is a hybridoma cell line that produces the monoclonal antibody disclosed herein. Compositions comprising or alternatively essentially consisting of or, in addition, consisting of one or more of the above-mentioned compounds are further provided herein. Furthermore, provided are polynucleotides that encode the amino acid sequence of antibodies and fragments thereof, as well as methods for the recombinant or chemical synthesis of antibody polypeptides and fragments thereof. Antibody polypeptides can be produced in a eukaryotic or prokaryotic cell or by other methods known in the art and described herein.

Examples of CDR sequences include, but are not limited to, sequences comprising, consisting essentially of, or additionally consisting of, a heavy chain variable region of an antibody or a fragment thereof, or comprising, or alternatively consisting essentially of, a polypeptide encoded by the following polynucleotide sequence, as provided below.

Antibodies can be produced using conventional methods known in the art and well described in the literature. Several methodologies exist for producing polyclonal antibodies. For example, polyclonal antibodies are typically produced by immunizing suitable mammals, such as, but not limited to, chickens, goats, guinea pigs, hamsters, horses, mice, rats, and rabbits. The antigen is introduced into the mammal and induces B lymphocytes to produce immunoglobulins specific to the antigen. Immunoglobulins can be isolated from the mammal's serum.

Variations on this methodology include modification of adjuvants, routes and sites of administration, injection volumes per site, and the number of sites per animal to optimize antibody production and humane animal handling. For example, adjuvants are commonly used to improve or enhance the immune response to antigens. Most adjuvants provide an antigen depot at the injection site, allowing for release of the antigen into draining lymph nodes. Other adjuvants include surfactants, which help concentrate protein antigen molecules over a large surface area, and immunostimulatory molecules. Non-limiting examples of adjuvants for producing polyclonal antibodies include Freund's adjuvants, the Ribi adjuvant system, and Titermax. Polyclonal antibodies can be produced using methods known in the art, some of which are described in U.S. Patents 7,279,559; 7,119,179; 7,060,800; 6,709,659; 6,656,746; 6,322,788; 5,686,073; and 5,670,153.

Monoclonal antibodies can be produced using conventional hybridoma technology methods known in the art and well described in the literature. For example, a hybridoma is produced by fusing a suitable immortalized cell line (e.g., a myeloma cell line such as, but not limited to, Sp2/0, Sp2/0-AG14, NSO, NS1, NS2, AE-1, L.5, P3X63Ag8,653, Sp2 SA3, Sp2 MAI, Sp2 SS1, Sp2 SA5, U397, MIA 144, ACT IV, MOLT4, DA-1, JURKAT, WEHI, K-562, COS, RAJI, NIH 313, HL- 60, MLA 144, NAMAIWA, NEURO 2A, CHO, PerC.6, YB2/O) or the like, or heteromyeloma, fusion products thereof, or any cells or hybrid cells derived therefrom, or any other suitable cell line known in the art (see those available at the following web addresses, e.g., atcc.org, lifetech.com, last accessed November 26, 2007), with antibody-producing cells such as, but not limited to, isolated or cloned spleen, peripheral blood, lymph, tonsil or other immune or B cells, or any other cells expressing heavy or light chain constant region or variable region or framework or CDR sequences, whether endogenous or heterologous nucleic acids or recombinant or endogenous nucleic acids, as well as genomic DNA, cDNA, rDNA, mitochondrial DNA or RNA, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single-, double- or triple-stranded, hybridized nucleic acids, and the like, or any combination thereof. Antibody-producing cells can also be obtained from the peripheral blood or, in specific embodiments, from the spleen or lymph nodes of humans or other suitable animals that have been immunized with the antigen of interest and then screened for the activity of interest. Any other suitable host cell can also be used to express a heterologous or endogenous nucleic acid encoding an antibody, a specific fragment, or a variant thereof according to the present invention. Hybrid cells (hybridomas) or recombinant cells can be isolated using selective culture conditions or other suitable known methods and cloned by limiting dilution, cell sorting, or other known methods.

Other suitable methods for producing or isolating antibodies with the desired specificity may be used, including, but not limited to, methods that select a recombinant antibody from a peptide or protein library (e.g., but not limited to, bacteriophage, ribosome, oligonucleotide, cDNA, and the like libraries, a display library, such as those available from various commercial suppliers such as MorphoSys (Martinsried, Planegg, Del.), BioInvent (Lund, Sweden), Affitech (Oslo, Norway), using methods known in the art. Known methods are described in the patent literature, some of which include U.S. Patent Nos. 4,704,692; 5,723,323; 5,763,192; 5,814,476; 5,817,483; 5,824,514; and 5,976,862. Alternative methods rely on immunization of transgenic animals (e.g., SCID mice, Nguyen et al. (1977) Microbiol. Immunol. 41:901-907 (1997); Sandhu et al. (1996) Crit, Rev. Biotechnol. 16:95-118; Eren et al. (1998) Mumma 93:154-161) that are capable of producing a human antibody repertoire as known in the art and/or as described herein. Such methods include, but are not limited to, ribosome display Wanes et al. (1997) Proc. Natl. Acad. Sci. USA 94:4937-4942; Hanes et al. (1998) Proc. Natl. Acad. Sci. USA 95:14130-14135); single-cell antibody production technologies (e.g., the selected lymphocyte antibody method (SLAM) (U.S. Pat. No. 5,627,052; Wen et al. (1987) J. Immunol 17:887-892; Babcook et al. (1996) Proc. Natl. Acad. Sci. USA 93:7843-7848); gel microdroplets and flow cytometry (Powell et al. (1990) Biotechnol. 8:333-337; One Cell Systems, (Cambridge, Mass.); Gray et al. (1995) J. Imm. KMeth. 182:155-163; and Kenny et al. (1995) Bio. Technol. 13:787-790); B-cell selection (Steenbakkers et al. (1994) Molec. Biol. Reports 19:125-134).

Derivatives of the antibodies of the present invention can also be prepared by delivering a polynucleotide encoding an antibody described herein to a suitable host, for example, to produce transgenic animals or mammals such as goats, cows, horses, sheep, and the like that secrete such antibodies into their milk. These methods are known in the art and are described, for example, in U.S. Patents 5,827,690; 5,849,992; 4,873,316; 5,849,992; 5,994,616; 5,565,362; and 5,304,489.

The term "antibody derivative" includes post-translational modification of the linear polypeptide sequence of an antibody or fragment thereof. For example, U.S. Patent No. 6,602,684 B1 describes a method for producing modified glycol forms of antibodies, including whole antibody molecules, antibody fragments, or fusion proteins, that contain a region equivalent to the Fc region of immunoglobulin exhibiting enhanced Fc-mediated cellular toxicity, and the resulting glycoproteins.

The antibodies described in this application also include derivatives that are modified by covalently attaching any type of molecule to the antibody, such that the covalent attachment does not interfere with the generation of an anti-idiotypic response to the antibody. Antibody derivatives include, but are not limited to, antibodies that have been modified by glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization with known protecting/blocking groups, proteolytic cleavage, binding to a cellular ligand or another protein, etc. In addition, the derivatives may contain one or more atypical amino acids.

Antibody derivatives can also be produced by delivering a polynucleotide described herein to produce transgenic plants and cultured plant cells (e.g., but not limited to, tobacco, corn, and duckweed) that produce such antibodies, specific fragments, or variants in plant parts or in cultured cells derived from these parts. For example, Cramer et al. (1999) Curr. Top. Microbol. Immunol. 240:95-118 and references cited therein describe the production of transgenic tobacco leaves expressing a large number of recombinant proteins, for example, using an inducible promoter. Transgenic corn has been used to express mammalian proteins on an industrial scale, with biological activity equivalent to that of proteins produced in other recombinant systems or purified from natural sources. See, for example, Hood et al. (1999) Adv. Exp. Med. Biol. 464:127–147 and references cited therein. Antibody derivatives, including antibody fragments such as single-chain antibodies (scFv), have also been produced in large quantities from the seeds of transgenic plants, including from tobacco seeds and potato tubers. See, for example, Conrad et al. (1998) Plant Mol. Biol. 38:101–109 and references cited therein. Thus, antibodies can also be produced using transgenic plants according to known methods.

Antibody derivatives can also be produced, for example, by adding exogenous sequences to modify immunogenicity or to reduce, enhance, or modify binding, affinity, association rate, dissociation rate, avidity, specificity, half-life, or any other suitable property. Typically, portions or full-length CDR sequences of non-human or human origin are retained, while the variable and constant region sequences of non-human origin are replaced with human or other amino acids, or variable or constant regions of other isotypes.

In general, the CDR residues are directly and most significantly involved in influencing antigen binding. Humanization or engineering of antibodies can be accomplished using any known method, such as, but not limited to, those described in U.S. Patent Nos. 5,723,323; 5,976,862; 5,824,514; 5,817,483; 5,814,476; 5,763,192; 5,723,323; 5,766,886; 5,714,352; 6,204,023; 6,180,370; 5,693,762; 5,530,101; 5,585,089; 5,225,539; and 4,816,567.

Chimeric, humanized, or primatized antibodies of the present invention can be prepared based on the sequence of a reference monoclonal antibody obtained using standard molecular biology techniques. DNA encoding heavy and light chain immunoglobulins can be obtained from the hybridoma of interest and engineered to contain non-reference (e.g., human) immunoglobulin sequences using standard molecular biology techniques. For example, to create a chimeric antibody, murine variable regions can be linked to human constant regions using methods known in the art (U.S. Patent No. 4,816,567). To create a humanized antibody, the mouse CDR regions can be inserted into the framework region of a human antibody using methods known in the art (U.S. Pat. No. 5,225,539 and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370). Similarly, to create a primatized antibody, the mouse CDR regions can be inserted into the framework region of a primate antibody using methods known in the art (WO 93/02108 and WO 99/55369).

Methods for producing partially or fully human antibodies are known in the art, and any such methods can be used. According to one embodiment, fully human antibody sequences are produced in transgenic mice engineered to express the heavy and light chain genes of human antibodies. Several lines of such transgenic mice have been generated, capable of producing different classes of antibodies. B cells from transgenic mice that produce the antibody of interest can be combined to generate hybridoma cell lines for continuous production of the antibody of interest (See, e.g., Russell et al. (2000) Infection and Immunity April 2000:1820–1826; Gallo et al. (2000) European J. of Immun. 30:534–540; Green (1999) J. of Immun. Methods 231:11–23; Yang et al. (1999A) J. of Leukocyte Biology 66:401–410; Yang (1999B) Cancer Research 59(6):1236–1243; Jakobovits (1998) Advanced Drug Reviews 31:33–42; Green and Jakobovits (1998) J. Exp. Med. 188(3):483-495; Jakobovits (1998) Exp. Opin. Invest. Drugs 7(4):607-614; Tsuda et al. (1997) Genomics 42:413-421; Sherman-Gold (1997) Genetic Engineering News 17(14); Mendez et al. (1997) Nature Genetics 15:146-156; Jakobovits (1996) Weir's Handbook of Experimental Immunology, The Integrated Immune System Vol. IV, 194.1-194.7; Jakobovits (1995) Current Opinion in Biotechnology 6:561-566; Mendez et al. (1995) Genomics 26:294-307; Jakobovits (1994) Current Biology 4(8):761-763; Arbones et al. (1994) Immunity 1(4):247–260; Jakobovits (1993) Nature 362(6417):255-258; Jakobovits et al. (1993) Proc. Natl. Acad. Sci. USA 90(6):2551-2555; and Pat. US No. 6,075,181).

The antibodies disclosed herein can also be modified to create chimeric antibodies. Chimeric antibodies are antibodies in which different heavy and light chain domains of the antibody are encoded by DNA derived from more than one species. See, for example, U.S. Patent No. 4,816,567.

Alternatively, the antibodies described herein can also be modified to create veneered antibodies. Veneered antibodies are antibodies in which the external amino acid residues of antibodies from one mammalian species are intentionally replaced, or "veneered," with residues from antibodies from a second mammalian species so that the antibodies of the first species will not be immunogenic for the second species, thereby reducing the immunogenicity of the antibody. Since the antigenicity of a protein depends primarily on the nature of its surface, the immunogenicity of an antibody can be reduced by replacing exposed residues that differ from those typically found in other mammalian species. This intentional substitution of external residues should have little or no effect on internal domains or on interdomain contacts. Thus, ligand-binding properties should not be affected by changes that are limited to residues of the framework region of the variable region. This process is called "veneering" because only the outer surface or "shell" of the antibody is changed, while the supporting residues remain intact.

The veneering procedure utilizes available human antibody variable domain sequence data compiled by Kabat et al. (1987) Sequences of Proteins of Immunological Interest, 4th ed., Bethesda, Md., National Institutes of Health, updates of this database, and other available U.S. and international database materials (both nucleic acid and protein). Non-limiting examples of methods used to produce veneered antibodies include EP 519,596; U.S. Patent No. 6,797,492; and Padlan et al. (1991) Mol. Immunol. 28(4-5):489–498.

The term "antibody derivative" also includes "diabodies," which are small antibody fragments with two antigen-binding sites, the fragments comprising a heavy-chain variable domain (VH) linked to a light-chain variable domain (VL) in the same polypeptide chain. (See, e.g., EP 404 097, WO 93/11161, and Hollinger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444–6448.) By using a linker that is too short to allow pairing between the two domains in the same chain, the domains are forced to pair with complementary domains of another chain and create two antigen-binding sites. (See also U.S. Patent No. 6,632,926 to Chen et al., which describes antibody variants that have one or more amino acids inserted into the hypervariable region of the parent antibody and a binding affinity for a target antigen that is at least about two-fold higher than the binding affinity of the parent antibody for the antigen.)

The term "antibody derivative" further includes engineered antibody molecules, fragments, and individual domains such as scFvs, dAbs, nanobodies, minibodies, unitels, and affibodies (Hudson (2005) Nature Biotech 23(9):1126-36; Published U.S. Patent Application No. 2006/0211088; Published International Application WO 2007/059782; U.S. Patent No. 5,831,012).

The term "antibody derivative" further includes "linear antibodies." The procedure for producing linear antibodies is known in the art and is described by Zapata et al. (1995) Protein Eng. 8(10):1057–1062. Briefly, these antibodies contain two tandem Ed segments (VH-CH1-VH-CH1) that form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

The antibodies described in this application can be isolated and purified from recombinant cell cultures by known methods, including, but not limited to, protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion-exchange or cation-exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxyapatite chromatography, and lectin chromatography. High-performance liquid chromatography ("HPLC") can also be used for purification.

The antibodies of the present invention include naturally occurring purified products, products obtained by chemical synthetic techniques, and products produced by recombinant methods in eukaryotic host cells, including, for example, yeast, higher plant, insect, and mammalian cells, or alternatively in prokaryotic host cells, as described above. A number of antibody production systems are described in Birch & Radner (2006) Adv. Drug Delivery Rev. K58: 671-685.

If a test antibody binds to a protein or polypeptide, then the test antibody and the antibodies provided herein are equivalent. It can also be determined without undue experimentation whether an antibody has the same specificity as an antibody disclosed herein by determining whether the test antibody interferes with the binding of the antibody disclosed herein to a protein or polypeptide with which the antibody normally reacts. If the test antibody competes with the antibody described herein, as evidenced by a decrease in binding by the monoclonal antibody described herein, then it is likely that the two antibodies bind to the same or a closely related epitope. Alternatively, the antibody described herein can be pre-incubated with a protein with which it normally reacts and the ability of the test antibody to bind the antigen is inhibited. If such activity is inhibited, then in all likelihood it has the same or closely related epitope specificity as the antibody described in the present application.

The term "antibody" also encompasses antibodies of all immunoglobulin isotypes and subclasses. Specific isotypes of a monoclonal antibody can be obtained either directly by selection from the initially fused cells or secondarily obtained from a parental hybridoma secreting a monoclonal antibody of a different isotype using clonal selection to isolate class-switched variants using the procedure described by Steplewski et al. (1985) Proc. Natl. Acad. Sci. USA 82:8653 or Spira et al. (1984) J. Immunol. Methods 74:307. Alternatively, recombinant DNA techniques can be used.

The isolation of other monoclonal antibodies with the specificity of the monoclonal antibodies described in this application can also be accomplished by one skilled in the art by producing anti-idiotypic antibodies. Herlyn et al. (1986) Science 232:100. An anti-idiotypic antibody is an antibody that recognizes unique determinants present on the monoclonal antibody of interest.

In some embodiments disclosed herein, a detectably or therapeutically labeled antibody will be useful. Suitable labels are described above. Methods for conjugating antibodies to such labels are known in the art. For illustrative purposes only, antibodies can be labeled with a detectable label, such as a radioactive atom, chromophore, fluorophore, etc. Such labeled antibodies can be used for diagnostic assays either in vivo or in an isolated test sample.

Binding antibodies to low-molecular-weight haptens can increase the sensitivity of the antibody in the assay. Haptens can then be specifically detected using a second reaction. For example, commonly used haptens include biotin, which reacts with avidin, or dinitrophenol, pyridoxal, and fluorescein, which can react with specific antibodies against the hapten. See above, Harlow and Lane (1988).

The variable region of the antibodies of the present invention can be modified by mutating amino acid residues in the VH and/or VL regions of CDR 1, CDR 2 and/or CDR 3 to improve one or more binding properties (e.g., affinity) of the antibody. Mutations can be introduced using site-directed mutagenesis or PCR-mediated mutagenesis, and the effect on antibody binding or another functional property of interest can be assessed in appropriate in vitro or in vivo assays. In some embodiments, conservative modifications are introduced and typically no more than one, two, three, four or five residues in the CDR region are changed. Mutations can be amino acid substitutions, additions or deletions.

To reduce immunogenicity, modifications can be made to the framework regions of antibodies, for example by "back-mutating" one or more framework residues in the corresponding germline sequence.

Furthermore, the antibodies described herein can be engineered to include modifications in the Fc region to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or antigen-dependent cellular cytotoxicity. Such modifications include, but are not limited to, changes in the number of cysteine residues in the hinge region to facilitate assembly of the light and heavy chains or to increase or decrease antibody stability (U.S. Patent No. 5,677,425) and amino acid mutations in the Fc hinge region to decrease the biological half-life of the antibody (U.S. Patent No. 6,165,745).

Furthermore, the antibodies described herein can be chemically modified. Glycosylation of an antibody can be altered, for example, by modifying one or more glycosylation sites in the antibody sequence to increase the affinity of the antibody for the antigen (U.S. Patent Nos. 5,714,350 and 6,350,861). Alternatively, to enhance antibody-dependent cell-mediated cytotoxicity, a hypofucosylated antibody having a reduced number of fucosyl residues or an antibody having an increased number of cleaved GlcNac bisector structures can be produced by expressing the antibody in a host cell with an altered glycosylation machinery (Shields, RL et al. (2002) J. Biol. Chem. 277:26733-26740; Umana et al. 16:95-118; Eren et al. Biotech. 17:176-180).

The antibodies disclosed herein can be PEGylated to increase biological half-life by reacting the antibody or antibody fragment with polyethylene glycol (PEG) or a reactive ester or aldehyde derivative of PEG under conditions in which one or more PEG groups are attached to the antibody or antibody fragment. PEGylation of the antibody can be accomplished by an acylation reaction or an alkylation reaction with a reactive PEG molecule (or a similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" refers to any of the forms of PEG that have been used to derivatize other proteins, such as mono (C1-C10) alkoxy- or aryloxy-polyethylene glycol or polyethyleneglycol-maleimide. The antibody to be PEGylated can be an aglycosylated antibody. Methods for PEGylation of proteins are known in the art and can be applied to the antibodies described in the present application (EP 0 154 316 and EP 0 401 384).

Furthermore, antibodies can be chemically modified by conjugating or fusing the antigen-binding region of the antibody to a serum protein, such as human serum albumin, to increase the half-life of the resulting molecule. This approach is described, for example, in EP 0 322 094 and EP 0 486 525.

The antibodies or fragments thereof of the present invention can be conjugated to a diagnostic agent and used for diagnostic purposes, such as monitoring the development or progression of a disease and determining the effectiveness of a treatment regimen. Examples of diagnostic agents include enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron-emitting metals using various positron emission tomographs, and non-radioactive paramagnetic metal ions. The detectable substance can be linked or conjugated either directly to the antibody or fragment thereof or indirectly, through a linker, using methods known in the art. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase. Examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin. An example of a luminescent material includes luminol. Examples of bioluminescent materials include luciferase, luciferin, and aequorin. Examples of suitable radioactive molecules include 125I, 131I, indium-111, lutetium-171, bismuth-212, bismuth-213, astatine-211, copper-62, copper-64, copper-67, yttrium-90, iodine-125, iodine-131, phosphorus-32, phosphorus-33, scandium-47, silver-111, gallium-67, praseodymium-142, samarium-153, terbium-161, dysprosium-166, holmium-166, rhenium-186, rhenium-188, rhenium-189, lead-212, radium-223, actinium-225, iron-59, selenium-75, arsenic-77, strontium-89, molybdenum-99, rhodium-1105, palladium-109, praseodymium-143, promethium-149, erbium-169, iridium-194, gold-198, gold-199, and lead-211. Monoclonal antibodies can be indirectly conjugated to radioactive metal ions through the use of bifunctional chelating agents that are covalently linked to the antibodies. Chelating agents can be attached via amides (Meares et al. (1984) Anal. Biochem. 142:68–78) and sulfhydryl groups (Koyama (1994) Chem. Abstr. 120:217–262) to amino acid residues and carbohydrate groups (Rodwell et al. (1986) PNAS USA 83:2632–2636; Quadri et al. (1993) Nucl. Med. Biol. 20:559–570).

In addition, the antibodies or fragments thereof of the present invention can be conjugated to a therapeutic agent. Suitable therapeutic agents include the toxins taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoside, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracinatedione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol and puromycin, antimetabolites (such as methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5-fluorouracil, decarbazine, hydroxyurea, asparaginase, gemcitabine, cladribine), alkylating agents (such as mechlorethamine, thioepa, chloramhucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, cisplatin and other platinum derivatives such as carboplatin), antibiotics (such as dactinomycin (formerly actinomycin), bleomycin, daunorubicin (formerly daunomycin), doxorubicin, idarubicin, mithramycin, mitomycin, mitoxantrone, plicamycin, anthramycin (AMC)), diphtheria toxin and related molecules (such as diphtheria chain A and its active fragments and fusion molecules), ricin toxin (such as ricin A or deglycosylated ricin A chain toxin), cholera toxin, Shiga-like toxin (SLT-I, SLT-II, SLT-IIV), LT toxin, C3 toxin, Shiga toxin, pertussis toxin, tetanus toxin, soybean protease inhibitor Bowman-Birk, Pseudomonas exotoxin, alorin, saporin, modeccin, gelonin, abrin A chain, modessin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolacca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrietocin, phenomycin, enomycin, and mixed toxins.

Additional suitable molecules for conjugation include ribonuclease (RNase), DNase I, antisense nucleic acid, inhibitory RNA molecule such as an siRNA molecule, immunostimulatory nucleic acid, aptamers, ribozymes, triplex-forming molecules, and external targeting sequences. Aptamers are small nucleic acids, 15 to 50 bases in length, that fold into specific secondary and tertiary structures such as stem-loop or G-quadruplexes and can bind small molecules such as ATP (U.S. Patent No. 5,631,146) and theophylline (U.S. Patent No. 5,580,737), as well as large molecules such as reverse transcriptase (U.S. Patent No. 5,786,462) and thrombin (U.S. Patent No. 5,543,293). Ribozymes are nucleic acid molecules that are capable of catalyzing a chemical reaction both intramolecularly and intermolecularly. Ribozymes typically cleave nucleic acid substrates by recognizing and binding the target substrate, followed by cleavage. Nucleic acid molecules with triplex-forming properties can interact with double- or single-stranded nucleic acids, forming a triplex in which the three DNA strands form a complex dependent on both Watson-Crick and Hoogsteen base pairing. Triplex molecules can bind to target regions with high affinity and specificity.

Functional nucleic acid molecules can act as effectors, inhibitors, modulators, and stimulators of a specific activity possessed by a target molecule, or functional nucleic acid molecules can have a de novo activity independent of any other molecules.

Therapeutic agents can be linked to the antibody directly or indirectly using any of a number of available methods. For example, the agent can be attached to the hinge region of the reduced antibody component through disulfide bond formation using crosslinking agents such as N-succinyl 3-(2-pyridyldithio)propionate (SPDP), or through a carbohydrate moiety in the Fc region of the antibody (Yu et al. 1994 Int. J. Cancer 56: 244; Upeslacis et al., “Modification of Antibodies by Chemical Methods,” in Monoclonal antibodies: principles and applications, Birch et al. (eds.), pages 187-230 (Wiley-Liss, Inc. 1995); Price, “Production and Characterization of Synthetic Peptide-Derived Antibodies,” in Monoclonal antibodies: Production, engineering and clinical application, Ritter et al. (eds.), pages 60-84 (Cambridge University Press 1995)).

Methods for conjugating therapeutic agents with antibodies are well known (Amon et al. “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy,” in Monoclonal Antibodies And Cancer Therapy; Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al. “Antibodies For Drug Delivery,” in Controlled Drug Delivery (2nd Ed.); Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); The Therapeutic Use of Radiolabeled Antibody in Cancer Therapy,” in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al. “The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates,” (1982) Immunol. Rev. 62:119-58).

The antibodies disclosed herein, or their antigen-binding portions, can be linked to another functional molecule, such as another antibody or a ligand for a receptor, to form a bispecific or multispecific molecule that binds to at least two or more different binding sites or target molecules. Linking of an antibody to one or more other binding molecules, such as another antibody, antibody fragment, peptide, or binding mimetic, can be accomplished, for example, by chemical linkage, genetic fusion, or non-covalent association. Multispecific molecules can further comprise a third binding specificity in addition to the specificity for the first and second target epitopes.

Bispecific and multispecific molecules can be produced using methods known in the art. For example, each linker of a highly specific molecule can be produced separately and then conjugated to another linker. When the linkers are proteins or peptides, various coupling or cross-linking agents can be used for covalent conjugation. Examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-S-acetylthioacetate (SATA), 5,5'-dithiobis(2-nitroberyzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), and sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-I-carboxylate (sulfo-SMCC) (Karpovsky et al. (1984) J. Exp. Med. 160:1686; Liu et al. (1985) Proc. Natl. Acad. Sci. USA 82:8648). When the binding molecules are antibodies, they can be conjugated via a sulfhydryl bond between the C-terminal hinge regions of the two heavy chains.

The antibodies or fragments thereof of the present invention can be linked to a fragment that is toxic to the cell to which the antibody binds, forming "depleting" antibodies. These antibodies are particularly useful in cases where depletion of NK cells is desired.

The antibodies described herein can also be attached to solid supports, which is particularly useful for immunoassays or target antigen purification. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene.

Antibodies can also be bound to a variety of carriers. Thus, the present invention also provides compositions comprising antibodies and another substance, either active or inert. Examples of well-known carriers include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylase, natural and modified cellulose, polyacrylamide, agarose, and magnetite. The carrier can be either soluble or insoluble for the purposes disclosed herein. Those skilled in the art will recognize other suitable carriers for binding monoclonal antibodies or will be able to determine such through routine experimentation.

In some embodiments, the invention relates to an antibody or antigen-binding fragment that specifically recognizes or binds the head or tail domain of the DNABII protein or a fragment thereof, the tail fragment, or the head fragment. The DNABII protein or fragment thereof may be an IHF or HU polypeptide.

Functional analysis with antibodies

The antibodies disclosed herein can be used to purify the polypeptides disclosed herein and to identify biologically equivalent polypeptides and/or polynucleotides. They can also be used to identify agents that modify the function of the polypeptides described herein. These antibodies include polyclonal antisera, monoclonal antibodies, and various reagents derived therefrom, which are known to those skilled in the art and described above.

Antibodies that neutralize the activity of proteins encoded by the identified genes can also be used in vivo and in vitro to demonstrate function by adding such neutralizing antibodies to in vivo and in vitro testing systems. They can also be used as pharmaceutical agents to modulate the activity of the polypeptides disclosed in this application.

Various antibody preparations can also be used in analytical methods, such as ELISA assays or Western blots, to demonstrate the expression of proteins encoded by identified genes in test cells in vitro or in vivo. Fragments of such proteins, formed during protease degradation during metabolism, can also be identified using appropriate polyclonal antisera with samples obtained from experimental specimens.

The antibodies disclosed herein can be used for vaccination or booster vaccination, alone or in combination with peptide- or protein-based vaccines or dendritic cell-based vaccines.

Diagnostic and therapeutic methods

A method is provided for preventing, inhibiting, or competing for the binding of a DNABII polypeptide or protein to microbial DNA by contacting the DNABII polypeptide or protein or microbial DNA with an effective amount of one or more agents described above, for example, with an antibody or an antigen-binding fragment thereof, a polypeptide or CDR, as disclosed herein, thereby preventing, inhibiting, or competing for the binding of the DNABII protein or polypeptide to microbial DNA. The DNABII polypeptide may be an IHF or HU peptide. In one embodiment, the antibody or antigen-binding fragment thereof selectively binds to the head region of the DNABII polypeptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 24 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 25 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or consisting essentially of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 1, 2 or 3 or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or consisting essentially of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 7, 8 or 9 or sequences equivalent to each of said sequences. In another embodiment, the contacting is carried out in vivo or in vitro.

In another embodiment, the antibody or antigen-binding fragment thereof for use in the method disclosed herein comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising a sequence selected from the group of sequences of SEQ ID NO: 13, 24 or 26, or sequences equivalent to each of these, and/or an immunoglobulin light chain (LC) variable domain sequence comprising a sequence selected from the group of sequences of SEQ ID NO: 14, 25 or 27, or sequences equivalent to each of these.

In another embodiment, one or more of the DNABII polypeptide and/or microbial DNA and/or antibody, or antigen-binding fragment thereof, and/or polypeptide or CDR, as disclosed herein, are labeled with a detectable, such as a radioisotopic label, or luminescent molecules that will emit a signal upon close contact with each other. The contacting can be carried out in vitro or in vivo. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or destruction, for example, using one or more DNABII polypeptides and/or microbial DNA and/or antibody, or antigen-binding fragment thereof, and/or polypeptide or CDR, as disclosed herein. In one aspect, diagnostic methods comprise using an antibody or antigen-binding fragment as disclosed herein, which in one embodiment specifically binds to the tail region or fragment of a DNABII polypeptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or essentially consists of or additionally consists of a heavy chain (HC) variable domain sequence of an immunoglobulin comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of SEQ ID NOs: 4, 5 or 6, or sequences equivalent to each of these, and/or a light chain (LC) variable domain sequence of an immunoglobulin comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of SEQ ID NOs: 10, 11 or 12, or sequences equivalent to each of these.

In another embodiment, a method for disrupting a microbial biofilm is provided by contacting the biofilm with an effective amount of one or more agents as described above, such as an antibody or an antigen-binding fragment thereof, a polypeptide, or a CDR, thereby disrupting the microbial biofilm. In one embodiment, the antibody or antigen-binding fragment thereof selectively binds to the head region of a DNABII polypeptide (including, but not limited to, the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). The DNABII polypeptide may be an IHF or HU peptide. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 24 or or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 25 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or consisting essentially of or additionally consisting of an amino acid sequence selected from the group of SEQ ID NOs: 1, 2 or 3, or sequences equivalent to each of these, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or consisting essentially of or additionally consisting of an amino acid sequence selected from the group of SEQ ID NOs: 7, 8 or 9, or sequences equivalent to each of these. In yet another embodiment, the contacting is carried out in vivo or in vitro.

In one embodiment, the microbial biofilm is produced by a microorganism that exports a DNABII polypeptide. The DNABII polypeptide may be an IHF or HU peptide. In another embodiment, one or more of the antibodies, antibody fragments, DNABII polypeptide, and microbial DNA are labeled with a detectable label, such as a radioisotope or luminescent molecules that emit a signal upon close contact with one another. The contacting can be carried out in vitro or in vivo. In one embodiment, the agent is one or more antibodies and/or antigen-binding fragments that are the same or different from one another. In some embodiments, such antibodies or antigen-binding fragments are administered alone or in combination with one another, or with an agent other than an antibody, or with an additional pharmaceutically effective agent, alone or in combination with a pharmaceutically acceptable carrier. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or destruction. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as described herein, which in one embodiment specifically binds to the tail region or tail fragment of a DNABII polypeptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

Also provided are methods for preventing the formation or destruction of a biofilm on a surface, comprising or consisting essentially of or further consisting of treating a surface susceptible to or containing a biofilm with an effective amount of one or more agents selected from an antibody or antigen-binding fragment thereof, a polypeptide, or a CDR as described herein, wherein the antibody or antigen-binding fragment thereof binds the head region of a DNABII peptide (including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI chimeric head peptide). The DNABII polypeptide may be an IHF or HU peptide. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or essentially consists of or additionally consists of a heavy chain (HC) variable domain sequence of an immunoglobulin comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of SEQ ID NOs: 4, 5 or 6, or sequences equivalent to each of these, and/or a light chain (LC) variable domain sequence of an immunoglobulin comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of SEQ ID NOs: 10, 11 or 12, or sequences equivalent to each of these.

In one embodiment, the antibody or antigen-binding fragment thereof comprises a detectable label. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or disruption. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as disclosed herein, which in one aspect specifically binds to the tail region or fragment of a DNABII polypeptide (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or essentially consists of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 4, 5 or 6, or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 10, 11 or 12, or sequences equivalent to each of said sequences.

Furthermore, the present invention provides methods for detecting a biofilm in a subject, comprising or essentially consisting of or consisting of administering to the subject an effective amount of one or more antibodies or fragments thereof, a polypeptide or CDR disclosed herein. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as disclosed herein, which in one aspect specifically binds to the tail region or fragment of a DNABII polypeptide (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI tail chimeric peptide). The DNABII polypeptide may be an IHF or HU peptide. In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 4, 5 or 6, or sequences equivalent to each of said sequences, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 10, 11 or 12, or sequences equivalent to each of said sequences.

Methods for preventing or disrupting a biofilm in a subject are provided. The methods comprise or essentially consist of or consist of administering to the subject an antibody or fragment thereof as disclosed herein that binds to the head region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, methods for preventing or disrupting a biofilm in a subject are provided, comprising or alternatively consisting of or further consisting of administering to the subject an effective amount of one or more antibodies, fragments thereof, polypeptides, or CDRs as described herein, and/or an effective amount of one or more polynucleotides or vectors encoding the antibody, fragment thereof, polypeptide, or CDR. The DNABII peptide may be an IHF or HU peptide. In one embodiment, the antibody or antigen-binding fragment thereof selectively binds to the head region of a DNABII polypeptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 24 or a sequence equivalent thereto, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 25 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or essentially consists of or additionally consists of: an immunoglobulin heavy chain (HC) variable domain sequence comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences SEQ ID NO: 1, 2 or 3, or sequences equivalent to each of said, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences SEQ ID NO: 7, 8 or 9, or sequences equivalent to each of said. In another embodiment, the contact is carried out in vivo or in vitro. The antibody or antigen-binding fragment thereof can be labeled with a detectable label. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or destruction. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as disclosed herein, which in one aspect specifically binds to the tail region or fragment of a DNABII polypeptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, and/or the tail chimeric peptide IhfA3-IhfB2NTHI). In another aspect, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

Provided are methods for treating a condition characterized by biofilm formation in a subject, comprising or consisting essentially of or consisting of administering to the subject an antibody or fragment thereof, as disclosed herein, that binds to the head region of a DNABII polypeptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, the methods for preventing or treating a condition characterized by biofilm formation in a subject are provided by administering to the subject an effective amount of one or more antibodies, fragments thereof, polypeptides, or CDRs, as disclosed herein, and/or an effective amount of one or more polynucleotides or vectors encoding the antibody, fragment thereof, polypeptide, or CDR. In another embodiment, methods are provided for inhibiting, preventing, or treating a microbial infection that causes biofilm formation in a subject, comprising or alternatively consisting of or further consisting of administering to the subject an effective amount of one or more compounds selected from an antibody, a fragment thereof, a polypeptide, or a CDR, as described herein, and/or an effective amount of one or more polynucleotides or vectors encoding the antibody, a fragment thereof, a polypeptide, or a CDR. The DNABII polypeptide may be an IHF or HU peptide. In one embodiment, the antibody or antigen-binding fragment thereof selectively binds to the head region of the DNABII polypeptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 24 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 25 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or essentially consists of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 1, 2 or 3, or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or essentially consisting of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 7, 8 or 9, or sequences equivalent to each of said sequences. In another embodiment, the contact is carried out in vivo or in vitro. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or destruction. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as described herein, which in one aspect specifically binds to the tail region or tail fragment of a DNABII polypeptide (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

In the above methods, the biofilm originates from gram-negative or gram-positive biofilm-producing bacteria. Non-limiting examples of conditions include chronic non-healing wounds, including venous ulcers and diabetic foot ulcers, ear infections, sinus infections, urinary tract infections, gastrointestinal diseases, pulmonary infections, respiratory tract infections, cystic fibrosis, chronic obstructive pulmonary disease, catheter-related infections, implanted device infections, implanted prosthetic infections, osteomyelitis, cellulitis, abscesses, and periodontal disease.

In some embodiments of the method disclosed herein, administering one or more antibodies, fragments, polypeptides, or CDRs thereof reduces the level of one or more proinflammatory cytokines in the subject. Non-limiting examples of proinflammatory cytokines include: IL-1β, IL6, IL8, IL12p70, IL17A, interferon (IFN), and tumor necrosis factor (TNF). Additionally or alternatively, administering an effective amount of one or more antibodies, fragments, polypeptides, or CDRs thereof increases the level of one or more anti-inflammatory cytokines in the subject. In one embodiment, anti-inflammatory cytokines include, but are not limited to, IL-10, IL-13, IL-1ra, IL-4, IL-11, and transforming growth factor-β (TGF-β).

Also provided are methods for inducing a proinflammatory response or treating a condition mediated by an attenuated inflammatory response in a subject in need thereof, wherein the method comprises or alternatively essentially consists of or additionally consists of administering to the subject an effective amount of an antibody that binds to the tail region of a DNABII polypeptide (including, but not limited to, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide). The DNABII polypeptide may be an IHF or HU peptide. These methods can be combined with diagnostic methods for detecting and/or monitoring the release or levels of cytokines in the tissues of the subject. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as described herein, which in one aspect specifically binds to the tail region or tail fragment of a DNABII polypeptide (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label. Non-limiting examples of cytokines include, for example, IL1beta, IL6, IL8, IL12p70, IL10, IL13, and IFN.

Also provided are methods for inhibiting a proinflammatory response or treating a condition mediated by an enhanced inflammatory response in a subject in need thereof, wherein the method comprises or alternatively essentially consists of or additionally consists of administering to the subject an effective amount of an antibody or antigen-binding fragment thereof that binds to the head region of a DNABII polypeptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region and/or the IhfA5-mIhfB4NTHI chimeric head peptide). The DNABII polypeptide may be an IHF or HU peptide. These methods can be combined with diagnostic methods for detecting and/or monitoring the release or levels of cytokines in the tissues of the subject. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as described herein, which in one aspect specifically binds to the tail region or tail fragment of a DNABII polypeptide (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label. Non-limiting examples of cytokines include, for example, IL1beta, IL6, IL8, IL12p70, IL10, IL13, and IFN.

Additionally or alternatively, methods are provided for detecting a biofilm on a surface, comprising or alternatively consisting of or further consisting of contacting a surface (in one aspect susceptible to a biofilm or containing a biofilm) with an effective amount of one or more compounds selected from an antibody, an antigen-binding fragment, a polypeptide, or a CDR as described herein, wherein the antibody, fragment, polypeptide, or CDR thereof binds the tail or head region of a DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In one embodiment, the contacting is performed in vivo or in vitro. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or consisting essentially of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 4, 5 or 6, or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or consisting essentially of or additionally consisting of an amino acid sequence selected from the group of sequences of SEQ ID NO: 10, 11 or 12, or sequences equivalent to each of said sequences. In yet another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

Additionally or alternatively, a method for detecting a microbial infection that leads to the formation of a biofilm in a subject is provided. The method comprises or alternatively consists of or additionally consists of contacting an effective amount of one or more antibodies, fragments thereof, polypeptides or CDRs, as disclosed herein, with a biological sample suspected of containing a biofilm and isolated from the subject's body, and detecting binding of the antibody, fragment thereof, polypeptide or CDR to any biofilm in the sample. In one embodiment, the antibody, fragment thereof, polypeptide or CDR binds to the tail or head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, the contacting is carried out in vivo or in vitro. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of an immunoglobulin heavy chain (HC) comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of an immunoglobulin light chain (LC) comprising or essentially consisting of or additionally consisting of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or consisting essentially of or additionally consisting of amino acid sequences selected from the group of sequences of SEQ ID NO: 4, 5 or 6 or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or consisting essentially of or additionally consisting of amino acid sequences selected from the group of sequences of SEQ ID NO: 10, 11 or 12 or sequences equivalent to each of said sequences. In another aspect, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

Additionally or alternatively, a method for screening subjects having a biofilm is provided, comprising or alternatively consisting of or further consisting of contacting an effective amount of one or more antibodies, fragments thereof, polypeptides or CDRs, as disclosed herein, with a biological sample containing a biofilm and isolated from the subject's body, and detecting binding of the antibody, fragment thereof, polypeptide or CDR to any biofilm in the sample. In one embodiment, the antibody, fragment thereof, polypeptide or CDR binds to the tail or head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, the IhfA5-mIhfB4NTHI chimeric head peptide, the IHF or HU tail region, the IHFA or IHFB tail region and/or the IhfA3-IhfB2NTHI chimeric tail peptide). In another embodiment, a subject in whom binding is detected is selected for administration of an effective amount of one or more antibodies, fragments thereof, polypeptides or CDRs as disclosed herein, and/or an effective amount of one or more compounds selected from polynucleotides or vectors encoding an antibody, fragment thereof, polypeptide or CDR, wherein the antibody, fragment thereof, polypeptide or CDR binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region or the IhfA5-mIhfB4NTHI chimeric head peptide). In another embodiment, the contacting is carried out in vivo or in vitro. In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 26 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 27 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or consisting essentially of or additionally consisting of amino acid sequences selected from the group of sequences of SEQ ID NO: 4, 5 or 6 or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or consisting essentially of or additionally consisting of amino acid sequences selected from the group of sequences of SEQ ID NO: 10, 11 or 12 or sequences equivalent to each of said sequences. In another aspect, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

Furthermore, a method for producing an interfering nucleic acid is provided, comprising producing a nucleic acid consisting of about 10-20 nucleotides that specifically binds a specific binding partner to an antibody or fragment thereof, as disclosed in the present description, and, optionally, isolating the interfering nucleic acid obtained by this method.

When used in vitro, some of the disclosed methods are useful for screening or confirming agents (e.g., antibodies, fragments, and mimetics thereof) that have the same, similar, or opposite ability as the polypeptides, polynucleotides, antibodies, host cells, small molecules, and compositions disclosed herein. Alternatively, the methods can be used to determine which agent is best suited for treating a microbial infection or whether the treatment has been effective. For example, new agents or combination therapies can be screened by having two samples containing, for example, a DNABII polypeptide and microbial DNA, and a test agent. The DNABII polypeptide can be an IHF or HU peptide. The second sample contains the DNABII polypeptide and microbial DNA and a known active agent, for example, an agent described above, for example, an antibody or an antigen-binding fragment thereof, which serve as a positive control. In a further aspect, multiple samples are provided, and the agents are added to the system in increasing dilutions to determine the optimal dose likely to be effective in treating a subject in a clinical setting. As will be apparent to those skilled in the art, a negative control containing a DNABII polypeptide and microbial DNA can be provided. In yet another aspect, the DNABII polypeptide and microbial DNA are labeled with a detectable label, such as luminescent molecules that emit a signal upon close contact with each other. The samples are maintained under similar conditions for a time effective for the agent to inhibit, compete, or titrate the interaction between the DNABII polypeptide and microbial DNA, and the samples are then analyzed for signal emission from the luminescent molecules. If the sample generates a signal, the agent is ineffective in inhibiting binding.

In another embodiment, the in vitro method is used in a miniature well-slide system, wherein a microbial (such as a bacterial) isolate causing an infection can be isolated from a human/animal host and then cultured to ensure its growth as a biofilm in vitro. An agent (e.g., an antibody or antigen-binding fragment thereof) or a potential agent is added, alone or in combination with another agent, to the culture, with or without increasing dilutions of the potential agent or agent, such as an antibody or antigen-binding fragment thereof, to find the optimal dose that would likely be effective in treating the respective patient when delivered to a subject with an infection. As will be apparent to those skilled in the art, positive and negative controls can be performed simultaneously.

In a further aspect, the method is practically carried out on a high-throughput platform with an agent as described above, for example, an antibody or an antigen-binding fragment thereof, and/or a potential agent (alone or in combination with another agent) in a flow cell. The agent as described above, for example, an antibody or an antigen-binding fragment thereof, or a potential agent is added alone or in combination with another agent to the culture with or without increasing dilutions of the potential agent or agent as described above, for example, an antibody or an antigen-binding fragment thereof (or another antibody, small molecule, agent, etc.), to find the optimal dose that is likely to be effective in treating a patient when delivered to a subject with an existing infection. Biofilm isolates are sonicated to separate biofilm bacteria from a DNABII polypeptide, such as IHF, associated with microbial DNA. DNABII polypeptide-DNA complexes are isolated on the platform using an antibody against DNABII or IHF. Microbial DNA is then released, for example, by a salt wash, and used to identify the added biofilm bacteria. The released DNA is then identified, for example, by PCR sequencing. If DNA is not released, then the agent(s) have successfully bound or bound the microbial DNA. If DNA is detected in the sample, then the agent does not interfere with the binding of the DNABII polypeptide to the microbial DNA. As will be appreciated by those skilled in the art, positive and/or negative controls can be run simultaneously.

The above methods can also be used as a diagnostic test, since it is possible that a given bacterial species will respond better to the inversion of its biofilm by one agent than another; this rapid, high-throughput assay system could allow a person skilled in the art to analyze panels of possible anti-DNABII or IHF-like agents to identify the most effective ones from the group.

The advantage of these methods is that most hospital clinical microbiology laboratories are already equipped to perform such assays (i.e., determining MIC and MBC values) using bacteria grown in liquid culture (or planktonically). As is clear to those skilled in the art, bacteria typically do not grow planktonically when they cause disease. Instead, they grow as stable biofilms, and these biofilms are significantly more resistant to treatment with antibiotics, antibodies, or other therapeutics. This resistance is the reason why most MIC/MBC values cannot accurately predict in vivo efficacy. Therefore, by determining what "dose" of an agent can disrupt a bacterial biofilm in vitro (as described above), the preclinical assay proposed by the applicants could become a more reliable predictor of clinical efficacy, even for use in personalized medicine.

In addition to clinical settings, these methods can be used to identify the microbe causing the infection and/or confirm the effectiveness of treatments and agents in industrial settings. Thus, the agents can be used to treat, inhibit, or disrupt biofilms in industrial settings.

In another embodiment of the above methods, an antibiotic or antimicrobial agent known to inhibit the underlying infection is added sequentially or simultaneously to determine whether the infection can be suppressed. An interfering agent can also be added to the microbial DNA or DNABII polypeptide before adding the missing complex to assay for biofilm inhibition. In one aspect, DNase treatment is omitted from the method.

When used in vivo in non-human animals, such as chinchillas, the method provides preclinical screening to identify agents that can be used alone or in combination with other agents to disrupt biofilms.

In another embodiment, the present application provides a method for inhibiting, preventing, or disrupting a biofilm in a subject by administering to the subject an effective amount of an agent as described above, for example, an antibody or an antigen-binding fragment thereof, thereby inhibiting, preventing, or disrupting a microbial biofilm. The methods comprise or consist essentially of or consist of administering to the subject an antibody or fragment thereof as disclosed herein that binds to the head region of a DNABII peptide (including, but not limited to: an IHF or HU head region, an IHFA or IHFB head region, and/or an IhfA5-mIhfB4NTHI chimeric head peptide). The DNABII peptide may be an IHF or HU peptide. The antibody or antigen-binding fragment thereof may be labeled with a detectable label. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or disruption. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as disclosed herein, which in one aspect specifically binds to the tail region or fragment of a DNABII polypeptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, or the tail chimeric peptide IhfA3-IhfB2NTHI). In another embodiment, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

In one embodiment, the agent is one or more antibodies and/or antigen-binding fragments, which are the same or different from each other. In some embodiments, such antibodies or antigen-binding fragments are administered alone or in combination with each other, or with an agent other than the antibody, or with an additional pharmaceutically effective agent, alone or in combination with a pharmaceutically acceptable carrier. Non-limiting examples of such subjects include mammals, such as domestic animals, and human patients.

The present application also provides a method for inducing an immune response or generating passive immunity in a subject in need thereof, comprising or alternatively essentially consisting of or additionally consisting of administering to the subject an effective amount of one or more antibodies or antigen-binding fragments thereof, as disclosed herein, that bind to a head region of a DNABII peptide, such as an IHF or HU peptide. Such a head region of a DNABII peptide includes, but is not limited to: an IHF or HU head region, an IHFA or IHFB head region, or an IhfA5-mIhfB4NTHI chimeric head peptide. In one embodiment, a method for generating passive immunity in a subject is provided, comprising or alternatively essentially consisting of or further consisting of administering to the subject an effective amount of one or more antibodies, fragments thereof, polypeptides, or CDRs as disclosed herein, and/or an effective amount of one or more compounds selected from polynucleotides or vectors encoding the antibody, fragment thereof, polypeptide, or CDR, wherein the antibody, fragment, polypeptide, or CDR binds to the head region of the DNABII peptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, or the IhfA5-mIhfB4NTHI chimeric head peptide). In one embodiment, an antibody or fragment thereof is provided that comprises or essentially consists of or additionally consists of a sequence of a variable domain of a heavy chain (HC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 24 or a sequence equivalent thereto, and/or a sequence of a variable domain of a light chain (LC) of an immunoglobulin comprising or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 25 or a sequence equivalent thereto. In another embodiment, the antibody or fragment thereof comprises or consists essentially of or additionally consists of an immunoglobulin heavy chain (HC) variable domain sequence comprising or consisting essentially of or additionally consisting of amino acid sequences selected from the group of sequences of SEQ ID NO: 1, 2 or 3 or sequences equivalent to each of said sequences, and/or an immunoglobulin light chain (LC) variable domain sequence comprising or consisting essentially of or additionally consisting of amino acid sequences selected from the group of sequences of SEQ ID NO: 7, 8 or 9 or sequences equivalent to each of said sequences. In another aspect, the antibody or antigen-binding fragment thereof can be labeled with a detectable label. These methods can be combined with diagnostic methods for detecting and/or monitoring biofilm formation and/or destruction. In one embodiment, the diagnostic methods comprise using an antibody or antigen-binding fragment as described herein, which in one aspect specifically binds to the tail region or tail fragment of a DNABII polypeptide (including, but not limited to: the tail region of IHF or HU, the tail region of IHFA or IHFB, or the tail chimeric peptide IhfA3-IhfB2NTHI). In another aspect, the antibody or antigen-binding fragment thereof is labeled with a detectable label.

In one embodiment, the agent is one or more antibodies and/or antigen-binding fragments, which are the same or different from each other. In some embodiments, such antibodies or antigen-binding fragments are administered alone or in combination with each other, or with an agent other than the antibody, or with an additional pharmaceutically effective agent, alone or in combination with a pharmaceutically acceptable carrier.

In a further embodiment, the methods further comprise or, alternatively, consist essentially of or further consist of administering to the subject an effective amount of one or more antimicrobial agents, antigenic peptides, or adjuvants.

A non-limiting example of an antimicrobial agent is another component of the vaccine, such as a surface antigen, e.g., OMP P5 protein, rsPilA, OMP 26, OMP P2, or pilin type IV (see Jurcisek and Bakaletz (2007) J. Bacteriology 189(10):3868-3875; Murphy, T.F. et al. (2009) The Pediatric Infectious Disease Journal 28:S121-S126).

The agents and compositions disclosed herein can be administered simultaneously or sequentially with other antimicrobial agents and/or surface antigens. In one specific embodiment, administration is local to the site of infection, such as by direct injection or inhalation. Other non-limiting examples of administration include one or more routes including transdermal, urethral, sublingual, rectal, vaginal, ocular, subcutaneous, intramuscular, intraperitoneal, intranasal, inhalation, or oral.

Microbial infections and diseases that can be treated by the methods disclosed in this application include infection with a gram-positive or gram-negative biofilm-forming organism, such as Streptococcus agalactiae, Neisseria meningitidis, Treponemes denticola pallidum, Burkholderia cepacia, or Burkholderia pseudomallei. In one embodiment, the microbial infection is one or more of Haemophilus influenzae (non-encapsulated types), Moraxella catarrhalis, Streptococcus pneumoniae, Streptococcus pyogenes, Pseudomonas aeruginosa, or Mycobacterium tuberculosis. These microbial infections may be present in the upper, middle, and lower respiratory tract (otitis, sinusitis, bronchitis, as well as exacerbations of chronic obstructive pulmonary disease (COPD), chronic cough, complications and/or underlying cause of cystic fibrosis (CF), and community acquired pneumonia (CAP). Thus, using the methods disclosed herein in vivo, it is also possible to prevent or treat these diseases and complications of these infections.

Infections can also occur in the oral cavity (caries, periodontitis) and are caused by Streptococcus mutans, Porphyromonas gingivalis, and Aggregatibacter actinomvctemcomitans. Infections can also be localized on the skin (abscesses, staph infections, impetigo, secondary burn infections, Lyme disease) and are caused by Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Borrelia burdorferi. Urinary tract infections (UTIs) are also treatable and are usually caused by E. coli. Gastrointestinal tract (GI) infections (diarrhea, cholera, gallstones, gastric ulcers) are commonly caused by Salmonella enterica serovar, Vibrio cholerae, and Helicobacter pylori. Genital tract infections are commonly caused by Neisseria gonorrhoeae. Bladder infections or those associated with implanted devices can be caused by Enterococcus faecalis. Infections associated with implanted prosthetic devices, such as hip or knee joint replacements, or dental implants, or medical devices such as pumps, catheters, stents, or monitoring systems, commonly caused by various bacteria, can be treated using the methods described herein. These devices can be coated or conjugated with an agent as disclosed herein. Thus, by applying the methods disclosed herein in vivo, the aforementioned diseases and complications of these infections can also be prevented or treated.

Infections caused by Streptococcus agalactiae can also be treated with the methods described herein and are the leading cause of bacterial sepsis in neonates. Infections caused by Neisseria meningitidis, which can cause meningitis, can also be treated.

Thus, the routes of administration utilized in the methods disclosed herein include intranasal, intramuscular, urethral, intratracheal, subcutaneous, intradermal, transdermal, topical, intravenous, rectal, nasal, oral, inhalation, and other enteral and parenteral routes of administration. The routes of administration may be combined or adjusted, if desired, depending on the agent and/or the desired effect. The active agent may be administered as a single dose or in multiple doses. Embodiments of these methods and routes suitable for delivery include systemic or local routes. Typically, routes of administration suitable for the methods described herein include, but are not limited to, direct injection, enteral, parenteral, or inhalation routes.

Parenteral routes of administration other than inhalation include, but are not limited to, topical, transdermal, subcutaneous, intramuscular, intraorbital, intracapsular, intraspinal, intrasternal, and intravenous routes—i.e., any route of administration other than via the alimentary canal. Parenteral administration can be used to achieve systemic or local delivery of the inhibitory agent. When systemic delivery is desired, administration typically involves invasive or local administration via systemic abrasion or transmucosal administration of pharmaceutical formulations.

The agents disclosed herein can also be delivered to a subject via enteral administration. Enteral routes of administration include, but are not limited to, oral and rectal (e.g., via a suppository).

Methods of administering the active substance through the skin or mucous membrane include, but are not limited to, topical application of a suitable pharmaceutical formulation, transdermal delivery, transdermal delivery, injection, and epidermal administration. Suitable methods for transdermal delivery include absorption promoters or iontophoresis. Iontophoretic delivery can be accomplished using commercially available "patches," which continuously deliver the appropriate product via electrical impulses through intact skin for several days or more.

In various embodiments of the methods disclosed herein, the interfering agent will be administered by inhalation, injection, or orally on a continuous daily basis, at least once per day (QD), and in various embodiments, two (BID), three (TID), or even four times per day. Typically, a therapeutically effective daily dose will be at least about 1 mg, or at least about 10 mg, or at least about 100 mg, or from about 200 to about 500 mg, and sometimes, depending on the compound, up to from about 1 g to about 2.5 g.

The dosage may be carried out in accordance with the methods disclosed herein using capsules, tablets, oral suspension, suspension for intramuscular injection, suspension for intravenous infusion, gel or cream for topical application, or suspension for intra-articular injection.

The dosage, toxicity, and therapeutic efficacy of the compositions described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD ₅₀ (the dose lethal to 50% of the population) and ED ₅₀ (the dose therapeutically effective to 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD ₅₀ /ED ₅₀ . In some embodiments, the compositions exhibit high therapeutic indices. Although compounds that exhibit toxic side effects can be used, care should be taken to develop a delivery system that targets such compounds to the site of diseased tissue in order to minimize potential damage to uninfected cells and thus reduce side effects.

Data obtained from cell culture assays and animal studies can be used to determine a dosage range for use in humans. The dosage of such compounds is (in some embodiments) within a range of circulating concentrations that includes the ED ₅₀ with little or no toxicity. The dosage may vary within this range depending on the dosage form and route of administration used. For any compound used in the methods, the therapeutically effective dose can be initially estimated based on cell culture assays. The dose can be determined in animal models to achieve a range of circulating plasma concentrations that includes the IC ₅₀ (i.e., the concentration of the test compound at which half-maximal inhibition of symptoms is achieved), as determined in cell culture. Such information can be used to more accurately determine useful doses for humans. Plasma levels can be measured, for example, by high-performance liquid chromatography.

In some embodiments, an effective amount of the composition sufficient to achieve a therapeutic or prophylactic effect ranges from about 0.000001 mg per kilogram of body weight per administration to about 10,000 mg per kilogram of body weight per administration. Suitable dosage ranges are from about 0.0001 mg per kilogram of body weight per administration to about 100 mg per kilogram of body weight per administration. Administration can be provided as an initial dose followed by one or more "booster" doses. Booster doses can be administered one day, two days, three days, one week, two weeks, three weeks, one, two, three, six, or twelve months after the initial dose. In some embodiments, the booster dose is administered after assessing the subject's response to the previous doses.

A skilled artisan will understand that certain factors may influence the dosage and timing necessary to effectively treat a subject, including, but not limited to, the severity of the disease or disorder, prior treatment, the general health and/or age of the subject, and the presence of other medical conditions. Furthermore, administration of a therapeutically effective amount of the therapeutic compositions described herein to a subject may involve a single administration or a series of administrations.

Polypeptides

Also provided in the present application are isolated polypeptides comprising heavy and light chains of antibodies, their antigen-binding fragments, CDRs and equivalents of each of said compounds with the addition of up to 25, or alternatively 20, or alternatively 15, or alternatively up to 10, or alternatively up to 5 random amino acids at either the amino or carboxyl terminus (or at both termini).

In one aspect, the present application provides isolated polypeptides comprising or consisting essentially of, or additionally consisting of, an amino acid sequence from the group of SEQ ID NOs: 1-14 or 24-27, or an equivalent of each. The polypeptides may further comprise a detectable marker or a purification marker.

The present invention also provides isolated or recombinant polypeptides comprising or alternatively consisting essentially of or additionally consisting of two or more, or three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more of all fourteen isolated polypeptides or fragments or equivalents of each of them.

In any of the above embodiments, a peptide linker may be added to the N-terminus or C-terminus of the polypeptide. "Linker" or "peptide linker" refers to a peptide sequence linked to either the N-terminus or C-terminus of a polypeptide sequence. In one aspect, the linker is from about 1 to about 20 amino acid residues in length, or alternatively from 2 to about 10, about 3 to about 5 amino acid residues. An example of a peptide linker is Gly-Pro-Ser-Leu-Lys-Leu (SEQ ID NO: 33). Other examples include Gly-Gly-Gly; Gly-Pro-Ser-Leu (SEQ ID NO: 34); Gly-Pro-Ser; Pro-Ser-Leu-Lys (SEQ ID NO: 35); Gly-Pro-Ser-Leu-Lys (SEQ ID NO: 36) and Ser-Leu-Lys-Leu (SEQ ID NO: 37).

The isolated polypeptides described herein include recombinantly produced polypeptides and proteins from prokaryotic and eukaryotic host cells, as well as muteins, analogs, and fragments thereof, examples of which are described above. In some embodiments, such peptides also include antibodies and anti-idiotypic antibodies, as described herein. Such polypeptides can be isolated or produced using methods known in the art and briefly described herein.

It is understood that functional equivalents or variants of a wild-type polypeptide or protein are also within the scope of the present invention, for example, having conservative amino acid substitutions.

In a further aspect, the polypeptides are conjugated or linked to a detectable label or agent for increasing the half-life of the polypeptide, for example, by PEGylation, PEGmimetation, polysialylation, HESylation, or glycosylation. Suitable labels are known in the art and are described herein.

In another aspect, the polypeptides, with or without a detectable label, may be present in or expressed on the surface of a prokaryotic or eukaryotic host cell, such as a dendritic cell.

Proteins and polypeptides can be produced by a variety of methods known to those skilled in the art, including purification, chemical synthesis, and recombinant techniques. Polypeptides can be isolated from preparations such as host cell systems by methods such as immunoprecipitation with antibody and standard techniques such as gel filtration, ion exchange, reverse phase, and affinity chromatography. Such techniques are described, for example, in Deutscher et al. (1999) Guide To Protein Purification: Methods In Enzymology (Vol. 182, Academic Press). Accordingly, the present invention also provides methods for producing said polypeptides, as well as the products obtained by these methods.

Polypeptides can also be produced by chemical synthesis using a commercially available automated peptide synthesizer, such as the Perkin/Elmer/Applied Biosystems, Inc., model 430A or 431A, Foster City, California, USA. The synthesized polypeptide can be precipitated and further purified, for example, by high-performance liquid chromatography (HPLC). Accordingly, the present invention also provides a method for chemically synthesizing the proteins disclosed herein by providing protein sequences and reagents, such as amino acids and enzymes, and linking the amino acids together in the proper orientation and linear sequence.

Alternatively, proteins and polypeptides can be produced by well-known recombinant methods, as described, for example, in Sambrook et al. (1989), supra, using the host cells and vector systems disclosed herein.

This application also provides the polypeptides disclosed herein conjugated to a detectable agent for use in diagnostic methods. For example, detectably labeled polypeptides can be coupled to a column and used to detect and purify antibodies. They are also useful as immunogens for producing antibodies. The polypeptides described herein are useful in an in vitro assay system for screening agents or drugs that modulate cellular processes. In another aspect, antibodies specific for the tail regions of DNABII polypeptides (including, but not limited to: the IHF or HU tail region, the IHFA or IHFB tail region, and/or the IhfA3-IhfB2NTHI tail chimeric peptide) are particularly useful in diagnostic assays for detecting biofilms and can be used alone or in combination with one or more antibodies as described herein. In one aspect, antibodies specific to the tail regions are used as companion diagnostics for an antibody specific to the head region of a DNABII polypeptide (including, but not limited to: the IHF or HU head region, the IHFA or IHFB head region, and/or the IhfA5-mIhfB4NTHI head chimeric peptide). The DNABII polypeptide may be an IFH or HU polypeptide.

Those skilled in the art are well aware that modifications can be made to the peptides described herein to impart altered properties. As used herein, the term "amino acid" refers to either natural and/or unnatural or synthetic amino acids, including glycine and both D or L optical isomers, as well as amino acid analogs and peptidomimetics. A peptide of three or more amino acids is typically called an oligopeptide if the peptide chain is short. If the peptide chain is long, the peptide is typically called a polypeptide or protein.

The peptides described in this application can be modified by the inclusion of unnatural amino acids. Thus, the peptides can contain D-amino acids, a combination of L-amino acids, and various "designer" amino acids (e.g., beta-methyl-amino acids, C-alpha-methyl-amino acids, and N-alpha-methyl-amino acids, etc.) to impart specific properties to the peptides. Furthermore, by incorporating specific amino acids at specific stages of coupling, peptides with alpha helices, beta turns, beta sheets, gamma turns, and cyclic peptides can be created. It is generally believed that an alpha-helical secondary structure or a randomized secondary structure can be particularly useful.

The polypeptides described in this application can also be combined with various solid-phase carriers, such as an implant, stent, paste, gel, dental implant, or medical implant, or liquid-phase carriers, such as beads, sterile or aqueous solutions, pharmaceutically acceptable carriers, pharmaceutically acceptable polymers, liposomes, micelles, suspensions, and emulsions. Examples of non-aqueous solvents include propylethylene glycol, polyethylene glycol, and vegetable oils. When used to produce antibodies or induce an immune response in vivo, the carriers can also include an adjuvant that is useful for non-specifically enhancing a specific immune response. A skilled artisan can readily determine whether an adjuvant is required and select one. However, for illustrative purposes only, suitable adjuvants include, but are not limited to, complete and incomplete Freund's adjuvants, mineral salts, and polynucleotides. Other suitable adjuvants include monophosphoryl lipid A (MPL), mutant derivatives of E. coli heat-labile enterotoxin, mutant derivatives of cholera toxin, CPG oligonucleotides, and squalene-derived adjuvants.

The present invention also provides a pharmaceutical composition comprising, or alternatively essentially consisting of, any polypeptide, analog, mutein, or fragment disclosed herein, alone or in combination with each other or with other agents, such as an antibiotic and an acceptable carrier or solid support. These compositions are useful for the various diagnostic and therapeutic methods described herein.

Polynucleotides

The present invention also provides isolated or recombinant polynucleotides encoding one or more of the above-identified antibodies, fragments thereof, CDRs, isolated or recombinant polypeptides, and their corresponding complementary chains. Vectors containing isolated or recombinant polynucleotides are also examples known in the art and are briefly described herein. In one aspect, when more than one isolated or recombinant polynucleotide is to be expressed as a unit, the isolated or recombinant polynucleotides can be contained in a polycistronic vector. The polynucleotides can be DNA, RNA, mRNA, or interfering RNA, such as small interfering RNA, microRNA, or double-stranded RNA.

In another embodiment, the present invention provides an interfering agent that is a polynucleotide that interferes with the binding of DNA to a polypeptide or protein in a microbial biofilm, or a four-way junction polynucleotide resembling a Holliday junction, a three-way junction polynucleotide resembling a replication fork, a polynucleotide having inherent flexibility, or a bent polynucleotide that can inhibit the binding of a DNABII (HU or IHF) polynucleotide to microbial DNA and treat, prevent, or inhibit the formation of biofilms and associated infections and disorders. One skilled in the art can prepare such polynucleotides using the information provided herein and the knowledge of those skilled in the art. See Goodman and Kay (1999) J. Biological Chem. 274(52):37004-37011 and Kamashev and Rouviere-Yaniv (2000) EMBO J. 19(23):6527-6535.

The application further provides an isolated or recombinant polynucleotide operably linked to an RNA transcription promoter, as well as other regulatory sequences for the replication and/or transient or stable expression of DNA or RNA. As used herein, the term "operably linked" means positioned such that the promoter will direct the transcription of RNA from a DNA molecule. Examples of such promoters are SP6, T4, and T7. In some embodiments, cell-specific promoters are used for cell-specific expression of the inserted polynucleotide. Vectors that contain a promoter or promoter/enhancer with stop codons and selectable marker sequences, as well as a cloning site into which an inserted DNA fragment can be operably linked to this promoter, are known in the art and are commercially available. For general cloning methodology and strategy, see Gene Expression Technology (Goeddel ed., Academic Press, Inc. (1991)) and references therein, and Vectors: Essential Data Series (Gacesa and Ramji, eds., John Wiley & Sons, N.Y. (1994)), which contains maps, functional properties, commercial suppliers, and a link to GenEMBL accession numbers for various suitable vectors.

In one embodiment, polynucleotides derived from the polynucleotides disclosed herein encode polypeptides or proteins having diagnostic and therapeutic properties as described herein, and also serve as probes for identifying protein transcripts, which may or may not be present. These nucleic acid fragments can be obtained, for example, by restriction enzyme digestion of large polynucleotides and then labeled with a detectable marker. Alternatively, randomized fragments can be generated using nick translation of the molecule. For methodology for obtaining and labeling such fragments, see Sambrook et al. (1989) supra.

Expression vectors containing these nucleic acids are useful for constructing host vector systems for the production of proteins and polypeptides. These expression vectors are intended to replicate in host organisms either as episomes or as an integral part of chromosomal DNA. Non-limiting examples of suitable expression vectors include plasmids, yeast vectors, viral vectors, and liposomes. Adenoviral vectors are particularly useful for introducing genes into tissues in vivo due to their high expression levels and efficient cell transformation both in vitro and in vivo. When the nucleic acid is inserted into a suitable host cell, such as a prokaryotic or eukaryotic cell, and the host cell replicates, the protein can be produced recombinantly. The selection of suitable host cells will depend on the vector and such vectors may include mammalian cells, animal cells, human cells, monkey cells, insect cells, yeast cells, and bacterial cells constructed using known methods. See Sambrook et al. (1989) supra. In addition to using a viral vector to introduce exogenous nucleic acid into cells, the nucleic acid can be delivered into the host cell by methods known in the art, such as transformation of bacterial cells, transfection using calcium phosphate precipitation for mammalian cells or DEAE-dextran, by electroporation, or microinjection. For methodology, see Sambrook et al. (1989) supra. Thus, the present invention also relates to a host cell, such as a mammalian cell, an animal cell (rat or mouse), a human cell, or a prokaryotic cell such as a bacterial cell, containing a polynucleotide encoding a protein or polypeptide or an antibody.

A polynucleotide may contain modified nucleotides, such as methylated nucleotides and nucleotide analogs. If modified nucleotides are used, modifications to the nucleotide structure may be provided before or after assembly of the polynucleotide. The nucleotide sequence may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, for example, by conjugation with a labeling component. The term also refers to both double-stranded and single-stranded molecules. Unless otherwise stated or required, any embodiment disclosed herein that is a polynucleotide encompasses both the double-stranded form and each of the two complementary single-stranded forms known or suspected to constitute the double-stranded form.

When vectors are used in the method disclosed herein for in vivo or ex vivo gene therapy, a pharmaceutically acceptable vector, such as a replication-incompetent retroviral or adenoviral vector, is illustrative (but not limiting) and may be intended for a specific use. Pharmaceutically acceptable vectors containing the nucleic acids disclosed herein may be further modified for transient or stable expression of the inserted polynucleotide. The term "pharmaceutically acceptable vector" as used herein includes, but is not limited to, a vector or delivery vehicle having the ability to selectively target and introduce nucleic acid into dividing cells. An example of such a vector is a "replication-incompetent" vector, defined by its inability to produce viral proteins, which prevents the vector from spreading in an infected host cell. An example of a replication-incompetent retroviral vector is LNL6 (Miller et al. (1989) BioTechniques 7:980–990). Methodologies for using replication-incompetent retroviruses for retroviral-mediated transfer of marker genes have been developed (Bordignon (1989) PNAS USA 86:8912–8952; Culver (1991) PNAS USA 88:3155; and Rill (1991) Blood 79(10):2694–2700).

The present application also relates to genetically modified cells that contain and/or express the polynucleotides disclosed herein. Genetically modified cells can be produced by inserting upstream regulatory sequences, such as promoters or gene activators (see U.S. Patent No. 5,733,761). In one embodiment, the modified cells are eukaryotic cells or prokaryotic cells.

Polynucleotides can be conjugated to a detectable marker, such as an enzymatic label or a radioisotope, for detecting nucleic acid and/or gene expression in a cell. A wide range of suitable detectable markers are known in the art, including fluorescent, radioactive, enzymatic, or other ligands, such as avidin/biotin, that are capable of producing a detectable signal. In one aspect, it will likely be desirable to use a fluorescent label or an enzymatic label, such as urease, alkaline phosphatase, or peroxidase, instead of radioactive or other environmentally undesirable reagents. In the case of enzymatic labels, calorimetric indicator substrates can be used to provide means visible to the human eye or spectrophotometrically to identify specific hybridization with samples containing complementary nucleic acids. Thus, the present invention further provides a method for detecting a single-stranded polynucleotide or a polynucleotide complementary thereto by contacting a target single-stranded polynucleotide with a labeled single-stranded polynucleotide (probe), which is part of a polynucleotide disclosed herein, under conditions allowing hybridization (optionally under moderately stringent hybridization conditions) of complementary single-stranded polynucleotides or optionally under very stringent hybridization conditions. Hybridized polynucleotide pairs are separated from unhybridized single-stranded polynucleotides. Hybridized polynucleotide pairs are detected using methods known to those skilled in the art and described, for example, in Sambrook et al. (1989) supra.

The polynucleotide according to the present invention can be produced using chemical synthesis, recombinant cloning methods, PCR, or any combination thereof. Methods for chemically synthesizing polynucleotides are known in the art and need not be described in detail herein. One skilled in the art can use the sequence data provided herein to produce the desired polynucleotide using a DNA synthesizer or by ordering from a commercial service.

The polynucleotides disclosed herein can be isolated or replicated using PCR. PCR technology is the subject of invention in U.S. Patents 4,683,195; 4,800,159; 4,754,065; and 4,683,202 and is described in "PCR: The Polymerase Chain Reaction" (Mullis et al. eds., Birkhauser Press, Boston (199.4)) or in MacPherson et al. (1991) and (1995) (see above), and references cited therein. Alternatively, one skilled in the art can use the sequences disclosed herein and a commercial DNA synthesizer to replicate DNA. Accordingly, the present invention also provides a method for producing the polynucleotides disclosed herein by providing a linear polynucleotide sequence, nucleotides, appropriate primer molecules, chemicals such as enzymes, and instructions for replicating them, and chemically replicating or linking the nucleotides in the proper orientation to produce the polynucleotides. In a particular embodiment, these polynucleotides are further isolated. Furthermore, one skilled in the art can insert the polynucleotide into a suitable replication vector and insert the vector into a suitable host cell (prokaryotic or eukaryotic) for replication and amplification. The DNA thus amplified can be isolated from the cell by methods known to those skilled in the art. The present description further provides a method for producing polynucleotides, as well as polynucleotides obtained by this method.

RNA can be produced by first inserting a DNA polynucleotide into a suitable host cell. The DNA can be delivered by any suitable method, such as using a suitable gene delivery vehicle (e.g., liposomes, plasmids, or vectors) or by electroporation. Once the cell replicates and the DNA is transcribed into RNA, the RNA can then be isolated using methods known to those skilled in the art, such as those described in Sambrook et al. (1989) supra. For example, mRNA can be isolated using various lytic enzymes or chemical solutions according to the procedures described in Sambrook et al. (1989), supra, or extracted with nucleic acid binding resins according to the accompanying instructions provided by the manufacturers.

Polynucleotides having a sequence complementary to or homologous to the sequence of the polynucleotide described in this application can be used as hybridization probes or as equivalents to the specific polynucleotides disclosed herein. Since the complete coding sequence of the transcript is known, any portion of this sequence or homologous sequences can be used in the methods disclosed herein.

It is known in the art that a "perfectly matched" probe is not required for specific hybridization. Minor changes in the probe sequence, achieved by substitution, deletion, or insertion of a small number of bases, do not affect the specificity of hybridization. Generally, up to 20% mismatch in base pairs is acceptable (with optimal alignment). In some embodiments, a probe suitable for detecting the aforementioned mRNA is at least about 80% identical to the homologous region. In some embodiments, the probe is 85% identical to the corresponding gene sequence after alignment of the homologous region; in some embodiments, the probe exhibits 90% identity.

These probes can be used in radioassays (e.g., Southern and Northern blotting) for the detection, prognosis, diagnosis, or monitoring of various cells or tissues containing these cells. The probes can also be attached to a solid support or matrix, such as a chip, for use in high-throughput screening assays to detect the expression of a gene corresponding to the polynucleotide disclosed herein. Accordingly, the present invention also provides a probe comprising or corresponding to the polynucleotide disclosed herein, or its equivalent, or its complement, or a fragment thereof, attached to a solid support for use in high-throughput screening.

The overall fragment size, as well as the size of the complementary regions, will depend on the intended use or application of the specific nucleic acid segment. Smaller fragments are typically used in hybridization methods, where the complementary region length can vary, for example, from at least 5 to 10 to approximately 100 nucleotides, or even the full length, depending on the complementary sequences to be detected.

Nucleotide probes having complementary sequences of regions longer than 5-10 nucleotides are generally well suited for increasing the stability and selectivity of the hybrid molecule and, thus, improving the specificity of the resulting specific hybrid molecules. In some embodiments, polynucleotides can be designed having gene complementary regions of 10 or more, or more than 50 nucleotides, or even longer, if necessary. Such fragments can be readily obtained, for example, by direct synthesis of the fragment by chemical means using nucleic acid reproduction technology, such as the two-prime oligonucleotide PCR technology, as described in U.S. Patent No. 4,603,102, or by introducing selected sequences into recombinant vectors for recombinant production. In one aspect, the probe length is about 50-75 or more, alternatively 50-100 nucleotides.

The polynucleotides of the present invention can serve as primers for detecting genes or gene transcripts expressed in the cells described herein. In this context, amplification refers to any method using a primer-dependent polymerase capable of replicating the target sequence with acceptable fidelity. Amplification can be accomplished using natural or recombinant DNA polymerases, such as T7 DNA polymerase, the Klenow fragment of E. coli DNA polymerase, and reverse transcriptase. For illustrative purposes only, the primer is the same length as the probes.

One method for amplifying polynucleotides is PCR, and PCR amplification kits are commercially available. After amplification, the resulting DNA fragments can be detected by any suitable method known in the art, such as agarose gel electrophoresis followed by visualization with ethidium bromide staining and ultraviolet illumination.

Methods for introducing an effective amount of a gene delivery vector or carrier into a cell have been developed and are known to those skilled in the art and are described in this application. Methods for detecting gene expression in a cell are known in the art and include methods such as DNA microarray hybridization, in situ hybridization, PCR, RNase protection assays, and Northern blotting. Such methods are useful for detecting and quantifying gene expression in a cell. Alternatively, expression of the encoded polypeptide can be detected by various methods. In particular, it is useful to generate polyclonal or monoclonal antibodies that specifically react with the target polypeptide. Such antibodies are useful for visualizing cells that express the polypeptide using methods such as immunohistology, ELISA, and Western blotting. These methods can be used to determine the expression level of the expressed polynucleotide.

Methods of obtaining

Also provided are methods for producing antibodies, fragments, CDRs, or polypeptides comprising or alternatively consisting of or additionally consisting of culturing a host cell comprising a polynucleotide encoding an antibody, antigen-binding fragment, polypeptide, or CDR under polynucleotide expression conditions and optionally isolating the antibody, fragment, CDR, and/or polypeptide from the cell and/or culture. Additionally provided is a host cell comprising a polynucleotide encoding an antibody, antigen-binding fragment, polypeptide, or CDR under polynucleotide expression conditions. In one embodiment, the host cell is a eukaryotic cell or a prokaryotic cell. In another embodiment, the host cell is a mammalian cell.

Compositions

Compositions are further provided. The compositions comprise a carrier and one or more components selected from an isolated polypeptide described herein, an isolated polynucleotide described herein, a vector described herein, an isolated host cell described herein, a small molecule, or an antibody and/or antigen-binding fragment described herein. The carriers may be one or more solid carriers or a pharmaceutically acceptable carrier. The compositions may further comprise an adjuvant or other components suitable for administration as vaccines. In one embodiment, the compositions comprise one or more pharmaceutically acceptable excipients, diluents, carriers, and/or adjuvants. In addition, the compositions of the present invention include one or more components selected from an isolated polypeptide described herein, an isolated polynucleotide described herein, a vector described herein, a small molecule, an isolated host cell described herein, or an antibody according to the present invention, and also include one or more pharmaceutically acceptable substances.

In the case of oral preparations, any one or more components selected from the isolated or recombinant polypeptide described herein, the isolated or recombinant polynucleotide described herein, the vector described herein, the isolated host cell described herein, the small molecule, or the antibody or fragment thereof described herein can be used alone or in the pharmaceutical compositions disclosed herein comprising or consisting essentially of the compound in combination with appropriate additives for the manufacture of tablets, powders, granules or capsules, for example with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrating agents such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants such as talc or magnesium stearate; and, if desired, with diluents, buffering agents, wetting agents, preservatives and flavorings. Pharmaceutically compatible binders and/or adjuvant materials may be included as part of the composition. Tablets, pills, capsules, lozenges and the like may contain any of the following ingredients or compounds of a similar nature: a binder such as microcrystalline cellulose, tragacanth, or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetener such as sucrose or saccharin; or a flavoring such as peppermint, methyl salicylate, or orange flavoring.

Pharmaceutical formulations and unit dosage forms suitable for oral administration are particularly useful in the treatment of chronic conditions, infections, and self-administered therapies. In one embodiment, the formulation is specific for pediatric administration.

The present invention provides pharmaceutical formulations in which one or more components of an isolated polypeptide disclosed herein, an isolated polynucleotide disclosed herein, a vector disclosed herein, an isolated host cell disclosed herein, or an antibody disclosed herein can be included in the injection formulations according to the invention by dissolving, suspending or emulsifying them in an aqueous or non-aqueous solvent, such as vegetable or other similar oils, synthetic glycerides of aliphatic acids, esters of higher aliphatic acids or propylene glycol, and, if necessary, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives or other antimicrobial agents. A non-limiting example of such an agent is an antimicrobial agent, such as other vaccine components such as surface antigens, e.g., OMP P5 protein, OMP 26, OMP P2, or type IV pilin (see Jurcisek and Bakaletz (2007) J. of Bacteriology 189(10):3868-3875 and Murphy, TF, Bakaletz, LO, and Smeesters, PR (2009) The Pediatric Infectious Disease Journal, 28:S121-S126) and antibacterial agents. For intravenous administration, suitable carriers include physiological bacteriostatic water, Cremophor EL™ (BASF, Parsippany, NJ), or phosphate-buffered saline (PBS). In all cases, the composition for parenteral administration should be sterile and fluid to the extent that it can be easily administered with a syringe.

The aerosol formulations described herein can be administered by inhalation and can be propellant-based or propellant-free. For example, embodiments of the pharmaceutical formulations disclosed herein include a compound disclosed herein combined with suitable propellants under pressure, such as dichlorodifluoromethane, propane, nitrogen, and the like. For administration by inhalation, the compounds can be delivered as an aerosol spray from a pressurized container or dispenser that contains a suitable propellant, such as carbon dioxide, or an atomizer. A non-limiting example of a non-propellant is a pump spray that is ejected from a closed container by mechanical force (i.e., by pressing a plunger with a finger or by squeezing the container, such as by applying a compressive force to the wall of the container or an elastic force created by the wall itself, such as an elastic bladder).

The suppositories disclosed herein can be prepared by mixing the compound disclosed herein with any of a variety of bases, such as emulsifying bases or water-soluble bases. Embodiments of such a pharmaceutical formulation of the compound disclosed herein can be administered rectally via a suppository. The suppository may contain excipients such as cocoa butter, carbowax, and polyethylene glycols, which melt at body temperature but solidify at room temperature.

Unit dosage forms for oral or rectal administration, such as syrups, elixirs, and suspensions, may be provided, wherein each unit form, for example, a teaspoon, tablespoon, tablet, or suppository, contains a predetermined amount of a composition comprising one or more compounds disclosed herein. Similarly, unit dosage forms for injection or intravenous administration may contain a compound disclosed herein as a solution in sterile water, saline, or another pharmaceutically acceptable carrier.

Embodiments of the pharmaceutical formulations disclosed herein include those in which one or more of an isolated polypeptide disclosed herein, an isolated polynucleotide disclosed herein, a vector disclosed herein, a small molecule for use in the invention, an isolated host cell disclosed herein, or an antibody or fragment thereof as disclosed herein are included in an injectable composition. The injectable pharmaceutical formulations disclosed herein are prepared as liquid solutions or suspensions, or as solid forms suitable for solution or suspension in liquid vehicles prior to injection. The formulation may also be emulsified or the active ingredient may be encapsulated in liposomal vehicles, in accordance with other embodiments of the pharmaceutical formulations disclosed herein.

In one embodiment, one or more of the isolated polypeptide disclosed herein, the isolated polynucleotide disclosed herein, the vector disclosed herein, the isolated host cell disclosed herein, or the antibody disclosed herein are formulated for delivery using a continuous delivery system. The term "continuous delivery system" is used herein interchangeably with "controlled delivery system" and encompasses continuous (e.g., controlled) delivery devices (e.g., pumps) in combination with catheters, injection devices, etc., a wide variety of which are known in the art.

Mechanical or electromechanical infusion pumps may also be suitable for use in accordance with the present invention. Examples of such devices include those described, for example, in U.S. Patents 4,692,147; 4,360,019; 4,487,603; 4,360,019; 4,725,852; 5,820,589; 5,643,207; 6,198,966 and others. In general, delivery of the compound described herein can be accomplished using any of a variety of reusable pump systems. The pumps provide continuous, controlled release over time. In some embodiments, the compound disclosed herein is in a liquid formulation in a drug-impermeable reservoir and is continuously delivered to an individual.

In one embodiment, the drug delivery system is at least partially an implantable device. The implantable device can be implanted at any suitable implantation site using methods and devices well known in the art. An implantation site is an area of the subject's body into which the drug delivery device is inserted and positioned. Implantation sites include, but are not limited to, subdermal, subcutaneous, intramuscular, or other suitable areas of the subject's body. Subcutaneous implantation sites are used in some embodiments of the invention due to the ease of implantation and removal of the drug delivery device.

Drug release devices suitable for use in the invention may be based on any of a variety of polymeric mechanisms of activity, such as, for example, poly(glycolide-co-lactide) (PGLA), which are commercially available from a number of suppliers, such as BioDegmer and Sigma-Aldrich. For example, a drug release device may be based on a diffusion system, a convective system, or an erosion system (an erosion-based system). For example, a drug release device may be an electrochemical pump, an osmotic pump, an electroosmotic pump, a vapor-pressure pump, or an osmotically disruptive matrix, such as when the drug is incorporated into a polymer (e.g., PGLA), and the polymer provides for the release of the drug formulation simultaneously with the degradation of the drug-impregnated polymeric material (e.g., a biodegradable drug-impregnated polymeric material). In other embodiments, the drug release device is based on an electrodiffusion system, an electrolytic pump, an effervescent pump, a piezoelectric pump, a hydrolytic system, etc.

Drug release devices based on a mechanical or electromechanical infusion pump may also be suitable for use in accordance with the present invention. Examples of such devices include those described, for example, in U.S. Patents Nos. 4,692,147; 4,360,019; 4,487,603; 4,360,019; 4,725,852 and others. In general, the subject method of treatment can be carried out using any of a variety of reusable disposable pump systems. Pumps and other convective systems can be used, as a rule, due to their more stable and controlled release over time. Osmotic pumps are used in some embodiments because of their combined advantages of more stable controlled release and relatively small size (see, e.g., PCT Published International Application WO 97/27840 and U.S. Patent Nos. 5,985,305 and 5,728,396). Typical osmotic devices suitable for use in the invention include, but are not limited to, those described in U.S. Patent Nos. 3,760,984; 3,845,770; 3,916,899; 3,923,426; 3,987,790; 3,995,631; 3,916,899; 4,016,880; 4,036,228; 4,111,202; 4,111,203; 4 203 440; 4 203 442; 4 210 139; 4 327 725; 4 627 850; 4 865 845; 5 057 318; 5 059 423; 5 112 614; 5 137 727; 5 234 692; 5 234 693; 5 728 396 and others. Another typical device that can be adapted for the present invention is a synchronized infusion pump (Medtronic).

In some embodiments, the drug delivery device is an implantable device. The drug delivery device can be implanted at any suitable implantation site using methods and devices well known in the art. As described herein, an implantation site is a region of the subject's body into which the drug delivery device is inserted and positioned. Implantation sites include, but are not limited to, subdermal, subcutaneous, intramuscular, or other suitable regions of the subject's body.

Suitable carrier excipients for the compound described herein include, for example, water, saline, dextrose, glycerol, ethanol, and the like, as well as combinations thereof. In addition, if desired, the carrier may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, or pH maintaining agents. Methods for preparing such dosage forms are known or will become apparent to those skilled in the art upon consideration of the present invention. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 17th edition, 1985. The administered composition or formulation will in any case contain an amount of the compound sufficient to achieve the desired condition of the subject being treated.

Compositions of the present invention include those containing a sustained-release or controlled-release matrix. Furthermore, embodiments of the present invention can be used in combination with other therapies that utilize sustained-release formulations. As used herein, the term "sustained-release matrix" is a matrix composed of materials, typically polymers, that are degradable by enzymatic or acid hydrolysis, or by dissolution. After administration to the body, the matrix is exposed to enzymes and body fluids. The sustained release matrix is preferably selected from biocompatible materials such as liposomes, polylactides (polylactic acid), polyglycolide (glycolic acid polymer), polylactide co-glycolide (copolymers of lactic and glycolic acids), polyanhydrides, poly(ortho)esters, polypeptides, hyaluronic acid, collagen, chondroitin sulfate, carboxylic acids, fatty acids, phospholipids, polysaccharides, nucleic acids, polyamino acids, amino acids such as phenylalanine, tyrosine, isoleucine, polynucleotides, polyvinylpropylene, polyvinylpyrrolidone, and silicone. Illustrative biodegradable matrices include polylactide matrix, polyglycolide matrix, and polylactide co-glycolide matrix (copolymers of lactic and glycolic acids).

In another embodiment, the polypeptide, antibody, or fragment thereof (as well as combination compositions) are delivered in a controlled release system. For example, the compound disclosed herein can be administered by intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other modes of administration. In one embodiment, a pump can be used (Sefton (1987) CRC Crit. Ref. Biomed. Eng. 14:201; Buchwald et al. (1980) Surgery 88:507; Saudek et al. (1989) N. Engl. J. Med. 321:574). In another embodiment, polymeric materials are used. In yet another embodiment, the controlled release system is located near the therapeutic target, i.e., the liver, so that only a portion of the systemic dose is required. In another embodiment, the controlled-release system is positioned close to the therapeutic target, so only a portion of the systemic agent is required. Other controlled-release systems are discussed in the review by Langer, 1990, Science 249: 1527-1533.

In another embodiment, the compositions of the present invention (as well as combination compositions, individually or together) include compositions formed by impregnating absorbent materials, such as sutures, bandages, and gauze, with the inhibitory agent described herein, or applying it to the surface of solid-phase materials, such as surgical staples, zippers, and catheters, for delivery of the compositions. Other delivery systems of this type will be apparent to those skilled in the art in light of the present invention.

The present invention provides methods and compositions for administering one or more interfering agents to a host (e.g., a human) for the treatment of a microbial infection. In various embodiments, the methods disclosed herein encompass virtually any available method and route suitable for drug delivery, including in vivo and ex vivo methods, as well as systemic and local routes of administration.

Screening tests

The present invention provides methods for screening equivalent agents, such as equivalent monoclonal antibodies to a polyclonal antibody as described herein, and various agents that modulate the activity of the active agents and pharmaceutical compositions disclosed herein, or the function of a polypeptide or peptide product encoded by a polynucleotide disclosed herein. For the purposes of the present invention, an "agent" is meant to include, but is not limited to, a biological or chemical compound, such as a simple or complex organic or inorganic molecule (referred to herein as a small molecule, such as a nucleic acid), a peptide, a protein (e.g., an antibody), an antisense polynucleotide), or a ribozyme. A wide range of compounds can be synthesized, for example, polymers such as polypeptides and polynucleotides, as well as synthetic organic compounds based on various core structures, and they are also encompassed by the term "agent." In addition, compounds for screening can be obtained from various natural sources, such as plant or animal extracts, etc. It should be understood that, although not always explicitly stated, the agent is used alone or in combination with another agent having the same or different biological activity as the agents identified by screening according to the invention.

One embodiment is a method for screening agents capable of interacting with, binding to, or inhibiting DNA-DNABII interactions (e.g., IHF or HU). Accordingly, the present invention allows for the use of virtual design methods, also known as automated design or in silico modeling, to create, select, and synthesize agents capable of interacting with, binding to, or inhibiting DNA-DNABII interactions (e.g., IHF or HU). In turn, candidate agents may be effective in the treatment of biofilms and associated diseases or conditions (medical, industrial, or veterinary), as described herein. Thus, the present invention also provides agents identified or designed using in silico methods.

A candidate agent is found to be capable of binding to DNA and/or DNABII protein if the desired interaction is detected between the candidate agent and one or both of the DNA or DNABII protein compounds. The interaction may be quantitative, for example, by the strength of interaction and/or the number of interaction sites, or qualitative, for example, by interaction or absence of interaction. The result of the method may accordingly be quantitative or qualitative. Thus, in one aspect, the present invention also provides a method for identifying an agent that does not inhibit the interaction or, alternatively, enhances the interaction between DNA and protein.

The potential inhibitory or binding effect (i.e., interaction or association) of an agent, such as a small molecule, can be analyzed prior to its actual synthesis and testing using computer modeling methods. If the theoretical structure of a given compound suggests insufficient interaction and association between it and microbial DNA in a biofilm and/or the DNABII protein, then synthesis and testing of the agent can be avoided. However, if computer modeling indicates a strong interaction, the agent can then be synthesized and tested for its ability to bind or inhibit the interaction using various methods, such as in vitro or in vivo experiments. Methods for testing the ability of an agent to inhibit or titrate biofilm, alone or in combination with another agent, are disclosed herein. This avoids the synthesis of nonfunctional agents and compounds.

A skilled artisan can use various methods to screen chemical or biological entities or fragments for their ability to bind DNABII or microbial DNA, and more specifically, specific binding sites. Selected fragments or chemical compounds can then be arranged in various orientations or docked to a specific DNA or DNABII polypeptide binding site. Docking can be performed using software such as QUANTA and SYBYL, followed by energy minimization and molecular dynamics with standard molecular mechanical force fields such as CHARMM and AMBER.

Commercial computer programs are also available for in silico design. Examples include, but are not limited to, GRID (University of Oxford, Oxford, UK), MCSS (Molecular Simulations, Burlington, MA), AUTODOCK (The Scripps Research Institute, La Jolla, CA), DOCK (University of California, San Francisco, CA), GLIDE (Schrodinger Inc.), FlexX (Tripos Inc.), and GOLD (Cambridge Crystallographic Data Centre).

After designing or selecting an agent or compound using the above methods, the efficiency with which that agent or compound can bind to one another can be tested and optimized using computational evaluation. For example, an effective DNABII fragment might exhibit a relatively small difference in energy between its bound and free states (i.e., a low binding strain energy).

A designed or selected compound can be further optimized computationally so that, when bound, it may not necessarily lose its repulsive electrostatic interaction with the target protein. Such non-complementary (e.g., electrostatic) interactions include repulsive charge-charge, dipole-dipole, and charge-dipole interactions. Specifically, the sum of all electrostatic interactions between the agent and DNABII and/or microbial DNA in the biofilm, when the agent or compound is bound to any agent, does not necessarily make a neutral or favorable contribution to the binding enthalpy.

Computer software is also available in the art for estimating the strain energy of a compound and electrostatic interaction. Examples include, but are not limited to, Gaussian 92 [Gaussian, Inc., Pittsburgh, PA]; AMBER [University of California, San Francisco]; Quanta/CHARMM [Molecular Simulations, Inc., Burlington, MA]; and Insight II/Discover [Biosysm Technologies Inc., San Diego, CA].

Once the binding agent has been optimally selected or designed as described above, substitutions of some of its atoms or side groups can be made to improve or modify its binding properties. Typically, the initial substitutions are conservative, meaning the substitution group will have approximately the same size, shape, hydrophobicity, and charge as the original group. It should, of course, be understood that components known in the art to alter conformation should be avoided. Such substituted chemical compounds can then be analyzed for their binding efficiency to the DNABII protein and/or microbial DNA in the biofilm using the same computational methods detailed above.

Some embodiments of the invention relate to a method for screening small molecules capable of interacting with a protein or polynucleotide disclosed herein. For the purposes of the present invention, "small molecules" are low molecular weight (LMW) molecules that, in one embodiment, are capable of binding to a protein of interest, thereby altering the function of the protein. In some embodiments, the molecular weight of the small molecule is 1000 or less. Methods for screening small molecules capable of altering protein function are known in the art. For example, You et al. discuss a miniature array assay for detecting small molecule-protein interactions in cells. (1997) Chem. Biol. Biol.

To implement the in vitro screening method, a suitable cell culture or tissue culture is first obtained, infected with the microorganism to be treated. The cells are cultured under appropriate conditions (temperature, growth or culture medium, and gas ( _CO2 )) and for an appropriate period of time to achieve exponential proliferation without density-dependent limitations. It is also desirable to maintain an additional, separate, uninfected cell culture as a control.

As will be apparent to one skilled in the art, suitable cells can be cultured in microtiter plates and multiple agents can be analyzed simultaneously, observing genotypic changes, phenotypic changes, or a decrease in microbial titer.

When the agent is a compound other than DNA or RNA, such as a small molecule as described above, the agent can be added directly to the cell culture or added to the culture medium. As will be apparent to those skilled in the art, an "effective" substance, which can be determined empirically, must be added.

When the agent is an antibody or antigen-binding fragment, the agent can be contacted or incubated with the target antigen and the polyclonal antibody, as described herein, under competitive ELISA conditions. Such methods are known to those skilled in the art.

Assays can also be performed on a subject. When the subject is an animal such as a rat, chinchilla, mouse, or monkey, the method provides a convenient animal model system that can be used before clinical testing of the agent in humans. In this system, the candidate agent is considered a potential therapeutic agent if the symptoms of the disease or microbial infection are reduced or eliminated compared to an untreated animal with the same infection. It can also be useful to have a separate group of negative control cells or animals that are healthy and untreated, providing a basis for comparison.

The agents and compositions can be used in the manufacture of medicinal products and for the treatment of humans and other animals by administration in accordance with generally accepted procedures, for example, as an active ingredient in pharmaceutical compositions.

The compositions and related methods of the present invention can be used in combination with other therapies. These include, but are not limited to, the administration of DNAse enzymes, antibiotics, antimicrobials, anti-infectives, antifungals, antiparasitic agents, antiviral agents, or other antibodies.

In some embodiments, the methods and compositions include a deoxyribonuclease (DNase) enzyme that acts synergistically with an anti-DNABII antibody. DNase is any enzyme that catalyzes the cleavage of phosphodiester bonds in the DNA backbone. Three non-limiting examples of DNase enzymes that are known to target not only cruciform structures but also various secondary structures of DNA include DNase I, T4 EndoVII, T7 Endo I, RuvABC, and RusA. In some embodiments, the effective amount of anti-DNABII antibody required to destabilize the biofilm is reduced when combined with DNase. When administered in vitro, DNase can be added directly to the assay system or to a suitable buffer known to stabilize the enzyme. The effective single dose of DNase and assay conditions can vary and can be optimized according to procedures known in the art.

In other embodiments, the methods and compositions can be combined with antibiotics and/or antimicrobials. Antimicrobials are substances that kill or inhibit the growth of microorganisms, such as bacteria, fungi, or protozoa. Although biofilms are generally resistant to antibiotics, the compositions and methods described herein can be used to sensitize an infection, including a biofilm, to traditional therapeutic treatments for infections. In other embodiments, the use of antibiotics or antimicrobials in combination with the methods and compositions described herein allows for a reduction in the effective amount of the antimicrobial and/or biofilm-disrupting agent. Some non-limiting examples of antimicrobials and antibiotics useful in combination with the methods of the present invention include amoxicillin, amoxicillin-clavulanate, cefdinir, azithromycin, and sulfamethoxazole-trimethoprim. A therapeutically effective dose of an antimicrobial and/or antibiotic in combination with a biofilm-disrupting agent can be readily determined using conventional methods. In some embodiments, the dose of the antimicrobial agent in combination with the biofilm-disrupting agent represents a median effective dose that has been shown to be effective against other bacterial infections, such as bacterial infections in which the etiology of the infection does not involve biofilm. In other embodiments, the dose is 0.1, 0.15, 0.2, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.8, 0.85, 0.9, 0.95, 1.1, 1.2, 1, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3.0, or 5 times the average effective dose. The antibiotic or antimicrobial agent can be added before, simultaneously with, or after the addition of the anti-DNABII antibody.

In other embodiments, the methods and compositions can be combined with antibodies that treat a bacterial infection. One example of an antibody used in combination with the methods and compositions described herein is an antibody directed against an unrelated outer membrane protein (i.e., OMP P5). Treatment with this antibody alone does not reduce biofilm volume in vitro. Combination therapy with this antibody and a biofilm-disrupting agent results in a greater effect than that which can be achieved using either reagent alone at the same concentration. Other antibodies that can provide a synergistic effect when combined with a biofilm-disrupting agent or a biofilm-reducing method include anti-rsPilA, anti-OMP26, anti-OMP P2, and anti-all OMP types.

The compositions and methods described herein can be used to sensitize a bacterial infection involving biofilm formation to conventional therapeutic approaches that are effective in treating bacterial infections without biofilm, but otherwise ineffective in treating bacterial infections involving biofilms. In other embodiments, the compositions and methods described herein can be used in combination with therapeutic approaches that are effective in treating bacterial infections associated with biofilm, but the combination of such additional therapy and a biofilm-disrupting agent or method produces a synergistic effect, such that the effective dose of either the biofilm-disrupting agent or the additional therapeutic agent can be reduced. In other cases, the combination of such additional therapy and a biofilm-disrupting agent or method produces a synergistic effect, such that treatment is intensified. Intensification of treatment may be evidenced by a shorter time required to treat the infection.

The additional therapeutic treatment may be carried out before, simultaneously with or after the methods or compositions used to destroy biofilms, and the agent used therein may be contained in the same composition/preparation or be presented as a separate preparation/composition.

Sets

Also disclosed are kits containing agents and instructions necessary for performing the methods described herein in vitro and in vivo. Accordingly, the description provides kits for performing these methods, which may include an antibody, an antibody fragment, a polypeptide, a polynucleotide, a vector, or a host cell, as well as instructions for performing the methods disclosed herein, such as collecting tissue and/or performing screening and/or analyzing the results, and/or administering an effective amount of an antibody, an antibody fragment, a polypeptide, a polynucleotide, a vector, or a host cell, as defined herein. They can be used alone or in combination with other suitable antimicrobial agents.

For example, the kit may comprise or alternatively consist essentially of or additionally consist of any compound selected from one or more of the agents mentioned above, such as an antibody, antibody fragment, polypeptide, polynucleotide, vector, or host cell, and instructions for use. The kit may further comprise one or more adjuvants, an antigenic peptide, or an antimicrobial agent. Examples of carriers include a liquid carrier, a pharmaceutically acceptable carrier, a solid-phase carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable polymer, a liposome, a micelle, an implant, a stent, a paste, a gel, a dental implant, or a medical implant.

The following examples are intended to illustrate and not to limit the embodiments disclosed in this application.

Experimental part

Experiment #1

Antibodies were generated from two IHF(NTHI) head chimeric peptides and an IHF(NTHI) tail chimeric peptide, in addition to antibodies from two mouse hybridoma cell lines: a clone producing antibodies to the IHF(NTHI) head chimeric peptide and a clone producing antibodies to the IHF(NTHI) tail chimeric peptide. Heavy and light chains were sequenced. A chimeric parenteral antibody, consisting of the murine variable region sequences of two individual antibodies, was fused to the framework region of the human IgG1 constant region. The specificity of each clone to the corresponding synthetic peptide was confirmed by ELISA. A fully human antibody was generated, and nine antibody variants to the head chimeric peptide and nine antibody variants to the tail chimeric peptide were obtained, and the specificity of each clone for its respective target was confirmed by ELISA (see Experiment No. 2). The avidity and affinity of each antibody were determined.

Experiment #2

To assess the binding of the humanized variants to the corresponding peptides, a direct binding ELISA was performed. The head or tail chimeric peptides were coated on a 96-well plate at a concentration of 2 μg/mL, and an 8-fold antibody dilution series (starting concentration 50 μg/mL, dilution 1:3) was added. HRP-conjugated anti-human IgG Fc antibody (1:7000, Jackson Immuno Research, 109-035-098) was used as a secondary detection antibody. A standard direct ELISA protocol was followed, and the optical density at 450 nm was measured using a microplate reader. _EC50 values were calculated, and the humanized variants were determined to have comparable binding. See Figs. 2A–2D and Tables 1 and 2 below.

Table 1 Results of ELISA using humanized antibodies against the head portion.

Heavy chain Light chain ELISA #1:
peptide as a target (titration curves do not reach equilibrium)
EC ₅₀ (ng/ml) ELISA #2:
peptide as target
(titration curves reach equilibrium)
EC ₅₀ (ng/ml) SEQ ID NO: 1 SEQ ID NO: 7 60.0 70.4 SEQ ID NO: 1 SEQ ID NO: 8 66.0 64.4 SEQ ID NO: 1 SEQ ID NO: 9 32.0 55.0 SEQ ID NO: 2 SEQ ID NO: 7 49.0 62.0 SEQ ID NO: 2 SEQ ID NO: 8 55.0 40.2 SEQ ID NO: 2 SEQ ID NO: 9 27.0 45.3 SEQ ID NO: 3 SEQ ID NO: 7 75.0 85.8 SEQ ID NO: 3 SEQ ID NO: 8 66.0 93.4 SEQ ID NO: 3 SEQ ID NO: 9 35.0 111.1

Table 2. Results of ELISA using humanized antibodies against the tail region.

Heavy chain Light chain ELISA #1:
peptide as a target (titration curves do not reach equilibrium)
EC ₅₀ (ng/ml) ELISA #2:
peptide as target
(titration curves reach equilibrium)
EC ₅₀ (ng/ml) SEQ ID NO: 4 SEQ ID NO: 10 10.0 42.7 SEQ ID NO: 4 SEQ ID NO: 11 11.0 40.8 SEQ ID NO: 4 SEQ ID NO: 12 12.0 40.4 SEQ ID NO: 5 SEQ ID NO: 10 7.0 36.7 SEQ ID NO: 5 SEQ ID NO: 11 14.0 33.9 SEQ ID NO: 5 SEQ ID NO: 12 15.0 42.9 SEQ ID NO: 6 SEQ ID NO: 10 13.0 27.4 SEQ ID NO: 6 SEQ ID NO: 11 11.0 28.0 SEQ ID NO: 6 SEQ ID NO: 12 8.0 29.4

Surface plasmon resonance (SPR) was also used to determine the affinity of humanized MAbs for the head or tail chimeric peptide (i.e., the target peptide) and for native IHF _NTHI . SPR was performed using a Biacore 3000 instrument (GE Healthcare Life Sciences). All experiments were carried out at 25°C with 10 mM HEPES (pH 7.4) - 150 mM NaCl - 3 mM EDTA - 0.005% surfactant P20 (HBS-EP; GE Healthcare) as the running buffer.

In one experimental setup, surface plasmon resonance (SPR) was performed as follows: a reagent-grade CM5 sensor chip was used to immobilize the IHF _NTHI head or IHF _NTHI tail chimeric peptide onto separate flow cells using amino-coupling chemistry. Each monoclonal antibody was diluted twofold from 100 nM to 3.1 nM in HBS-EP + NSB buffer and injected across the sensor chip surface at a flow rate of 30 μL/min, with a 2-min injection cycle and a 2-min dissociation cycle. A negative control injection cycle with buffer only was also included to detect nonspecific binding. Association and dissociation constants were determined using BiaEvaluation.

In one experimental setting, using amino-linked chemistry and a flow rate of 5 μL/min, a humanized monoclonal antibody was immobilized on the flow cells of the CM5 sensor chip at 4000 resonance units (RU) for target peptide binding analysis or 2000 RU for native IHF _NTHI binding assessment. The target peptide or native IHF _NTHI was then suspended in HBS-EP plus NSB reducing agent, and serial two-fold dilutions from 100 nM to 3.1 nM, including a buffer-only sample, were injected through the antibody-linked surface at a flow rate of 30 μL/min, an injection time of 5 min, and a dissociation time of 5 min using the KINJECT command. BiaEvaluation software was used to fit the sensorgram curves, subtract the buffer-only injection cycle, and determine the KD values. The results are shown in Tables 3 and 4 below.

Table 3. SPR results using humanized anti-head antibodies.

Heavy chain Light chain SPR: Peptide immobilized, HuMab as analyte SPR: IHF _NTHI immobilized, HuMab as analyte k _on (M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) k _on
(M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) SEQ ID NO: 1 SEQ ID NO: 7 3.1E+04 2.2E-04 7.0 5.4E+03 9.0E-05 17.0 SEQ ID NO: 1 SEQ ID NO: 8 4.2E+04 1.1E-05 0.3 1.5E+05 6.0E-03 40.0 SEQ ID NO: 1 SEQ ID NO: 9 4.0E+04 1.3E-05 0.3 6.5E+06 1.6E-01 24.0 SEQ ID NO: 2 SEQ ID NO: 7 1.8E+04 2.6E-03 144.0 3.1E+01 1.4E-03 4400.0 SEQ ID NO: 2 SEQ ID NO: 8 2.2E+04 2.8E-04 13.0 4.2E+02 1.5E-04 354.0 SEQ ID NO: 2 SEQ ID NO: 9 3.7E+03 1.2E-05 4.0 4.6E+00 1.7E-03 125.0 SEQ ID NO: 3 SEQ ID NO: 7 4.8E+04 4.0E-04 8.0 1.4E+02 1.1E-04 825.0 SEQ ID NO: 3 SEQ ID NO: 8 5.1E+04 3.5E-04 7.0 1.8E+03 2.1E-04 115.0 SEQ ID NO: 3 SEQ ID NO: 9 7.9E+04 2.1E-04 3.0 4.6E+03 1.6E-02 3400.0

Table 4 SPR results using humanized anti-tail antibodies.

Heavy chain Light chain SPR: Peptide immobilized, HuMab as analyte k _on (M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) SEQ ID NO: 4 SEQ ID NO: 10 1.5E+05 6.4E-04 4.3 SEQ ID NO: 4 SEQ ID NO: 11 1.4E+05 6.8E-04 4.9 SEQ ID NO: 4 SEQ ID NO: 12 1.1E+05 5.6E-04 5.1 SEQ ID NO: 5 SEQ ID NO: 10 1.3E+05 8.2E-04 6.1 SEQ ID NO: 5 SEQ ID NO: 11 1.2E+05 7.6E-04 6.2 SEQ ID NO: 5 SEQ ID NO: 12 8.5E+04 7.7E-04 9.1 SEQ ID NO: 6 SEQ ID NO: 10 1.8E+05 4.6E-04 2.5 SEQ ID NO: 6 SEQ ID NO: 11 1.6E+05 1.6E-04 1.0 SEQ ID NO: 6 SEQ ID NO: 12 2.5E+05 6.0E-04 2.4

In another experimental setup, using amine-coupled chemistry and a flow rate of 5 μl/min, the target peptide or native IHF _NTHI was immobilized in the flow cells of the CM5 sensor chip at 4000 resonance units (RU) or 2000 RU, respectively, for humanized monoclonal antibody binding analysis. Humanized monoclonal antibodies were then suspended in HBS-EP plus NSB reducing agent, and serial two-fold dilutions from 100 nM to 3.1 nM, including a buffer-only sample, were injected through the peptide/IHF _NTHI- coupled surface at a flow rate of 30 μl/min, an injection time of 5 min, and a dissociation time of 5 min using the KINJECT command. BiaEvaluation software was used to fit the sensorgram curves, subtract the buffer-only injection cycle, and determine the KD values. The results are shown in Tables 5 and 6 below.

Table 5. SPR results using humanized anti-head antibodies.

Heavy chain Light chain SPR: HuMab immobilized, peptide as analyte SPR: HuMab immobilized, IHF _NTHI as analyte k _on (M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) k _on
(M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) SEQ ID NO: 1 SEQ ID NO: 7 4.5E+03 3.4E-04 76.0 1.4E+03 1.4E-05 10.0 SEQ ID NO: 1 SEQ ID NO: 8 1.7E+04 3.7E-04 21.0 1.4E+06 3.2E-04 0.2 SEQ ID NO: 1 SEQ ID NO: 9 2.5E+03 3.8E-04 152.0 6.3E+05 3.7E-04 0.6 SEQ ID NO: 2 SEQ ID NO: 7 2.8E+02 1.5E-04 527.0 6.4E+05 4.0E-04 0.6 SEQ ID NO: 2 SEQ ID NO: 8 3.0E+02 7.7E-05 257.0 7.0E+04 5.9E-04 9.0 SEQ ID NO: 2 SEQ ID NO: 9 3.5E+03 7.4E-05 23.0 7.2E+03 4.5E-04 62.0 SEQ ID NO: 3 SEQ ID NO: 7 5.4E+04 1.6E-03 29.0 6.6E+04 4.0E-04 6.0 SEQ ID NO: 3 SEQ ID NO: 8 9.5E+04 1.9E-03 20.0 7.5E+03 1.8E-03 245.0 SEQ ID NO: 3 SEQ ID NO: 9 9.5E+03 1.9E-03 200.0 1.7E+03 2.7E-03 1600.0

Table 6. SPR results using humanized anti-tail antibodies.

Heavy chain Light chain SPR: HuMab immobilized, peptide as analyte SPR: HuMab immobilized, IHF _NTHI as analyte k _on (M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) k _on
(M ^-1 s ^-1 ) k _off (s ^-1 ) K _D (nM) SEQ ID NO: 4 SEQ ID NO: 10 1.1E+04 3.3E-04 29.6 7.7E+04 4.3E-04 33.1 SEQ ID NO: 4 SEQ ID NO: 11 4.3E+02 2.7E-04 62.2 1.0E+05 5.3E-04 53.0 SEQ ID NO: 4 SEQ ID NO: 12 NS NS NS NS NS NS SEQ ID NO: 5 SEQ ID NO: 10 4.8E+03 2.9E-04 60.0 1.6E+05 5.7E-04 93.4 SEQ ID NO: 5 SEQ ID NO: 11 5.7E+03 2.7E-04 47.0 1.4E+05 6.1E-04 82.4 SEQ ID NO: 5 SEQ ID NO: 12 2.0E+04 4.4E-04 23.0 3.3E+04 1.2E-03 39.0 SEQ ID NO: 6 SEQ ID NO: 10 1.8E+04 1.3E-03 72.0 2.5E+04 7.3E-04 18.0 SEQ ID NO: 6 SEQ ID NO: 11 3.3E+04 1.3E-03 67.0 4.6E+04 6.8E-04 31.0 SEQ ID NO: 6 SEQ ID NO: 12 1.6E+05 2.1E-03 13.0 7.7E+03 1.1E-03 8.5

* NS = not done

Experiment #3

This experiment describes an in vitro model for reversing established biofilm on an 8-well microscope slide. The following materials were used in this experiment: chocolate agar; sBHI (BHI with 2 mg heme/mL and 2 mg b-NAD/mL); 8-well microscope slides (Nunc* Lab-Tek* Fisher cat. #12-565-18); sterile 0.9% saline; LIVE/DEAD BacLight Bacterial Viability Assay Kit (Fisher cat. #NC9439023), and formalin.

Colonies of NTHI 86-028NP were picked from an overnight culture on chocolate agar and suspended in brain heart infusion broth supplemented with 2 μg of β-NAD and heme per ml of medium (sBHI). The optical density at 490 nm was then adjusted to 0.65, and the culture was diluted 1:6 in sBHI before incubation at 37°C with 5% CO2 for 3 hours without stirring. The culture was then diluted 1:2500 in fresh sBHI, and 200 μl aliquots of the suspension were added to each well of an 8-well microscope slide. The slide was then incubated at 37°C with 5% CO2 for 3 hours without stirring. After 16 hours, 200 μl of fresh sBHI were added to each well and the slide was incubated for an additional 8 hours. At this point, the medium was aspirated from each well and monoclonal antibodies were added at 5 μg per well. The biofilms were incubated for an additional 16 hours. Biofilms were then washed and stained with FM1-43FX bacterial cell membrane dye (Invitrogen) and fixed overnight at 4°C in 16% paraformaldehyde, 2.5% glutaraldehyde, 4.0% acetic acid in 0.1 M phosphate buffer (pH 7.4). The fixative was aspirated. 200 μl of 0.9% Hanks' balanced salt solution was added to each well before viewing the biofilms on a Zeiss 800 Meta-laser scanning confocal microscope. Images were compiled using Zeiss Zen Blue software, and biofilm biomass was calculated using COMSTAT2 software. See Fig. 3 and Tables 7–8 below.

Table 7. In vitro biofilm destruction occurs when using humanized antibodies against the head portion.

Heavy chain Light chain Biofilm destruction
in vitro
(% change in biomass) SEQ ID NO: 1 SEQ ID NO: 7 ↓87 SEQ ID NO: 1 SEQ ID NO: 8 ↓80 SEQ ID NO: 1 SEQ ID NO: 9 ↓90 SEQ ID NO: 2 SEQ ID NO: 7 ↓50 SEQ ID NO: 2 SEQ ID NO: 8 ↓60 SEQ ID NO: 2 SEQ ID NO: 9 ↓70 SEQ ID NO: 3 SEQ ID NO: 7 ↓87 SEQ ID NO: 3 SEQ ID NO: 8 ↓80 SEQ ID NO: 3 SEQ ID NO: 9 ↓67

↓ indicates a decrease in biomass; while ↑ indicates an increase in biomass.

Table 8. In vitro biofilm disruption using humanized anti-tail antibodies.

Heavy chain Light chain Biofilm destruction
in vitro
(% change in biomass) SEQ ID NO: 4 SEQ ID NO: 10 ↓6 SEQ ID NO: 4 SEQ ID NO: 11 ↑7 SEQ ID NO: 4 SEQ ID NO: 12 ↑8 SEQ ID NO: 5 SEQ ID NO: 10 ↓4 SEQ ID NO: 5 SEQ ID NO: 11 ↑4 SEQ ID NO: 5 SEQ ID NO: 12 0 SEQ ID NO: 6 SEQ ID NO: 10 ↓6 SEQ ID NO: 6 SEQ ID NO: 11 ↓3 SEQ ID NO: 6 SEQ ID NO: 12 ↑3

↓ indicates a decrease in biomass; while ↑ indicates an increase in biomass.

Experiment #4

In addition to Experiments 1-3 above, the following study is described demonstrating targeting of the bacterial DNABII protein with a chimeric peptide immunogen or humanized monoclonal antibody to prevent or treat non-biofilm-mediated infections.

Research overview

First, as a therapeutic approach, the applicant utilized intact IgG or Fab fragments against a chimeric peptide immunogen designed to target protective epitopes in the DNA-binding head domains of a host integration factor to disrupt established biofilms in vitro and mediate disease cure in vivo. Second, the applicant conducted prophylactic active immunization with the chimeric peptide to induce antibody production that blocks biofilm formation and disease progression in a viral-bacterial superinfection model. Furthermore, in preparation for clinical application, the applicant humanized the monoclonal antibody against the chimeric peptide immunogen, then characterized and confirmed its therapeutic efficacy.

The applicant demonstrated the efficacy of each approach in two well-established preclinical models of otitis media caused by the prevalent atypical respiratory pathogen Haemophilus influenzae, a common biofilm disease. Taken together, the applicants identified two approaches with significant efficacy and potential for widespread use in the treatment of diseases with a biofilm component.

The objectives of Experiment 4 were to (1) test the preclinical therapeutic potential of the IgG antigen-binding domains (Fab fragments) of a monoclonal antibody directed against a DNA-BII-directed head peptide for the treatment of a well-characterized biofilm disease, NTHI-associated otitis media, and (2) test the same head peptide as an immunogen for its ability to induce antibodies that would prevent biofilm formation and disease when used as a vaccine antigen in active immunization regimens as an adjunctive prophylactic strategy. Applicant initially evaluated Fab or intact IgG for head peptide-mediated disruption of bacterial biofilms in vitro. The applicant then used two well-established chinchilla models of the common biofilm disease, otitis media, caused by NTHI (one based on direct bacterial infection of the middle ear and the other on a viral-bacterial superinfection model of ascending OM) and demonstrated the preclinical efficacy of both therapeutic and prophylactic strategies targeting DNABII. Finally, the applicant humanized one DNABII-targeted monoclonal antibody and confirmed its in vitro activity and preclinical therapeutic efficacy.

In vitro biofilm disruption assays were repeated three times on separate days. For in vivo experiments, chinchillas were randomly divided into groups based on body weight; both male and female animals were included. To examine the disruption of NTHI middle ear biofilms induced by mouse anti-β-head Fabs, anti-β-tail Fabs, or isotype control Fabs, three or four animals were included in each group. To test biofilm disruption induced by Fabs from rabbit polyclonal IgG serum against the head chimeric peptide, IgG serum against the tail chimeric peptide, or naive rabbit IgG serum, groups of six chinchillas each were created. The efficacy of HuTipMabs compared with HuTailMabs or saline was evaluated in six animals per group. Neither animals nor samples were excluded from the study. The relative quantitative value of mucosal biofilm remaining in the middle ear after treatment was assessed by 6-8 participants who were not participating in the study and were unaware of the treatment received. For each study assessment, each middle ear was analyzed independently.

Mouse monoclonal antibodies, rabbit polyclonal antibodies and humanized monoclonal antibodies

Mouse monoclonal antibodies against the β-head (clone 12E6.F8.D12.D5, mIhfB4NTHI) or β-tail (clone 7A4.E4.G11, IhfB2NTHI) domains of IHF _NTHI were purified from cell culture supernatants as described by LA Novotny et al. (2016) EBioMedicine 10: 33–44. Mouse IgG1κ isotype control antibody (clone P3.6.2.8.1; eBioscience cat. #16-4714-82, RRID: AB_470161) served as a negative control. Fab fragments were then generated by papain digestion with the Mouse IgG1 Fab Kit and the F(ab') ₂ Preparation Kit (Pierce) according to the instructions. Polyclonal rabbit antibodies against the head chimeric peptide and tail chimeric peptide were obtained from Rockland Immunochemical, Inc. (LA Novotny et al. (2014) Mol. Microbiol 93:1246–1258). Naive rabbit serum served as a negative control. IgG was enriched from each rabbit's serum by passing through rProtein A Protein G GraviTrap columns (GE Healthcare) according to the manufacturer's instructions. Fabs were then generated using the Pierce Fab Fragment Generation Kit. Cleavage of intact mouse or rabbit IgG to generate Fab was confirmed by SDS-PAGE stained with Coomassie Fluor™ Orange Protein Gel (ThermoFisher). Humanized monoclonal antibodies against the head chimeric peptide (HuTipMab) or tail chimeric peptide (HuTailMab) were obtained from LakePharma, Inc. and are described briefly above. Antigen-binding domains were derived from a mouse monoclonal antibody directed against the head chimeric peptide (clone 1F8.C3.D11.F1) or tail chimeric peptide (clone 11E7.G11.C7). For this study, clones of humanized monoclonal antibodies TP-21949 (targeting the head chimeric peptide; HuTipMab) and TP-21958 (targeting the tail chimeric peptide; HuTailMab) were used. Bacterial endotoxin testing using the ToxinSensor Chromogenic LAL Endotoxin Kit (Genscript) was performed on all antibody lots before use.

In vitro biofilm destruction

Biofilms formed by strain NTHI 86-028NP (a minimally passaged clinical isolate from the nasopharynx of a child with chronic OM, see, for example, Bakaletz LO et al. (1988) Infect Immun 1988; 56: 331–5), strain 7169 M. catarrhalis (a minimally passaged clinical isolate from the middle ear of a child with chronic OM, see, for example, Luke NR et al (1999) Infect Immun 1999; 67: 681–7), strain 27853 P. aeruginosa, strain K56 B. cenocepacia (isolated from sputum of a patient with cystic fibrosis, see, for example, Mahenthiralingam E et al (2000) J Clin Microbiol 2000; 38: 910–3) and S. aureus strain 29213 were first cultured in an 8-well coverslip (CellVis) for 24 h and then incubated with 170 nM intact IgG or Fab fragments for an additional 16 h (S.D. Goodman et al. (2011) Mucosal Immunol 4:625–637; J.A. Jurcisek et al. (2011) J Vis Exp). In one embodiment, strain NTHI 86-028NP has been maintained at no more than 4 passages since its isolation from a child in 1986. Additionally or alternatively, strain Mcat has similarly been maintained at a very low passage number since its isolation. In another embodiment, Burkholderia bacteria were isolated from sputum of a patient with cystic fibrosis (CF). In another embodiment, a Staphylococcus aureus isolate was obtained from ATCC. The 170 nM concentration was based on previous studies in which 5 μg intact IgG per 0.2 mL volume was applied to biofilms in vitro (Goodman SD et al (2011) Mucosal Immunol 2011; 4: 625–37; Brockson ME et al (2014) Mol Microbiol 2014; 93: 1246–58; Novotny LA et al (2019) NPJ Vaccines 2019; 4: 43; Novotny LA et al (2013) PLoS One 2013; 8: e67629; and Novotny LA et al (2016) EBioMedicine 2016; 10: 33–44) to allow direct comparison of IgG- and Fab-mediated destruction. Bacteria within biofilms were then stained with FM1-43FX (ThermoFisher), fixed overnight in a solution of 16% paraformaldehyde, 4% acetic acid, and 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4), and then washed with 10 mM phosphate-buffered saline (pH 7.4). Biofilms were viewed with a Zeiss 800 scanning confocal laser microscope, images were visualized in Zeiss Zen Pro software, and biomass was determined with COMSTAT2 software (A. Heydorn et al. (2000) Microbiology 146 (pt 10): 2395–2407). In vitro biofilm disruption assays were repeated three times on separate days.

Animals

Work with chinchillas was conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals and under protocol #01304AR, which was approved by the Abigail Wexner Research Institute, Nationwide Children's Hospital Institutional Animal Care and Use Committee. Juvenile or adult chinchillas (Chinchilla lanigera; juveniles weighed 250–499 g; adults weighed 500–850 g) were obtained from Rauscher's Chinchilla Ranch, LLC. These non-specific pathogen-free, non-pedigree animals were housed in individual cages with autoclaved corn cob bedding and sterile water ad libitum. Animals were randomly assigned to groups based on body weight (as an indicator of juvenile or adult status), and both males and females were used. The experimental groups were as follows: To study the disruption of NTHI middle ear biofilms induced by mouse anti-β-head Fab, anti-β-tail Fab, or isotype control Fab, three or four animals were included in each group (mean body weight per group, 625 g). To test the biofilm disruption induced by Fab made from polyclonal serum IgG against the head chimeric peptide, serum IgG against the tail chimeric peptide, or IgG made from naive rabbit serum, groups of three chinchillas each were created (mean body weight per cohort, 505 g). The efficacy of HuTipMab compared with HuTailMab or saline was evaluated in three animals per group (mean body weight per group, 580 g). To evaluate the ability of the head chimeric peptide to induce the formation of antibodies that prevent the development of NTHI-induced OM, eight young animals per group (average body weight per group 430 g) were used.

Destruction of NTHI biofilms formed in the middle ears of chinchillas under experimental OM conditions

Both middle ears of each animal were inoculated with 1000 CFU of NTHI strain 86-028NP by direct injection to induce experimental OM. After four days, NTHI mucosal biofilms filled more than 50% of the middle ear (L.A. Novonty et al. (2011) Mucosal Immunol 4: 456–467). At this time, 342 nM Fab or humanized monoclonal antibody was inoculated into each middle ear (delivered at 0.1 ml per bulla), with identical administration 24 h later. The 342 nM concentration was based on previous studies in which 5 μg intact IgG per 0.1 ml volume was inoculated into the middle ears of chinchillas (Goodman SD et al 92011) Mucosal Immunol 2011; 4: 625–37; Novotny LA et al (2019) NPJ Vaccines 2019; 4: 43; and Novotny LA et al (2016) EBioMedicine 2016; 10: 33–44) 11, 13, 21, as well as for direct comparison of IgG- and Fab-mediated destruction in vivo. To determine the immediate treatment outcome, animals were sacrificed one day after completion of antibody therapy, images of mucosal biofilms were obtained with a Zeiss SV6 dissecting microscope, and mucosal biofilms and middle ear mucosa were then collected, homogenized, and plated on chocolate agar to quantify the bacterial load in the middle ear (S.D. Goodman et al. (2011) Mucosal Immunol 4: 625–637). In two of the studies described in this application (evaluation of biofilm disruption by polyclonal rabbit IgG Fab fragments and the efficacy of a humanized monoclonal antibody in disrupting mucosal biofilms, as well as the cytokine profile of animals treated with HuMab), a subset of animals in each group were observed for an additional seven days without additional treatment to examine whether NTHI biofilms would re-form after cessation of antibody therapy and to determine how the cytokine profile might change over time. Mucosal biofilms were collected and processed as described (S.D. Goodman et al. (2011) Mucosal Immunol 4: 625–637).

Quantitative assessment of mucosal biofilm

As an additional assessment, the amount of biofilm in each middle ear was qualitatively determined. Images of each middle ear were obtained, randomized, and assessed by six reviewers blinded to treatment using an established rubrication where 0 = no mucosal biofilm visible, 1 = <25% of middle ear occluded by mucosal biofilm, 2 = ≥25-50% occlusion, 3 = ≥50-75% occlusion, 4 = ≥75-100% occlusion (L.A. Novotny et al. (2011) Mucosal Immunol 4: 456-467). For both quantitative bacterial load determination and qualitative mucosal biofilm assessment, each middle ear was considered independent.

Quantitative determination of cytokines in middle ear fluid

To quantify cytokines in middle ear fluids (MEFs), cytometric analysis was performed with BD ^™ beads on fluids collected at the time of animal sacrifice. Using human-specific Flex BD ^™ kits, each MEF was individually assayed for IL-1β (Cat. #558279), IL-6 (Cat. #558276), IL-8 (Cat. #558277), IL-10 (Cat. #558274), IL12-p70 (Cat. #558283), IL-17A (Cat. #560383), and TNF (Cat. #560112) according to the manufacturer's instructions. The cytokine IL-13 (catalog #558450) was added to the panel used to analyze MEFs collected in the HuMabs study. MEFs from each animal were analyzed individually. Data were acquired on a BD Accuri™ C6 cytometer (BD Biosciences) and analyzed using FlowJo software (FlowJo, LLC). Cytokine concentrations in each MEF were determined using a standard curve and calculated using GraphPad Prism software.

Surface plasmon resonance

To determine the affinity of the humanized monoclonal antibody directed against the head chimeric peptide and native IHF _NTHI , surface plasmon resonance was performed using a Biacore 3000 instrument (GE Healthcare Life Sciences). All experiments were performed at 25°C and 10 mM HEPES (pH 7.4) - 150 mM NaCl - 3 mM EDTA - 0.005% surfactant P20 (HBS-EP; GE Healthcare, cat. no. BR100188) as a running buffer. Using amino-coupling chemistry and at a flow rate of 5 µl/min, a humanized monoclonal antibody specific for the head chimeric peptide was immobilized onto flow cells of the CM5 sensor chip (GE Healthcare, cat. no. BR100012) at 4000 resonance units (RU) for head chimeric peptide binding analysis or 2000 RU for native IHF _NTHI binding assessment. The head chimeric peptide or native IHF _NTHI was then suspended in HBS-EP plus NSB reducing agent (GE Healthcare, cat. #BR100691) and serial two-fold dilutions from 100 nM to 3.1 nM, including buffer-only sample, were injected through the antibody-coupled surface at a flow rate of 30 μl/min, an injection time of 5 min, and a dissociation time of 5 min using the KINJECT command. BiaEvaluation software was used to fit the sensorgram curves, subtract the buffer-only injection cycle, and determine _KD values.

Model of viral-bacterial co-infection of experimental NTHI-induced OM

An established adenovirus-NTHI polymicrobial challenge model (K. Suzuki et al. (1994) Infect Immun 62: 1710–1718) was used to evaluate the efficacy of the head chimeric peptide when delivered as a preclinical vaccine antigen and/or to evaluate the ability of the head chimeric peptide to induce antibody production that prevented the development of NTHI-induced OM. Chinchillas were randomly divided into three groups of eight animals each based on body weight and immunized by rubbing the vaccine formulations into intact skin behind each pinna (retroauricular region) (LA Novotny et al. (2017) Clin Vaccine Immunol 24). The formulations consisted of 10 μg of the head chimeric peptide or 10 μg of the tail chimeric peptide, each mixed with 10 μg of the adjuvant LT(R192G/L211A), a heat-labile enterotoxin of double mutant E. coli (dmLT; a generous gift from Dr. John Clements) (JD Clements et al. (2018) dmLT. mSphere 3). As a negative control, one group received 10 μg of dmLT only. A second identical dose was administered one week later. Two days after the second dose, the time when the maximal immune response is observed for this immunization schedule (LA Novotny et al. (2013) Vaccine 31: 3417–3426), all chinchillas were administered 1.9 x 107 TCID ₅₀ adenovirus serotype 1 by passive inhalation of droplets. Seven days later, when adenoviral damage to the respiratory tract occurs (K. Suzuki et al. (1994) Infect Immun 62: 1710–1718), all animals were challenged with 108 CFU of the NTHI 86-028NP strain by passive droplet inhalation. To confirm that all groups were equally colonized with NTHI (now residing within the nasopharynx) and thus available for ascent through the virus-weakened eustachian tube, a nasopharyngeal lavage was performed one day after the bacterial challenge. Nasopharyngeal swabs were serially diluted and plated on chocolate agar supplemented with 15 μg ampicillin/ml medium to limit the growth of normal chinchilla flora (LA Novotny et al. (2017) Clin Vaccine Immunol 24: e00563-16. This concentration of ampicillin does not affect the growth of the NTHI 86-028NP strain). NTHI colonies were counted after incubation for 24 h at 37°C.

Video otoscopy

All animals underwent video otoscopy using a Welch Allyn MacroView™ otoscope and Welch Allyn Viewer software. Each tympanic membrane was scored blindly on a predetermined scale of 0 to 4, and a middle ear with a score ≥2.0 was considered OM-positive because inflammation (erythema) and MEF were visible (L.A. Novotny et al. (2006) Vaccine 24: 4804–4811). If one middle ear scored ≥2.0 and the contralateral ear scored <2.0, the animal was considered OM-positive. To calculate vaccine efficacy, the number of OM observations during the 20-day study period was first determined and converted to a percentage of the total number of observations for each group. This value was then subtracted from the percentage calculated for the dmLT-only group. The video otoscopy was performed by a person who was unaware of the composition being administered.

Statistical analysis

Graphical results and statistical tests were performed using GraphPad Prism 8. Differences in biomass for in vitro biofilm disruption assays were determined using one-way ANOVA with multiple comparisons. Differences in cytokine levels in middle ear fluids between groups were determined using one-way ANOVA with multiple comparisons. Differences in bacterial load and mean mucosal biomass were determined using one-way ANOVA with multiple comparisons. Delay in OM onset and time to disease resolution were determined using the Mantel-Cox test.

RESULTS

Fab fragments of a mouse monoclonal antibody against the β-head portion disrupted bacterial biofilms in vitro

The applicant first conducted an in vitro coating assay in which biofilms formed by five different bacterial species were allowed to form for 24 hours before incubation with Fab fragments to confirm their potential activity. The Fab fragments were generated from a murine monoclonal antibody directed against a previously defined 15-mer immunoprotective domain in the DNA-binding "head" region of the β-subunit of IHF NTHI (IHF _NTHI , referred to herein as "anti-β-head Fab") (ME Brockson et al. (2014) Mol Microbiol 93: 1246–1258) and compared the result with that obtained using the corresponding intact IgG molecule. As a negative control, Fab fragments derived from a mouse monoclonal antibody against the 15-mer non-protective domain in the “tail” region of the IHF _NTHI β-subunit (“Fab against β-tail part”), which do not disrupt bacterial biofilms (LA Novotny et al. (2016) EBioMedicine 10: 33–44) or a non-specific mouse isotype IgG1 antibody (“isotype control Fab”) were used.

Representative images showed that each of the five different bacterial species tested formed biofilms of varying architecture within 24 h (Fig. 8A). Anti-β-tail Fab fragments or isotype control Fab fragments did not have a disruptive effect similar to that of intact IgG, from which these negative control fragments were derived. In contrast, incubation with anti-β-head Fab significantly disrupted biofilms formed by all bacterial species tested. Quantitative analysis of biofilm biomass showed that biofilm disruption by this concentration of anti-β-head Fab was comparable to that achieved by an equimolar concentration of the corresponding intact anti-β-head IgG (P ≤ 0.05; one-way ANOVA with multiple comparisons) (Fig. 8B). Thus, the minimal antigen-binding domain of the antibody molecule targeting the head portion is sufficient to bind and cleave the DNABII protein and subsequently disrupt the biofilm, and is as effective as the intact antibody from which the Fab fragments were derived.

Murine Fabs against the β-head removed NTHI from biofilms and eliminated biofilms during experimental OM

By identifying the biofilm-destroying function of anti-β-head Fab demonstrated in vitro, the applicant addressed the first question: in the context of experimental disease, is antibody-mediated cleavage of the DNABII protein via Fab domains alone sufficient to induce biofilm collapse and subsequent killing of released bacteria? To answer this question, the applicant utilized a well-established chinchilla model of OM based on the predominant causative agent of chronic and recurrent disease, NTHI. Experimental OM was first induced by direct infection of the middle ear. Four days later, NTHI biofilms filled the middle ear (Novotny LA et al (2011) Mucosal Immunol 2011; 4: 456–67). The resulting Fabs against the β-head region of the mouse monoclonal antibody, Fabs against the β-tail region, or isotype control Fabs were then injected into both middle ears with a second identical treatment 24 hours later (Fig. 9A). One day after receiving the second dose of Fabs, all animals were sacrificed to assess the relative immediate efficacy of treatment.

Applicant first quantified the amount of NTHI present in mucosal biofilms and/or adherent to the middle ear mucosa using plate counting. There was no difference in bacterial load in animals treated with anti-β-tail Fab fragments compared to isotype control Fabs (Fig. 9B). Conversely, 4 log fewer NTHI were detected in mucosal biofilms obtained from animals treated with anti-β-head Fab, and in one middle ear of six ears in this group, all NTHI were cleared (P≤0.01; one-way ANOVA with multiple comparisons) (Fig. 9B). Thus, delivery of anti-β-head Fab induced enhanced NTHI clearing in the middle ear of chinchillas.

Next, to determine whether any of the three treatments promptly dismantled established NTHI biofilms, one day after completion of Fab fragment therapy, images of each middle ear were obtained, randomized, and scored by six reviewers blinded to the experimental details. The reviewers used well-established scoring criteria, where a score of 0 corresponded to the absence of mucosal biofilm, and a score of 4+ indicated that ≥75% of the middle ear remained filled with mucosal biofilm. Accordingly, anti-β-tail Fab fragments induced a small but nonsignificant reduction in mucosal biofilm compared to delivery of isotype control Fab fragments. Importantly, ≥50% of the middle ears in the isotype control and anti-β-tail Fab groups remained filled with mucosal biofilm, with mean biomass scores of 3.4 and 2.7, respectively (Fig. 9C). Conversely, anti-β-head Fab induced a significant reduction in mucosal biofilm (P≤0.05; one-way ANOVA with multiple comparisons), and the majority (4 of 6, 67%) of the middle ears in this latter group were assigned a score ≤1.0, indicating that ≤25% of the middle ears contained any residual visible biofilm. Thus, delivery of anti-β-head Fab significantly reduced the bacterial load in the middle ear and induced efficient killing of established mucosal biofilms.

Representative images of the middle ear from each group are shown in Figure 9D, with the middle ear of a healthy chinchilla and an ear filled with NTHI biofilm shown for comparison. In naive animals, the middle ear mucosa and natural bony septa are visible throughout their length (score 0). In contrast, four days after NTHI challenge, neither the thin middle ear mucosa nor the bony septa are visible, as these anatomical features are now obscured by a large mucosal biofilm (score 4). In this study, after administration of anti-β-tail Fabs or control isotype Fabs, mucosal biofilms still filled 50–100% of the middle ear, with a representative assigned score of 4.0 or 2.4, respectively (Figure 9D). However, anti-β-head Fab largely destroyed these structures, as evidenced by the fact that both the middle ear mucosa and the bony septa were completely visible (assigned a score of 1.0). Also of note is that while extensive inflammation was observed in the middle ear mucosa and tympanic membrane of animals treated with anti-β-tail Fab (Fig. 9D, middle image, right side), only limited dilation of fine capillaries within the middle ear mucosa of animals treated with anti-β-head Fab (Fig. 9D, bottom image, right side) was observed in the middle ear mucosa of animals treated with anti-β-head Fab (Fig. 9D, bottom image, right side), despite these ears being filled with NTHI-formed biofilm just 2 days prior.

To begin to understand why the middle ears of animals receiving anti-β-tail Fab fragments were consistently visibly inflamed (Figure 9D, middle panel, right side), whereas ears receiving anti-β-head Fab fragments were not inflamed (Figure 9D, bottom panel, right side), the applicant determined a focused cytokine profile in middle ear fluids collected from animals at the time of sacrifice. Using bead cytometric analysis, significantly greater amounts of each of the six proinflammatory cytokines were detected in the middle ears of animals receiving anti-β-tail Fab fragments compared to animals receiving anti-β-head Fab fragments (P≤0.05) (Figure 10A). Conversely, significantly more anti-inflammatory cytokine IL-10 was detected in the middle ear fluid of animals treated with anti-β-head Fab (P≤0.05; one-way ANOVA with multiple comparisons) (Fig. 10B). Moreover, compared with animals treated with isotype control Fabs, significantly more proinflammatory cytokines IL-1β and TNF were detected in the middle ear fluid of animals treated with anti-β-head Fab (Fig. 10B), which likely contributed to the inflammation correspondingly observed in the mucous membranes in the latter group (see Fig. 9D). These data suggest that, in the context of active experimental OM, biofilm collapse and NTHI clearance induced by anti-β-head Fab also mediated the resolution of disease-associated inflammation. Taken together, and centered on the original question, the study results to date have shown that Fabs against the β-head region effectively disrupted bacterial biofilms in vivo, so the Fc portion of the antibody directed against DNA-II was not required to induce biofilm collapse. This finding is complemented by data from a model in which structural biofilm collapse with the release of resident bacteria results from an antibody-mediated equilibrium shift targeting the DNABII protein, which subsequently tips the balance in favor of the host, which now effectively eliminates newly released pathogens through interactions with multiple host immune effectors.

Rabbit IgG Fabs against domains in both IHF subunits rapidly disrupt biofilm in vivo.

To date, the applicant has investigated Fab fragments directed against the β-subunit, one of two heterologous subunits that comprise IHF _NTHI . While this approach has been effective, in a previous study, the applicant demonstrated that a mixture of murine monoclonal antibodies against the head domains within the α-subunit plus antibodies against the β-subunit of IHF _NTHI induced significantly greater biofilm disruption compared to antibodies against either subunit alone, including bacterial species possessing only the HU allele (LA Novotny et al. (2016) EBioMedicine 10: 33–44). Despite 74.7% amino acid sequence similarity, only 47.3% identity is observed within each 94-amino acid subunit of IHF _NTHI . Thus, a cocktail of antibodies directed against the α-subunit and β-subunit provides broader coverage of the entire diversity within IHF and its ortholog, HU. Using this information, the applicant constructed a chimeric peptide immunogen targeting the epitope to initially induce antibodies in rabbits against both protective domains simultaneously. A constructed “head chimeric peptide,” slightly larger (e.g., 20-mer) segments within the DNA-binding head domains of both the α-subunit and β-subunit of IHF _NTHI , were connected by a 4-mer linker peptide to provide flexibility between these two protective epitopes (LA Novotny et al. (2019) NPJ Vaccines 4: 43). As a negative control, a “tail chimeric peptide” immunogen was also constructed, which included 20-mer segments from the non-protective domains in the tail region of the α-subunit and β-subunit of IHF _NTHI and was connected by the same 4-residue linker. The applicant has previously shown that polyclonal rabbit IgG against this head-on chimeric peptide disrupts biofilms formed by several bacterial species in vitro and eliminates mucosal biofilms in the middle ears of chinchillas during experimental NTHI-induced OM (LA Novotny et al. (2019) NPJ Vaccines 4: 43).

To now combine the chimeric peptide design strategy with the identification of potential Fab-mediated therapeutic biofilm disruption, in this study, Applicant tested the ability of polyclonal rabbit Fabs against the head chimeric peptide to disrupt NTHI biofilms already present in the middle ears of chinchillas with experimental OM. Applicant also assessed the durability of any resulting biofilm disruption, i.e., would NTHI biofilms re-establish after cessation of antibody therapy? Accordingly, NTHI allowed the establishment of extensive biofilms in the chinchilla middle ear before Applicant treated with either Fabs derived from polyclonal rabbit IgG against 1) the head chimeric peptide; 2) the tail chimeric peptide; or 3) naive serum (Fig. 11A). One day after delivery of the second therapeutic dose, a subset of animals in each group was sacrificed to examine the immediate treatment effect. The remaining animals were observed for another seven days without further treatment to assess the sustainability of this therapeutic approach.

One day after receiving two doses of the anti-head chimeric peptide Fab, there was >2 log less NTHI in mucosal biofilms and/or mucosa-adherent biofilms in the groups receiving the indicated treatments compared with either of the two negative control groups (P≤0.05; one-way ANOVA with multiple comparisons) (Fig. 11B). After an additional week without further treatment, only a small reduction in NTHI load was observed within middle ear mucosal biofilms/mucosa-adherent biofilms in animals receiving the anti-tail chimeric peptide Fab or naive serum Fab. In contrast, at this latter time point, an additional 10-fold reduction in bacterial load was observed in animals receiving the anti-head chimeric peptide Fab (P≤0.05; one-way ANOVA with multiple comparisons). Notably, in 4 of 6 (67%) middle ears, these homogenized tissues were culture-negative for NTHI. These data suggest that NTHI released from the biofilm by Fab against the head chimeric peptide was subsequently removed by host immune effectors without any additional intervention or treatment.

The applicant then conducted a qualitative assessment of whether treatment with the designated anti-chimeric peptide Fabs was able to eradicate NTHI biofilms already present in the chinchilla middle ears. For this, reviewers blinded to the treatment administered were asked to rate the relative quantitative mucosal biofilm score remaining in the middle ears after treatment on a scale of 0 to 4+. Administration of anti-tail chimeric peptide Fabs was not effective. One day after treatment, animals treated with anti-tail chimeric peptide Fabs had slightly larger remaining biofilms than those in the middle ears of animals treated with naive serum Fabs, with mean quantitative biofilm scores of 3.2 and 2.6, respectively (Fig. 11C). In both of these negative control groups, >50% of the middle ears remained filled with mucosal biofilm. In contrast, in the group receiving the anti-head chimeric peptide Fab, all six middle ears scored ≤1.0, indicating that only minimal remaining mucosal biofilm was observed (P≤0.001; one-way ANOVA with multiple comparisons) with a mean biomass score of 0.7. Thus, the immediate effect of anti-head chimeric peptide Fab therapy was a significant reduction in bacterial load and significant disruption of established NTHI biofilms.

To assess the durability of the observed treatment effect, the middle ears were assessed in a subset of each group after an additional week without any additional treatment. In the group receiving naive rabbit serum Fab, mucosal biofilms actually increased, whereas they only decreased slightly in the group receiving anti-tail chimeric peptide Fab (e.g., >50% of middle ears remained filled with NTHI biofilm), with a mean biomass score of 3.6 and 2.6, respectively (Fig. 11C). Importantly, animals receiving anti-head chimeric peptide Fab continued to clear NTHI biofilms, as evidenced by a further significant reduction in mucosal biofilm (P≤0.01; one-way ANOVA with multiple comparisons) and a mean biomass score of 0.2. Notably, there was no visible evidence of biofilm in 4 of 6 (67%) middle ears. These qualitative assessments correlated well with the bacterial load data, as shown in Fig. 11B.

To demonstrate that reviewers blinded to the treatment administered performed the assessment on the remaining observable mucosal biofilms, examples of healthy middle ears and middle ears with a four-day NTHI biofilm, as well as representative images of middle ears from each group on day 13, are shown in Fig. 11D. As expected, one day after completion of treatment, animals receiving naive serum or anti-tail chimeric peptide Fab retained significant mucosal biofilms, as evidenced by the fact that these biofilms obscured the visibility of the middle ear mucosa and bony septa, with representative assigned scores of 2.8 or 3.2, respectively (Fig. 11D). These quantitative scores did not decrease significantly seven days after completion of treatment, and in fact, they increased markedly for the group receiving Fab from naive serum, with representative assigned scores of 4.0 and 2.8, respectively. Conversely, in the middle ears of animals treated with Fab against the head-mounted chimeric peptide, normal anatomical structures were fully visible one day after completion of treatment and remained so, with no evidence of biofilm regrowth, over the subsequent seven days. Representative assigned scores were 0.8 (day 6) and 0 (day 13) for this latter group. Taken together, these data provided further compelling support for the ability of Fab delivery against the head-mounted chimeric peptide to eradicate existing NTHI biofilms in the middle ear, and that this eradication likely continues after treatment.

Development and validation of a humanized version of a monoclonal antibody against the head chimeric peptide

With data supporting the use of Fabs against the head chimeric peptide for disrupting various bacterial biofilms in vitro, as well as for killing NTHI mucosal biofilms in vivo, the applicant wanted to move these new therapeutic agents into clinical trials. To this end, the applicant generated a panel of humanized monoclonal antibodies directed against the head chimeric peptide (herein referred to as "HuTipMAbs"), designed in the style of a murine monoclonal antibody against the head chimeric peptide. Here, the applicant presents data supporting the in vitro functional activity of one of the HuTipMabs to demonstrate the effectiveness of humanization. As an indicator of the degree of antibody humanization, the applicant defined the T20 value for the variable regions of the heavy and light chains separately (SH Gao et al. (2013) BMC Biotechnol 13: 55). These values were 82 for the heavy chain variable region and 97 for the light chain variable region, indicating a high degree of humanization for each antibody domain. According to surface plasmon resonance, HuTipMab had a _KD of 76 nM for the head chimeric peptide and a _{KD of} 10 nM for native IHF _NTHI . Thus, strong affinity for the target peptide and even greater affinity for the native protein, which is necessary for disease cure, were demonstrated. In vitro, the HuTipMab antibody significantly disrupted biofilms formed by NTHI, P. aeruginosa, or B. cenocepacia (P≤0.001; unpaired t-test) (Fig. 12A), and only 13–26% of biofilm biomass remained after 16-hour exposure to the selected concentration of the humanized Mab used, compared with biofilms incubated with an equivalent concentration of a humanized antibody against the tail chimeric peptide (called “HuTailMab”) (Fig. 12B). Thus, humanization of a murine monoclonal antibody specific for the head chimeric peptide resulted in a therapeutic agent with high affinity for its target protein and proven ability to disrupt biofilms formed by three human respiratory pathogens in vitro.

To further confirm that the humanized monoclonal antibody also functions in vivo, the applicant used a model of experimental NTHI-induced OM in chinchillas, in which biofilms were already present in the middle ear prior to treatment. Either HuTipMab, HuTailMab, or an equivalent volume of sterile saline (the diluent used with humanized antibodies) was injected into both middle ears, followed by a second identical treatment 24 hours later (Fig. 13A). As before, a subset of animals from each group was sacrificed one day after completion of therapy to examine immediate effects, and the second subset of animals was observed for an additional week without additional treatment to determine whether the treatment effect was maintained.

One day after treatment, HuTipMab induced a significant >3.5 log reduction in NTHI counts compared with HuTailMab or saline (P≤0.01; one-way ANOVA with multiple comparisons) (Fig. 13B). One week later, while either HuTailMab or saline induced only a small additional reduction in NTHI load, the load in HuTipMab-treated middle ears was reduced by an additional 7-fold. Notably, one week after HuTipMab treatment, homogenates from 4 of 6 middle ear mucosa (67%) were culture-negative (P≤0.05; one-way ANOVA with multiple comparisons). Thus, HuTipMab treatment caused rapid elimination of NTHI in middle ear mucosal biofilms.

To qualitatively assess how much mucosal biofilm biomass remained in the middle ear after treatment, the middle ear was again scored blindly on a scale of 0-4+. The middle ears of animals treated with saline or HuTailMab remained filled with mucosal biofilm that occupied >50% of the middle ear, and the mean biomass scores were 2.7 and 3.1, respectively (Fig. 13C and Fig. 13D; representative scores are indicated in the box in the lower right corner of each panel in Fig. 13D, first row). In contrast, the middle ears of animals treated with HuTipMab had minimal remaining mucosal biofilm (<25%), and the mean biomass score for the group was 1.0 (Fig. 13C and Fig. 13D). Seven days later, while mucosal biofilm actually increased in the groups receiving either saline (mean score 3.0) or HuTailMab (mean score 3.5), minimal biofilm was observed in the HuTipMab group (mean score 0.6) (Figs. 13C and 13D, representative scores are indicated in the box in the lower right corner of each panel in Fig. 13D, second row). Representative images for each group are shown in Fig. 13D and correlate well with the data presented in Fig. 13C.

In contrast to Fig. 9D, where inflammation was visible in the middle ears of animals injected with mouse monoclonal antibody fragments directed exclusively to the β-head portion of one of the IHF subunits, treatment with HuTipMab did not result in overt inflammation (see Fig. 13D). To explain this observation, applicant again performed bead cytometric analysis to determine the focused, but now slightly broadened, cytokine profile in the middle ear fluids collected from these animals at the time of sacrifice. One day after completion of antibody therapy, comparable amounts of six proinflammatory cytokines were detected in the middle ear fluids of animals treated with saline or HuTailMab, whereas significantly fewer were detected in the fluids of animals treated with HuTipMab (Fig. 14A). Additionally, significantly more of two anti-inflammatory cytokines were detected in the middle ear fluids obtained from animals treated with HuTipMab. This pattern was maintained and further enhanced in fluids collected on day 13 (7 days after completion of therapy) (Fig. 14B). Since proinflammatory cytokine concentrations were similar in saline- and HuTailMab-treated animals, without being bound by theory, applicants hypothesize that humanization of the mouse monoclonal antibody abolished the inflammation observed in animals in this latter group (see Fig. 13D). Collectively, these data demonstrated that humanization of the mouse monoclonal antibody directed against the head chimeric peptide does not reduce its efficacy either in vitro or in vivo. HuTipMab antibody induced rapid and robust clearance of NTHI-induced middle ear mucosal biofilms during experimental OM, thereby promoting disease resolution.

The biofilm disruption effects of HuTipMab and HuTailMab are summarized in the table below.

Table 9. Results of biofilm destruction in vivo

Antibody Heavy chain Light chain Biofilm destruction
in vivo % change in bacterial load after treatment % of occluded middle ears
after treatment Day 1 Day 8 Day 1 Day 8 HuTipMab SEQ ID NO: 1 SEQ ID NO: 7 ↓99 ↓99 <25% <25% HuTailMab SEQ ID NO: 4 SEQ ID NO: 10 ↑222 ↑796 50-<75% 50-<75

↓ indicates a decrease in bacterial load; while ↑ indicates an increase in bacterial load.

Furthermore, the applicant identified a limited cytokine profile in middle ear fluids collected from animals at the time of sacrifice. As shown in Fig. 14, significantly fewer proinflammatory cytokines were observed in the HuTipMab-treated middle ear, while significantly more anti-inflammatory cytokines were detected in the HuTipMab-treated middle ear. However, the HuTailMab-treated middle ear was comparable to the saline-treated middle ear in terms of the presence of proinflammatory cytokines. This suggests that humanization of the mouse mAb eliminated some of the rodent-specific inflammation observed in the Fab mAb study. This is a positive result for the use of HuTailMab as a diagnostic tool.

Preclinical efficacy of the head chimeric peptide after active transdermal immunization

Thus, while the applicant has developed an effective therapeutic agent targeting DNABII (e.g., delivery of a humanized monoclonal antibody that targets the immunoprotective head regions of both IHF subunits to disrupt biofilms/cure ongoing disease), preventing biofilm formation and disease induction by active immunization is also an important goal. To this end, the applicant utilized a unique experimental OM model of viral-bacterial co-infection of chinchillas, in which a prior adenovirus infection predisposes the middle ear to invasion by NTHI bacteria colonizing the nasopharynx, which now ascend the virus-infected eustachian tube (K. Suzuki et al. (1994) Infect Immun 62: 1710-1718). This superinfection model is designed to mimic the event "My child has a cold, and a week later he has an ear infection." Thus, animals were first actively immunized with the head or tail chimeric peptide to induce the appropriate immune response (L. A. Novotny et al. (2015) Clin Vaccine Immunol 22: 867–874; L. A. Novotny et al. (2013) Vaccine 31: 3417–3426; L. A. Novotny et al. (2011) Mucosal Immunol 4: 456–467; L. A. Novotny et al. (2017) Clin Vaccine Immunol 24). Chinchillas were then challenged first with adenovirus and 4 days later with NTHI, after which the relative development of ascending OM was observed.

Three groups of naive chinchillas were immunized by rubbing vaccine formulations into the skin just behind both ears (i.e., the retroauricular region) and repeated one week later (Fig. 15A). The following formulations were used: a head chimeric peptide adjuvanted with LT(R192G/L211A) (dmLT), a heat-labile enterotoxin from double mutant E. coli (J.D. Clements et al. (2018) mSphere 2018 Jul-Aug; 3(4): e00215-18.); a tail chimeric peptide plus dmLT, or dmLT alone. Two days after the second immunization, all animals were inoculated intranasally with adenovirus and one week later with intranasal NTHI. Nasopharyngeal lavage was performed one day after NTHI challenge to confirm that all animals were equally colonized (Fig. 15B). This result ensured that each animal had the potential to develop experimental OM, with the only distinguishing factor being the induced immunity resulting from the immunogen administered.

The primary measure of vaccine efficacy was videootoscopy, in which each tympanic membrane (TM) was examined without knowledge of treatment to determine whether overt inflammation and middle ear fluid, signs of OM, were visible (L. A. Novotny et al. (2006) Vaccine 24: 4804–4811). Within 8 days after NTHI challenge, virus-infected chinchillas showed evidence that NTHI had ascended the weakened eustachian tube and induced OM in 2 of 16 middle ears (13%) in the adjuvant-only group, with a peak incidence of 100% on day 13 (Fig. 15C). Experimental OM began to decline after day 13 but persisted in 8 of 16 middle ears (50%) on day 20. As expected, immunization with the tail chimeric peptide did not provide an advantage. Evidence of OM was present in two middle ears (13%) at four days post-NTHI challenge, peaking at 100% on days 12 and 13 before beginning to decline, but 8 of 16 middle ears (50%) retained evidence of OM at the end of the study. In contrast, there was a significant delay in the onset of OM in the head chimeric peptide immunized group (P≤0.0001; Mantel-Cox test) until day 10. The peak OM frequency on days 10–12 was observed in only 4 of 18 middle ears (25%) in this latter group, with complete resolution by day 14, which was significantly earlier than in controls (P≤0.0001; Mantel-Cox test). Subsequently, over the 20-day observation period, the proportion of animals with signs of experimental OM was significantly lower in those immunized with the head chimeric peptide (4 ears; 2 animals; P≤0.0001; Mantel-Cox test) compared to the other two groups (16 ears; 8 animals).

The images of the chinchilla TM shown in Fig. 15D represent the results of otoscopy without knowledge of the treatment on day 11. While the TM of the healthy chinchilla is gray, the TM of animals immunized with dmLT or the tail chimeric peptide were slightly swollen, erythematous, and yellow middle ear fluid was visible behind the TM. In contrast, the TM of most animals in the group immunized with the head chimeric peptide (12 of 16; 75%) showed minimal signs of inflammation, if any. Overall, the vaccine efficacy provided by immunization with the head chimeric peptide was 85% compared to any negative control group. Taken together, these data support the development and use of the head chimeric peptide as an effective candidate antigen for a vaccine to prevent the development of experimental OM caused by NTHI and complement the applicant's development of a therapeutic humanized monoclonal antibody.

Discussion of the experiment

Biofilms are involved in most chronic and recurrent bacterial diseases of the respiratory tract (L. O. Bakaletz et al. (2012) Paediatr Respir 13: 154–159; J. H. Fastenberg et al. (2016) World J Otorhinolaryngol Head Neck Surg 2: 219–229; J. W. Costerton (2001) Trends Microbiol 9: 50–52), oral cavity (W. H. Bowen et al. (2018) Trends Microbiol 26: 229–242; P. E. Petersen et al. (2005) J Periodontol 76: 2187–2193), gastrointestinal tract (E. C. von Rosenvinge et al. (2013) Pathog Dis 67: 25–38; S. Macfarlane et al. (2007) J Appl Microbiol 102: 1187–1196) and the genitourinary tract (A.L. Flores-Mireles et al. (2016) J Urol 196: 416–421; P. Tenke et al. (2012) World J Urol 30: 51–57). The formation and persistence of these common polymicrobial communities significantly contributes to the pathogenesis of these diseases, largely due to their inherent resistance to clearance by host immune effectors and antibiotics. Accordingly, recognition of the role of biofilms in disease pathogenesis and persistence requires new approaches to either prevent their formation or eliminate those already present.

Thus, many laboratories have developed a variety of approaches to reduce the negative impacts of biofilm, which have been the subject of many excellent reviews (Verderosa AD et al. (2019) Front Chem 2019; 7: 824; Koo H et al. (2017) Nat Rev Microbiol 2017; 15: 740–55; Fleming D et al. (2017) Microorganisms 2017; 5: 15; Worthington RJ (2012) Org Biomol Chem 2012; 10: 7457–74; Reza A et al. (2019) Antibiotics (Basel) 2019; 8: 229). A wide area of research focuses on biofilm killing using agents that induce the proliferation of biofilm-resident bacteria, i.e. on the processing of biofilms by enzymes, molecules that interfere with processes such as bacterial quorum sensing, signaling through cyclic di-GMP, or the delivery of small molecule inhibitors or analogs (Verderosa AD et al. (2019) Front Chem 2019; 7: 824; Koo H et al. (2017) Nat Rev Microbiol 2017; 15: 740–55; Fleming D et al. (2017) Microorganisms 2017; 5: 15; Worthington RJ (2012) Org Biomol Chem 2012; 10: 7457–74; Reza A et al. (2019) Antibiotics (Basel) 2019; 8: 229). Alternatively, preventing biofilm formation is an approach that may also limit disease-induced inflammation, which is often more harmful to the host than the presence of the biofilm itself (Vestby LK et al. (2020) Antibiotics (Basel) 2020; 9: 59). Modification of implantable devices or incorporation of inhibitors into biomaterials, use of antimicrobial peptides or blockade of adhesion proteins expressed by microorganisms have been shown to limit biofilm formation (Bazaka K et al. (2012) Appl Microbiol Biotechnol 2012; 95: 299–311; Qvortrup K et al (2019) Front Chem 2019; 7: 742; Rabin N et al (2015) Future Med Chem 2015; 7: 647–71; Chung PY et al (2017) J Microbiol Immunol Infect 2017; 50: 405–10).

Over the past 10+ years, since the presenter identified the eDNA-DNABII lattice in biofilms formed by NTHI in the middle ear of chinchillas (S.D. Goodman et al. (2011) Mucosal Immunol 4: 625–637), this observation has been extended to demonstrate the presence of this bacteria-permissive but host-restrictive lattice in biofilms formed by many different bacterial species, including the high-priority ESKAPE pathogens (A. Abu Khweek et al. (2013) Front Cell Infect Microbiol 3: 18; A. Devaraj et al. (2015) Mol Microbiol 96: 1119–1135; M.O. Freire et al. (2017) Mol Oral Microbiol 32: 74–88; J.E. Gustave et al. (2013) J Cyst Fibros 12: 384-389; L.A. Novotny et al. (2013) PLoS One 8: e67629; L.A. Novotny et al. (2016) EBioMedicine 10: 33-44; C.J. Rocco et al. (2018) J Bacteriol 200; K.M. Rood et al. (2018) Sci Rep 8: 8756). Taken together, this observation, initially made in vitro and then extended to testing in multiple preclinical models (Goodman SD et al (2011) Mucosal Immunol 2011; 4: 625–37; Brockson ME et al (2014) Mol Microbiol 2014; 93: 1246–58; Novotny LA et al (2019) NPJ Vaccines 2019; 4: 43; Freire MO et al (2017) Mol Oral Microbiol 2017; 32: 74–88; and Novotny LA et al (2016) EBioMedicine 2016; 10: 33–44), supported the development of a novel DNABII-targeted approach for biofilm disease intervention that would target this seemingly species-independent ‘Achilles heel’.

Thus, a two-pronged approach to combating bacterial biofilms was developed and presented, both targeting the eDNA+DNABII scaffold of the bacterial biofilm. The first approach is a preventative active immunization strategy, in which the head-mounted chimeric peptide is used as an immunogen to induce antibody production that blocks biofilm formation via NTHI in the middle ear and, thus, the development of experimental OM. The applicant developed a candidate vaccine antigen based on the epitope-specific DNABII chimeric protein to induce antibody production that would prevent biofilm formation following active immunization. The second approach is a therapeutic strategy in which IgG or Fab antibody fragments directed against the head-mounted chimeric peptide immunogen (and ultimately a humanized monoclonal antibody) are delivered directly to the middle ear to disrupt established NTHI biofilms and mediate cure of experimental OM. The applicant has humanized a murine monoclonal antibody against this immunogen for use either as intact antibodies or as Fab fragments as a broadly effective therapeutic agent that releases bacteria from biofilm, which can then be killed by host immune effectors and/or antibiotics, which can now be used at a significantly reduced dose (M. Shigeta et al. (1997) Chemotherapy 43: 137-141; D.M. Lewis (2005) J Theor Biol 234: 565-591; S. Singh et al. (2017) Open Microbiol J 11: 53-62). Here, the applicant presented promising preclinical data from four studies using two different OM models that consistently showed how effective these approaches can be in terms of biofilm disruption and disease prevention and/or markedly and significantly reducing existing mucosal biofilms with rapid disease resolution.

The applicant used two preclinical models of a common childhood disease, the chronic course and recurrence of which is caused by biofilms in the middle ear, to evaluate the prophylactic and therapeutic efficacy of the applicant's DNABII-targeted strategies. These models closely mimic the natural history of OM observed in children (G.S. Giebink (1999) Microb Drug Resists 5: 57–72; L.O. Bakaletz (2009) Expert Rev Vaccines 8: 1063–1082; J.T. Poolman et al. (2000) Vaccine 19 Suppl 1: S108–115) and accurately predicted clinical trial outcomes (L.A. Novotny et al. (2006) Vaccine 24: 4804–4811; R. Prymula et al. (2006) Lancet 367: 740–748). Importantly for the studies presented herein, the chinchilla is a well-established host in which to study the formation, persistence, and invasion of mono- and polymicrobial biofilms (L.O. Bakaletz (2012) Paediatr Respir Rev 13: 154-159; G.D. Ehrlich et al. (2002) JAMA 287: 1710-1715; M.A. Apicella (2009) J Infect Dis 199: 774-775).

The applicant provided answers to several previously unanswered questions. The applicant found that Fab fragments are as effective as the intact antibody at disrupting biofilms in vitro and in vivo and thus may serve as a therapeutic option when repeated dosing may be required (due to concerns about the induction of an antibody response). The applicant also determined that the Fc portion of the DNA-BII-targeted antibody is not required for biofilm disruption in vitro or for biofilm eradication in vivo. This result suggests that the release of bacteria from the protective biofilm will be sufficient to facilitate clearance and disease resolution by host innate immune effectors and/or antibiotics. The applicant found that the inclusion of protective domains from both heterogeneous subunits of the IHF _NTHI does indeed result in increased efficacy. Furthermore, active immunization with the head chimeric peptide induced the formation of antibodies that prevented the development of experimental OM and facilitated rapid disease recovery. Humanization of the monoclonal antibody to the head chimeric peptide allowed the creation of a promising therapeutic agent with nanomolar affinity for both the immunogen and the native protein.

Taken together, these data contributed to the understanding of the eDNA+DNABII-targeted strategy and significantly initiated the initiation of both prophylactic and therapeutic clinical trials of both the head chimeric peptide immunogen and the humanized monoclonal antibody against it, respectively. Given the species-independent nature of the applicant's DNABII-targeted approach and its non-theoretical nature, this offers a therapeutic product for improved clinical application in a variety of diseases in which the pathogenesis, persistent chronicity, and/or cyclic recurrence are mediated by non-degradable biofilm.

Experiment #5

A number of oral bacteria (e.g., Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis) are involved in the pathogenesis of inflammatory diseases such as periodontitis and peri-implantitis, which destroy alveolar bone and gingiva. Research into the pathogenesis of these bacteria is hampered by the lack of effective animal models. One of the challenges in studying the pathogenicity of specific bacteria is the difficulty of biofilm formation when introducing exogenous bacteria into the oral cavity of animals. Although animal models of periodontitis have been developed, cultured bacteria are rarely isolated from the oral cavity of inoculated animals. The development of an effective animal model that can assess the pathogenicity of specific bacteria will greatly aid in elucidating the mechanisms of their pathogenesis.

The applicants created a rat model of peri-implantitis in which bacterial biofilms (e.g., A. actinomycetemcomitans, P. gingivalis) are grown on the heads of titanium screw implants (Freire, M.O., (2011) J. Periodontology 82(5):778–89 and Freire, M.O., (2017) Molecular Oral Microbiology 32(1):74–88). The biofilm-laden screws are then surgically implanted into the aveolar bone of the rat maxilla and observed over time. Microcomputed tomography can be used to measure bone loss around the screws, and DNA extracted from the screws, as well as tissue and bone surrounding the screws, can be used for quantitative PCR or microbiome analysis to identify the species present. This established animal model demonstrated the presence of bone loss around the screw implant and the detection of inoculated bacterial pathogens at least 2 weeks after implantation.

Experiment #6

This experiment presents a mouse model for preclinical testing of interfering agents for the treatment of Lyme disease. See Dresser et al. Pathogens 5(12)e1000680, Epub 2009 Dec. 4. Lyme disease is the most common tick-borne disease in the United States. The number of reported cases more than doubled between 1992 and 2006, with approximately 29,000 new cases confirmed in 2008. It is estimated that the actual number of Lyme disease cases may be 6-12 times higher than those reported in endemic areas. By definition, these endemic areas are expanding as populations continue to move from cities to suburbs and rural areas, and white-tailed deer (carriers of Ixodes ticks) increasingly roam these areas. Lyme disease is caused by the microorganism Borrelia burgdorferi, a spirochete. B. burgdorferi is transmitted through the bite of an Ixodid tick and subsequently spreads through the bloodstream to other tissues and organs.

In this animal model, C3H/HeN mice were inoculated with spirochetes via dorsal, subcutaneous, and intraperitoneal injections, or via intravenous injection. Blood samples and biopsies were collected approximately 7 days post-infection to assess microbial load and evaluate tissue and organ pathology. The methods and compositions disclosed herein are intended to develop both therapeutic and prophylactic strategies for reducing and/or eliminating B. burgdorferi biofilms that form after infection and are believed to contribute to both the pathogenesis and chronicity of the disease.

Experiment #7

This experiment represents a porcine model for preclinical testing of interfering agents for the treatment of cystic fibrosis. See Stoltz et al. (2010) Science Translational Medicine 2(29):29–31. Cystic fibrosis is an autosomal recessive disease due to mutations in the gene that encodes the CF transmembrane conductance regulator anion channel (called CFTR). In this model, pigs that were specifically bred to carry a defect in genes called "CFTR" and called CF pigs spontaneously developed the characteristic features of CF lung disease, which included lower respiratory tract infection with several bacterial species. Pigs can be immunized with interfering agents to 1) immunize CF pigs with a polypeptide or other immunogenic agent, thereby inducing the production of antibodies that will destroy bacterial biofilms in the lungs, 2) deliver antibodies, or their fragments or derivatives, to the lungs of these animals by nebulization to assess the reduction in signs of the disease and associated pathologies.

Experiment #8

The applicants also presented a preclinical model of tuberculosis (TB). See Ordway et al. (2010) Anti. Agents and Chemotherapy 54:1820. Mycobacterium tuberculosis is the cause of a growing global epidemic. According to current data, approximately 8 million new cases of TB and approximately 2.7 million TB-related deaths are reported annually. In addition to the microbe's role as a coinfection in people with HIV (of the estimated 45 million people infected with HIV, ~30% are also infected with M. tuberculosis), a particular concern is that isolates have become highly resistant to multiple drugs, and no new anti-TB drug has been developed for more than a quarter century. In this animal model of SPF, guinea pigs were housed in a barrier colony and infected with an aerosol spray to deliver ~20 CFU of M. tuberculosis Erdman K01 strain bacilli to their lungs. Animals were sacrificed, bacterial loads were determined, and tissues were collected for histopathological evaluation at 25, 50, 75, 100, 125, and 150 days postinfection. Unlike mice, which do not develop classic signs of tuberculosis, guinea pigs infected in this manner develop well-organized granulomas with central necrosis, a hallmark of the human disease. Subsequently, as in humans, guinea pigs develop severe pyogranulomatous and necrotizing lymphadenitis of the draining lymph nodes as part of the primary lesion complex. Use of this model will provide preclinical screening to validate and identify therapeutic as well as prophylactic strategies to reduce and/or eliminate the resulting M. tuberculosis biofilms that have been observed to form in the lungs of these animals following infection and are thought to contribute to both the pathogenesis and chronicity of the disease.

Experiment #9

Several animal models of catheter/implanted device biofilm infection are known. See Otto (2009) Nature Reviews Microbiology 7:555. Although typically considered normal skin flora, Staphylococcus epidermidis has become what many consider a key opportunistic pathogen, ranking first among the causative agents of hospital-acquired infections. Most notably, this bacterium is responsible for the majority of infections that develop on implanted medical devices, which are contaminated with this common skin colonizer during device insertion. Although typically not life-threatening, the difficulties associated with treating these biofilm infections, coupled with their frequency, make them a significant public health burden. The ongoing costs associated with treating vascular catheter-associated bloodstream infections caused by S. epidermidis alone amount to $2 billion annually in the United States. In addition to C. epidermidis, E. faecalis and S. aureus are also contaminants found on implanted medical devices. Several animal models of catheter-associated S. epidermidis infections exist, including rabbits, mice, guinea pigs, and rats, all of which are used to study the molecular mechanisms of pathogenesis and are amenable to research in prevention and/or therapy. Rat jugular vein catheters have been used to evaluate treatments that interfere with E. faecalis, S. aureus, and S. epidermidis biofilm formation. Biofilm reduction is often measured in three ways: (i) sonicating the catheter and counting the CFU, (ii) cutting off pieces of the catheter or simply placing them on a plate and scoring them, or (iii) staining the biofilm with crystal violet or another dye, eluting, and measuring the OD as an indicator of CFU.

Experiment #10

The methods described in this application can be used to induce immune responses in humans and animals. Immunogenic compositions can be administered to humans and animals in the presence of adjuvants, such as, but not limited to, aluminum salts and liposomes. Those skilled in the art will appreciate that any number of pharmaceutically acceptable adjuvants can also be used. Immunogenic compositions can be administered to humans or animals intramuscularly, subcutaneously, intranasally, or by any other suitable route. Immunogenic compositions can be prepared in a manner appropriate to the selected route of administration. Immunogenic compositions can be in the form of polypeptides, nucleic acids, or a combination thereof, and can include full-length or partial antigens. Additionally or alternatively, immunogenic compositions can be in the form of APCs activated by a specific antigen, or APCs transfected with one or more polynucleotides encoding a specific antigen. Administration can include a single dose of the immunogenic composition or an initial administration followed by one or more booster doses. Booster doses can be administered one day, two days, three days, one week, two weeks, three weeks, one, two, three, six, or twelve months after the initial dose, or at any other time. The booster dose can be administered after the subject's antibody titer has been assessed.

Experiment #11

The methods described in this application can be used to generate passive immunity in a non-immune subject. Passive immunity against a given antigen can be achieved by transferring antibodies or antigen-binding fragments that specifically recognize or bind to a particular antigen. Antibody donors and recipients can be humans or non-human subjects. Additionally or alternatively, the antibody composition can comprise an isolated or recombinant polynucleotide encoding an antibody or antigen-binding fragment that specifically recognizes or binds to a particular antigen.

Passive immunity can be provided in cases where the administration of immunogenic compositions poses a risk to the recipient subject, the recipient subject is immunocompromised, or the recipient subject requires immediate immunity. Immunogenic compositions can be prepared in a manner appropriate to the selected route of administration. The compositions can contain full-length antibodies, antigen-binding fragments, polyclonal antibodies, monoclonal antibodies, antibodies produced in vivo, antibodies produced in vitro, purified or partially purified antibodies, or whole serum. Administration can include a single dose of the antibody composition or an initial administration followed by one or more booster doses. Booster doses can be administered one day, two days, three days, one week, two weeks, three weeks, one, two, three, six, or twelve months later, or at any other time after the initial dose. A booster dose can be administered after assessing the antibody titer of the subject.

Equivalents

It should be understood that, while the invention has been described in connection with the embodiments described above, the preceding description and examples are intended to illustrate and not limit the scope of the invention. Other aspects, advantages, and modifications within the scope of the invention will be apparent to those skilled in the art to which the invention pertains.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. All nucleotide sequences presented herein are presented in the 5' to 3' direction.

The embodiments illustratively described in this application may be properly practiced in the absence of any element or elements, limitation or limitations not specifically disclosed herein. Thus, for example, the terms "comprising," "including," "encompassing," etc. should be understood broadly and without limitation. Furthermore, the terms and expressions used in this description are used as terms of description and not as limitations, and there is no intention in the use of such terms and expressions to exclude any equivalents, disclosed and described functions or portions thereof, but it is recognized that various modifications are possible within the scope of the invention.

It should therefore be understood that, although the present invention has been specifically disclosed through certain embodiments and additional features, modifications, improvements, and variations of the embodiments disclosed herein may be made by those skilled in the art, and that such modifications, improvements, and variations are considered to be within the scope of this invention. The materials, methods, and examples presented in this description are representative of specific embodiments, are illustrative, and are not intended to limit the scope of the invention.

The scope of the invention is presented broadly and generally in this description. Each narrower embodiment falling within the general disclosure of the invention also forms part of the invention. This includes a general description with a condition or negative limitation excluding any object from the embodiment, regardless of whether the excluded object is specifically stated in this description or not.

Furthermore, where features or aspects of the invention are described in terms of the Markush formula, those skilled in the art will appreciate that embodiments of the invention may also thereby be described in terms of any individual compound or subgroup of compounds of the Markush formula.

All publications, patent applications, patents, and other references mentioned in this specification are expressly incorporated by reference in their entirety to the same extent as if each were individually incorporated by reference. In case of conflict, this specification, including definitions, will control.

NON-LIMITING EMBODIMENTS of the invention

Embodiment 1. An antibody or fragment thereof, comprising or alternatively consisting essentially of or additionally consisting of:

(i) an immunoglobulin heavy chain (HC) variable domain sequence comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group of amino acids (aa) 25-144 of SEQ ID NO: 13, 24 or 26 or the equivalent of each; and/or

(ii) an immunoglobulin light chain (LC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-132 of SEQ ID NO: 14 or 25, aa 21-126 of SEQ ID NO: 27, or the equivalent of each.

Embodiment 2. An antibody or fragment thereof according to embodiment 1, characterized in that the antibody comprises or consists essentially of or additionally consists of:

(i) an immunoglobulin heavy chain (HC) variable domain sequence comprising or alternatively consisting essentially of or additionally consisting of aa 25-144 of SEQ ID NO: 24 or an equivalent thereof; and/or

(ii) an immunoglobulin light chain (LC) variable domain sequence comprising or alternatively consisting essentially of or additionally consisting of the sequence aa 22-132 of SEQ ID NO: 25 or an equivalent thereof.

Embodiment 3. An antibody or fragment thereof according to embodiment 1, characterized in that the antibody comprises or consists essentially of or additionally consists of:

(i) an immunoglobulin heavy chain (HC) variable domain sequence comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or the equivalent of each; and/or

(ii) an immunoglobulin light chain (LC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or the equivalent of each of them.

Embodiment 4. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 5. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 6. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 7. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 8. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 9. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 10. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-132 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 11. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-132 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 12. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-132 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 13. The antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or consists essentially of or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, consists essentially of or additionally consists of an amino acid sequence of aa 21-126 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 14. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence of aa 21-126 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 15. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of or additionally consists of an amino acid sequence of aa 21-126 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 16. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 17. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 18. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 19. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 20. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 21. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 22. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 23. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 24. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 25. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 26. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 27. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 28. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, consists essentially of, or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, consists essentially of, or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 29. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 30. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, consists essentially of, or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, consists essentially of, or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 31. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 32. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 5-144 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 33. An antibody or fragment thereof according to embodiment 1, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 34. An antibody or fragment thereof, comprising or alternatively consisting essentially of or additionally consisting of:

(i) an immunoglobulin heavy chain (HC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group aa 25-144 of SEQ ID NO: 13, 24 or 26; and/or

(ii) an immunoglobulin light chain (LC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group aa 21-132 of SEQ ID NO: 14 or 25, aa 21-126 of SEQ ID NO: 27.

Embodiment 35. An antibody or fragment thereof according to embodiment 34, characterized in that said antibody comprises or consists essentially of or additionally consists of:

(i) an immunoglobulin heavy chain (HC) variable domain sequence comprising or alternatively consisting essentially of or additionally consisting of the sequence aa 25-144 of SEQ ID NO: 24; and/or

(ii) an immunoglobulin light chain (LC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of the sequence aa 21-132 of SEQ ID NO: 25.

Embodiment 36. An antibody or fragment thereof according to embodiment 34, characterized in that said antibody comprises or consists essentially of or additionally consists of:

(i) an immunoglobulin heavy chain (HC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26; and/or

(ii) an immunoglobulin light chain (LC) variable domain sequence comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 37. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or consists essentially of the amino acid sequence aa 25-144 of SEQ ID NO: 1, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or consists essentially of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 38. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 39. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or consists essentially of the amino acid sequence aa 25-144 of SEQ ID NO: 3, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or consists essentially of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 40. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 41. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or consists essentially of the amino acid sequence aa 25-144 of SEQ ID NO: 5, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or consists essentially of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 42. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-132 of SEQ ID NO: 7-9, 14 or 25, aa 21-126 of SEQ ID NO: 10-12 or 27.

Embodiment 43. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-132 of SEQ ID NO: 7.

Embodiment 44. An antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-132 of SEQ ID NO: 8.

Embodiment 45. An antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-132 of SEQ ID NO: 9.

Embodiment 46. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of, or additionally consists of, an amino acid sequence of aa 21-126 of SEQ ID NO: 10.

Embodiment 47. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence of aa 21-126 of SEQ ID NO: 11.

Embodiment 48. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises, essentially consists of, or additionally consists of an amino acid sequence selected from the group of aa 25-144 of SEQ ID NO: 1-6, 13, 24 or 26, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of, or additionally consists of, an amino acid sequence of aa 21-126 of SEQ ID NO: 12.

Embodiment 49. An antibody or fragment thereof according to embodiment 34, wherein the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or consists essentially of the amino acid sequence aa 25-144 of SEQ ID NO: 1, and/or wherein the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or consists essentially of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 7.

Embodiment 50. An antibody or fragment thereof according to embodiment 34, wherein the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 25-144 of SEQ ID NO: 1, and/or wherein the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 8.

Embodiment 51. An antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or mainly consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 1, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or mainly consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 9.

Embodiment 52. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 7.

Embodiment 53. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 8.

Embodiment 54. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 2, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 9.

Embodiment 55. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 7.

Embodiment 56. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 8.

Embodiment 57. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 25-144 of SEQ ID NO: 3, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-132 of SEQ ID NO: 9.

Embodiment 58. An antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 10.

Embodiment 59. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 11.

Embodiment 60. An antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 4, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 12.

Embodiment 61. An antibody or fragment thereof according to embodiment 34, wherein the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5, and/or wherein the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 21-126 of SEQ ID NO: 10.

Embodiment 62. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 11.

Embodiment 63. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 5, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 12.

Embodiment 64. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of the amino acid sequence aa 21-126 of SEQ ID NO: 10.

Embodiment 65. An antibody or fragment thereof according to embodiment 34, wherein the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6, and/or wherein the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 11.

Embodiment 66. The antibody or fragment thereof according to embodiment 34, characterized in that the sequence of the variable domain of the heavy chain (HC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-144 of SEQ ID NO: 6, and/or characterized in that the sequence of the variable domain of the light chain (LC) of the immunoglobulin comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-126 of SEQ ID NO: 12.

Embodiment 67. An antibody or fragment thereof, comprising or alternatively consisting essentially of or additionally consisting of:

(i) a heavy chain (HC) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group aa 25-473 of SEQ ID NO: 13, 24 or 26, or an equivalent of each; and/or

(ii) a light chain (LC) comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group: aa 21-239 of SEQ ID NO: 14 or 25, aa 21-233 of SEQ ID NO: 27, or an equivalent of each of them.

Embodiment 68. An antibody or fragment thereof according to embodiment 67, characterized in that said antibody comprises, consists essentially of, or additionally consists of:

(i) a heavy chain (HC) comprising or alternatively consisting essentially of or additionally consisting of the sequence aa 25-473 of SEQ ID NO: 24 or an equivalent thereof; and/or

(ii) a light chain (LC) comprising or alternatively essentially consisting of or additionally consisting of the sequence aa 21-239 of SEQ ID NO: 25 or an equivalent thereof.

Embodiment 69. An antibody or fragment thereof according to embodiment 67, characterized in that said antibody comprises, consists essentially of, or additionally consists of:

(i) a heavy chain (HC) comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26, or an equivalent of each; and/or

(ii) a light chain (LC) comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from the group of aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 70. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 71. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 72. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 73. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 74. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 75. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from the group aa 21-239 of SEQ ID NO: 7-9, 14 or 25, aa 21-233 of SEQ ID NO: 10-12 or 27, or an equivalent of each of them.

Embodiment 76. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of an amino acid sequence selected from the group of aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of aa 21-239 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 77. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of an amino acid sequence selected from the group of aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of aa 21-239 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 78. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of an amino acid sequence selected from the group of aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of aa 21-239 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 79. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of an amino acid sequence selected from the group aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence aa 21-233 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 80. An antibody or fragment thereof according to embodiment 67, wherein the heavy chain (HC) comprises or essentially consists of an amino acid sequence selected from the group aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence aa 21-233 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 81. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of an amino acid sequence selected from the group aa 25-473 of SEQ ID NO: 1-6, 13, 24 or 26 or an equivalent of each of them, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence aa 21-233 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 82. An antibody or fragment thereof according to embodiment 67, wherein the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 1 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 83. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 84. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 1 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 85. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 86. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 2 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 87. An antibody or fragment thereof according to embodiment 67, wherein the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 2 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 88. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 89. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 3 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 90. An antibody or fragment thereof according to embodiment 67, wherein the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 3 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-239 of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 91. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 92. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 93. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 4 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 94. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of the amino acid sequence aa 25-473 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 95. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-473 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 96. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-473 of SEQ ID NO: 5 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 97. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-473 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 98. The antibody or fragment thereof according to embodiment 67, characterized in that the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-473 of SEQ ID NO: 6 or an equivalent thereof, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 99. An antibody or fragment thereof according to embodiment 67, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 25-473 of SEQ ID NO: 6 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence aa 21-233 of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 100. An antibody or fragment thereof, comprising or alternatively essentially consisting of or additionally consisting of:

(i) a heavy chain (HC) comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from SEQ ID NO: 13, 24 or 26, or an equivalent of each; and/or

(ii) a light chain (LC) comprising or alternatively essentially consisting of or additionally consisting of a sequence selected from SEQ ID NO: 14, 25 or 27, or an equivalent of each.

Embodiment 101. An antibody or fragment thereof according to embodiment 100, characterized in that said antibody comprises, consists essentially of, or additionally consists of:

(i) a heavy chain (HC) comprising or alternatively consisting essentially of or additionally consisting of the sequence of SEQ ID NO: 24 or an equivalent thereof; and/or

(ii) a light chain (LC) comprising or alternatively consisting essentially of or additionally consisting of the sequence of SEQ ID NO: 25 or an equivalent thereof.

Embodiment 102. An antibody or fragment thereof according to embodiment 100, characterized in that said antibody comprises, consists essentially of, or additionally consists of:

(i) a heavy chain (HC) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from SEQ ID NO: 1-6, 13, 24 or 26, or an equivalent of each; and/or

(ii) a light chain (LC) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each.

Embodiment 103. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each of them.

Embodiment 104. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each of them.

Embodiment 105. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each of them.

Embodiment 106. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each of them.

Embodiment 107. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each of them.

Embodiment 108. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each of them.

Embodiment 109. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each of them, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 110. An antibody or fragment thereof according to embodiment 100, characterized in that the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each of them, and/or characterized in that the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 111. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each of them, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 112. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each of them, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 113. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each of them, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 114. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of an amino acid sequence selected from SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each of them, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of an amino acid sequence of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 115. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 116. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 117. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 118. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 119. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 120. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 121. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 7 or an equivalent thereof.

Embodiment 122. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 8 or an equivalent thereof.

Embodiment 123. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 9 or an equivalent thereof.

Embodiment 124. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 125. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 126. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 127. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 128. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 129. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 130. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 10 or an equivalent thereof.

Embodiment 131. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 11 or an equivalent thereof.

Embodiment 132. An antibody or fragment thereof according to embodiment 100, wherein the heavy chain (HC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and/or wherein the light chain (LC) comprises or essentially consists of or additionally consists of the amino acid sequence of SEQ ID NO: 12 or an equivalent thereof.

Embodiment 133. An antibody or fragment thereof, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or further consisting of a sequence selected from the group: GFTFXXY (amino acids (aa) 50-56 of SEQ ID NO: 13), GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24), or GFTFSRY (aa 50-56 of SEQ ID NO: 4 or 5 or 6 or 26), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 1-6;

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or further consisting of a sequence selected from the group: XSXXXX (amino acids (aa) 76-81 of SEQ ID NO: 13), GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24), or SSGGSY (aa 76-81 of SEQ ID NO: 4 or 5 or 6 or 26), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 1-6;

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: XXXXXXXYXXFDX (amino acids (aa) 121-133 of SEQ ID NO: 13), VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24), or ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 1-6;

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or further consisting of a sequence selected from the group: QXXXXXXXXXX (aa 47-57 of SEQ ID NO: 14), QXXXXX (aa 47-52 of SEQ ID NO: 14), QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7 or 8 or 9 or 25), or QDISNY (aa 47-52 of SEQ ID NO: 10 or 11 or 12 or 27), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 7-12;

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or further consisting of a sequence selected from the group: XXS (aa 75-77 of SEQ ID NO: 14), LVS (aa 75-77 of SEQ ID NO: 7 or 8 or 9 or 25), or YTS (aa 70-72 of SEQ ID NO: 10 or 11 or 12 or 27), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 7-12; and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising, or alternatively consisting essentially of, or further consisting of, a sequence selected from the group: XQGXXXXXT (aa 114-122 of SEQ ID NO: 14), WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25), or QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 7-12,

optionally, wherein the antibody or fragment thereof comprises or alternatively consists essentially of or additionally consists of a light chain and a heavy chain, further optionally, wherein the light chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical, to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one, two, or three CDRs of the antibody or fragment thereof, and further optionally, wherein the heavy chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one or two or three CDRs of the antibody or fragment thereof.

Embodiment 134. An antibody or fragment thereof, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7 or 8 or 9 or 25);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVS (aa 75-77 of SEQ ID NO: 7 or 8 or 9 or 25); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFP (aa 114-120 of SEQ ID NO: 7 or 8 or 9 or 25),

optionally, wherein the antibody or fragment thereof comprises or alternatively consists essentially of or additionally consists of a light chain and a heavy chain, further optionally, wherein the light chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical, to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one, two, or three CDRs of the antibody or fragment thereof, and further optionally, wherein the heavy chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one or two or three CDRs of the antibody or fragment thereof.

Embodiment 135. The antibody or fragment thereof according to any of embodiments 133-134, characterized in that CDRH1 comprises, consists essentially of, or additionally consists of the sequence GFTFRTYA (aa 50-57 of SEQ ID NO: 1 or 2 or 3 or 24).

Embodiment 136. The antibody or fragment thereof according to any of embodiments 133-135, characterized in that CDRH1 comprises, consists essentially of, or additionally consists of the sequence aASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 24), wherein the lowercase letter a is A (aa 47-59 of SEQ ID NO: 1 or 2) or wherein the lowercase letter a is K (aa 47-59 of SEQ ID NO: 3).

Embodiment 137. The antibody or fragment thereof according to any of embodiments 133-136, characterized in that CDRH2 comprises, consists essentially of, the sequence IGSDRRHT (aa 75-81 of SEQ ID NO: 1 or 2 or 3 or 24).

Embodiment 138. The antibody or fragment thereof according to any of embodiments 133-137, wherein CDRH2 comprises, consists essentially of, or additionally consists of the sequence IGSDRRHTY (aa 75-83 of SEQ ID NO: 1 or 2 or 3 or 24).

Embodiment 139. The antibody or fragment thereof according to any of embodiments 133-138, wherein CDRH2 comprises, consists essentially of, or additionally consists of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24).

Embodiment 140. The antibody or fragment thereof according to any of embodiments 133-139, characterized in that CDRH2 comprises or consists essentially of or additionally consists of the sequence WVATIGSDRRHTYYP (aa 71-85 of SEQ ID NO: 1 or 2 or 3 or 24).

Embodiment 141. The antibody or fragment thereof according to any of embodiments 133-140, characterized in that CDRL1 comprises, consists essentially of, or additionally consists of the sequence rSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 25), wherein the lowercase letter r is R (aa 44-59 of SEQ ID NO: 7 or 8) or wherein the lowercase letter r is K (aa 44-59 of SEQ ID NO: 9).

Embodiment 142. The antibody or fragment thereof according to any of embodiments 133-141, characterized in that CDRL2 comprises, consists essentially of, or additionally consists of the sequence LVSKID (aa 75-81 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 75-81 of SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aa 75-81 of SEQ ID NO: 8).

Embodiment 143. The antibody or fragment thereof according to any of embodiments 133-142, characterized in that CDRL2 comprises, consists essentially of, the sequence YLVSK1DS (aa 74-81 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 74-81 of SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aa 74-81 of SEQ ID NO: 8).

Embodiment 144. The antibody or fragment thereof according to any of embodiments 133-142, characterized in that CDRL2 comprises, consists essentially of, the sequence LVSK1DSG (aa 75-81 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 75-82 of SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aa 75-82 of SEQ ID NO: 8).

Embodiment 145. The antibody or fragment thereof according to any of embodiments 133-144, characterized in that CDRL2 comprises, consists essentially of, the sequence YLVSK1DSGV (aa 74-83 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 74-83 of SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aa 74-83 of SEQ ID NO: 8).

Embodiment 146. The antibody or fragment thereof according to any of embodiments 133-145, characterized in that CDRL2 comprises, consists essentially of, the sequence RLIYLVSK1DSGVPD (aa 71-85 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 71-85 of SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aa 71-85 of SEQ ID NO: 8).

Embodiment 147. The antibody or fragment thereof according to any one of embodiments 134-146, characterized in that CDRL3 comprises or essentially consists of or additionally consists of the sequence WQGTHFPY (aa 114-121 SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 148. The antibody or fragment thereof according to any one of embodiments 133-147, characterized in that CDRL3 comprises, consists essentially of, or additionally consists of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 149. An antibody or fragment thereof, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7 or 8 or 9 or 25);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVS (aa 75-77 of SEQ ID NO: 7 or 8 or 9 or 25); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 150. An antibody or fragment thereof according to any of embodiments 133, 134 and 149, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTYA (aa 50-57 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence IGSDRRHT (aa 75-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7 or 8 or 9 or 25);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVS (aa 75-77 of SEQ ID NO: 7 or 8 or 9 or 25); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 151. An antibody or fragment thereof according to any of embodiments 133, 134 and 149, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence aASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 24), wherein the lowercase letter a is A (aa 47-59 of SEQ ID NO: 1 or 2) or wherein the lowercase letter a is K (aa 47-59 of SEQ ID NO: 3);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence rSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 25), wherein the lowercase r is R (aa 44-59 of SEQ ID NO: 7 or 8), or wherein the lowercase r is K (aa 44-59 of SEQ ID NO: 9);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting primarily of or further consisting of the sequence YLVSKID (aa 74-81 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 74-81 of SEQ ID NO: 7 or 9), or wherein the lowercase letter l is R (aa 74-8 of SEQ ID NO: 8); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 152. An antibody or fragment thereof according to any of embodiments 133, 134, 149 and 151, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence AASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 1 or 2);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively essentially consisting of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RSSQSLLDSDGKTFLN aa (44-59 SEQ ID NO: 7);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YLVSKLDS (aa 74-81 of SEQ ID NO: 7); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 153. An antibody or fragment thereof according to any of embodiments 133, 134, 149 and 151, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence AASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 1 or 2);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 8);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YLVSKRDS (aa 74-81 of SEQ ID NO: 8); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 154. An antibody or fragment thereof according to any of embodiments 133, 134, 149 and 151, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence AASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 1 or 2);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence KSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 9);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YLVSKLDS (aa 74-81 of SEQ ID NO: 9); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 155. The antibody or fragment thereof according to any of embodiments 133, 134, 149 and 151, comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence KASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 3);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 7);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YLVSKLDS (aa 74-8 of SEQ ID NO: 7); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 156. An antibody or fragment thereof according to any of embodiments 133, 134, 149 and 151, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence KASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 3);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 8);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YLVSKRDS (aa 74-81 of SEQ ID NO: 8); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 157. An antibody or fragment thereof according to any of embodiments 133, 134, 149, 151, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence KASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 3);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TIGSDRRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence KSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 9);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YLVSKLDS (aa 74-81 of SEQ ID NO: 9); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 158. An antibody or fragment thereof according to any of embodiments 133, 134 or 149, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence rSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 25), wherein the lowercase r is R (aa 44-59 of SEQ ID NO: 7 or 8), or wherein the lowercase r is K (aa 44-59 of SEQ ID NO: 9);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVSKID (aa 75-81 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 75-81 of SEQ ID NO: 7 or 9), or wherein the lowercase letter l is R (aa 7-81 of SEQ ID NO: 8); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 159. An antibody or fragment thereof according to any of embodiments 133, 134, 149 or 158, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 7);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVSKLDS (aa 75-81 of SEQ ID NO: 7); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 160. An antibody or fragment thereof according to any of embodiments 133, 134, 149 or 158, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 8);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVSKRDS (aa 75-81 of SEQ ID NO: 8); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 161. An antibody or fragment thereof according to any of embodiments 133, 134, 149 or 158, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence KSSQSLLDSDGKTFLN (aa 44-59 of SEQ ID NO: 9);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVSKLDS (aa 75-81 of SEQ ID NO: 9); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 162. An antibody or fragment thereof according to embodiment 133 or 134, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFRTYA (aa 50-57 of SEQ ID NO: 1 or 2 or 3 or 24);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence IGSDRRHT (aa 75-81 of SEQ ID NO: 1 or 2 or 3 or 24);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7 or 8 or 9 or 25);

(v) a light chain complementarity determining region 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence LVS (aa 75-77 of SEQ ID NO: 7 or 8 or 9 or 25); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence WQGTHFP (aa 114-120 of SEQ ID NO: 7 or 8 or 9 or 25).

Embodiment 163. An antibody or fragment thereof, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively primarily consisting of or additionally consisting of the sequence GFTFSRY (aa 50-56 of SEQ ID NO: 4 or 5 or 6 or 26);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence SSGGSY (aa 76-81 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of an ER sequence (aa 121-122 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QDISNY (aa 47-52 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of a YTS sequence (aa 70-72 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQ (aa 109-110 of SEQ ID NO: 10 or 11 or 12 or 27),

optionally, wherein the antibody or fragment thereof comprises or alternatively consists essentially of or additionally consists of a light chain and a heavy chain, further optionally, wherein the light chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical, to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one, two, or three CDRs of the antibody or fragment thereof, and further optionally, wherein the heavy chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one or two or three CDRs of the antibody or fragment thereof.

Embodiment 164. The antibody or fragment thereof according to embodiment 133 or 163, characterized in that CDRH1 comprises, consists essentially of, or additionally consists of the sequence GFTFSRYG (aa 50-57 of SEQ ID NO: 4 or 5 or 6 or 26).

Embodiment 165. The antibody or fragment thereof according to any of embodiments 133 or 163-164, characterized in that CDRH1 comprises, consists essentially of, or additionally consists of the sequence aASGFTFSRYGMS (aa 47-59 of SEQ ID NO: 26), wherein the lowercase letter a is A (aa 47-57 of SEQ ID NO: 4 or 5) or wherein the lowercase letter a is T (aa 47-57 of SEQ ID NO: 6).

Embodiment 166. The antibody or fragment thereof according to any of embodiments 133 or 163-165, characterized in that CDRH2 comprises, consists essentially of, or additionally consists of the sequence ISSGGSYT (aa 75-81 of SEQ ID NO: 4 or 5 or 6 or 26).

Embodiment 167. The antibody or fragment thereof according to any of embodiments 133 or 163-166, characterized in that CDRH2 comprises, consists essentially of, or additionally consists of the sequence TISSGGSYTY (aa 74-83 of SEQ ID NO: 4 or 5 or 6 or 26).

Embodiment 168. The antibody or fragment thereof according to any of embodiments 133 or 163-167, characterized in that CDRH3 comprises, consists essentially of, or additionally consists of the sequence ERHGGDGYWYFDV (aa 121-133 SEQ ID NO: 4 or 5 or 6 or 26).

Embodiment 169. The antibody or fragment thereof according to any of embodiments 133 or 163-168, characterized in that CDRL1 comprises, consists essentially of, or additionally consists of the sequence RASQDISNYLN (aa 44-54 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 170. The antibody or fragment thereof according to any of embodiments 133 or 163-169, characterized in that CDRL2 comprises, consists essentially of, or additionally consists of the sequence YTSRLHS (aa 70-76 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 171. The antibody or fragment thereof according to any of embodiments 133 or 163-170, wherein CDRL2 comprises, consists essentially of, the sequence YYTSRLHS (aa 69-76 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 172. The antibody or fragment thereof according to any of embodiments 133 or 163-171, characterized in that CDRL3 comprises, consists essentially of, or additionally consists of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 173. An antibody or fragment thereof according to embodiment 133 or 163, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFSRY (aa 50-56 of SEQ ID NO: 4 or 5 or 6 or 26);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence SSGGSY (aa 76-81 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QDISNY (aa 47-52 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of a YTS sequence (aa 70-72 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 174. An antibody or fragment thereof according to any of embodiments 133, 163 or 173, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFSRYG (aa 50-57 of SEQ ID NO: 4 or 5 or 6 or 26);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence ISSGGSYT (aa 75-81 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QDISNY (aa 47-52 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of a YTS sequence (aa 70-72 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 175. An antibody or fragment thereof according to any of embodiments 133, 163 or 173, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence aASGFTFSRYGMS (aa 47-59 of SEQ ID NO: 26), wherein the lowercase letter a is A (aa 47-57 of SEQ ID NO: 4 or 5) or wherein the lowercase letter a is T (aa 47-57 of SEQ ID NO: 6);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TISSGGSYTY (aa 74-83 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RASQDISNYLN (aa 44-54 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YYTSRLHS (aa 69-76 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 176. An antibody or fragment thereof according to any of embodiments 133, 163, 173 or 175, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence AASGFTFSRYGMS (aa 47-57 of SEQ ID NO: 4 or 5);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TISSGGSYTY (aa 74-83 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RASQDISNYLN (aa 44-54 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YYTSRLHS (aa 69-76 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 177. An antibody or fragment thereof according to any of embodiments 133, 163, 173 or 175, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence TASGFTFSRYGMS (aa 47-57 of SEQ ID NO: 6);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence TISSGGSYTY (aa 74-83 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RASQDISNYLN (aa 44-54 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YYTSRLHS (aa 69-76 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 178. An antibody or fragment thereof according to any of embodiments 133, 163 or 173, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFSRY (aa 50-56 of SEQ ID NO: 4 or 5 or 6 or 26);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence SSGGSY (aa 76-81 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of the sequence ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence RASQDISNYLN (aa 44-54 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of the sequence YTSRLHS (aa 70-76 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 179. An antibody or fragment thereof according to any of embodiments 133 or 163, which comprises or alternatively consists essentially of or additionally consists of one, two, three, four, five or all six of the following regions (i) to (vi):

(i) a heavy chain complementarity determining region 1 (CDRH1) comprising or alternatively consisting essentially of or additionally consisting of the sequence GFTFSRYG (aa 50-57 of SEQ ID NO: 4 or 5 or 6 or 26);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of the sequence ISSGGSYT (aa 75-81 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of an ER sequence (aa 121-122 of SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a complementarity determining region of the light chain 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of the sequence QDISNY (aa 47-52 of SEQ ID NO: 10 or 11 or 12 or 27);

(v) a complementarity determining region of the light chain 2 (CDRL2) comprising or alternatively consisting essentially of or additionally consisting of a YTS sequence (aa 70-72 of SEQ ID NO: 10 or 11 or 12 or 27); and

(vi) a complementarity determining region of the light chain 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of the sequence QQ (aa 109-110 of SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 180. An antibody or fragment thereof according to any one of embodiments 1-179, characterized in that said antibody comprises or alternatively consists essentially of or additionally consists of:

(i) CDRs 1-3 of a sequence selected from SEQ ID NO: 1-6, 13, 24 or 26, or an equivalent of each; and/or

(ii) CDRs 1-3 of a sequence selected from SEQ ID NO: 7-12, 14, 25 or 27, or an equivalent of each.

Embodiment 181. An antibody or fragment thereof according to any one of embodiments 1-180, characterized in that said antibody comprises, or alternatively consists essentially of, or additionally consists of:

(i) CDRs 1-3 of a sequence selected from SEQ ID NO: 1-6, 13, 24 or 26; and/or

(ii) CDRs 1-3 of a sequence selected from SEQ ID NO: 7-12, 14, 25 or 27.

Embodiment 182. An antibody or fragment thereof, comprising or alternatively consisting essentially of or additionally consisting of:

(i) CDRs 1-3 of a sequence selected from the group of SEQ ID NOs: 1-6, 13, 24 or 26, or the equivalent of each of them; and/or

(ii) CDRs 1-3 of a sequence selected from the group of SEQ ID NOs: 7-12, 14, 25 or 27, or an equivalent of each of them,

optionally, wherein the antibody or fragment thereof comprises or alternatively consists essentially of or additionally consists of a light chain and a heavy chain, further optionally, wherein the light chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical, to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one, two, or three CDRs of the antibody or fragment thereof, and further optionally, wherein the heavy chain is at least about 80% identical, or at least about 85% identical, or at least about 90% identical, or at least about 91% identical, or at least about 92% identical, or at least about 93% identical, or at least about 94% identical, or at least about 95% identical, or at least about 96% identical, or at least about 97% identical, or at least about 98% identical, or at least about 99% identical to the amino acid sequence of any one or more of SEQ ID NOs: 7-12, 14, 25, or 27, or optionally selected from SEQ ID NOs, the sequence of which comprises one or two or three CDRs of the antibody or fragment thereof.

Embodiment 183. An antibody or fragment thereof according to any one of embodiments 1-182, characterized in that the antibody is selected from the group consisting of a bispecific antibody, a trispecific antibody, a tetraspecific antibody or a pentaspecific antibody.

Embodiment 184. An antibody or fragment thereof according to any one of embodiments 1-183, characterized in that the antibody is selected from the group of IgA, IgD, IgE, IgG or IgM antibodies.

Embodiment 185. An antibody or fragment thereof according to any one of embodiments 1-184, characterized in that the antibody further comprises a constant region selected from the group: an IgA constant region, an IgD constant region, an IgE constant region, an IgG constant region, or an IgM constant region.

Embodiment 186. An antibody or fragment thereof according to embodiment 185, wherein the constant region is an IgG1 constant region.

Embodiment 187. An antibody or fragment thereof according to any one of embodiments 1-185, characterized in that said antibody further comprises a heavy chain constant region (HC) of SEQ ID NO: 1-6, 13, 24 or 26 and/or a light chain constant region (LC) of SEQ ID NO: 7-12, 14, 25 or 27.

Embodiment 188. The antibody or fragment thereof according to embodiment 187, characterized in that the HC constant region comprises or alternatively essentially consists of or additionally consists of a constant region of SEQ ID NO: 1-6, 13, 24 or 26 (optionally a sequence selected from aa 145-473 of SEQ ID NO: 1-6, 13, 24 or 26), or characterized in that the HC constant regions comprise or alternatively essentially consist of or additionally consist of a constant region of any of SEQ ID NO: 15-22.

Embodiment 189. The antibody or fragment thereof according to embodiment 187 or 188, characterized in that the LC constant region comprises the constant region of SEQ ID NO: 7-12 or 27 (optionally the sequence is selected from aa 133-239 of SEQ ID NO: 7-9, 14 or 25, and/or aa 127-233 of SEQ ID NO: 10-12 or 27), or characterized in that the LC constant regions comprise or alternatively essentially consist of or additionally consist of the constant region of SEQ ID NO: 23.

Embodiment 190. An antibody or fragment thereof that competes for epitope binding with an antibody or fragment thereof according to any one of embodiments 1-189.

Embodiment 191. An antibody or fragment thereof that competes for binding of the head chimeric peptide IhfA5-mIhfB4NTHI with an antibody or fragment thereof according to any one of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-189.

Embodiment 192. The antibody or fragment thereof according to embodiment 191, wherein the IhfA5-mIhfB4NTHI head chimeric peptide comprises or alternatively consists essentially of or additionally consists of RPGRNPX1TGDVVPVSARRVV-X-FSLHHRQPRLGRNPX1TGDSV (SEQ ID NO: 38), wherein "X" is an optional amino acid linker sequence, optionally comprising or consisting essentially of or additionally consisting of from 1 to 20 amino acids, and wherein "X1" is any amino acid or alternatively "X1" is selected from the amino acids Q, R, K, C or T.

Embodiment 193. The antibody or fragment thereof according to embodiment 191 or 192, characterized in that the head chimeric peptide IhfA5-mIhfB4NTHI comprises or alternatively consists essentially of or additionally consists of RPGRNPKTGDVVPVSARRVV-X-FSLHHRQPRLGRNPKTGDSV (SEQ ID NO: 39), wherein "X" is an optional amino acid linker sequence, optionally comprising or consisting essentially of or additionally consisting of from 1 to 20 amino acids.

Embodiment 194. The antibody or fragment thereof according to any of embodiments 191-193, characterized in that the head chimeric peptide IhfA5-mIhfB4NTHI comprises or alternatively essentially consists of or additionally consists of RPGRNPKTGDVVPVSARRVVGPSLFSLHHRQPRLGRNPKTGDSV (SEQ ID NO: 40).

Embodiment 195. An antibody or fragment thereof that competes for binding of the IhfA3-IhfB2NTHI tail chimeric peptide with an antibody or fragment thereof according to any one of embodiments 1, 3-15, 25-34, 36-47, 58-67, 69-81, 91-100, 102-114, 124-133 and 163-189.

Embodiment 196. The antibody or fragment thereof according to embodiment 195, characterized in that the IhfA3-IhfB2NTHI tail chimeric peptide comprises or alternatively consists essentially of or additionally consists of FLEEIRLSLESGQDVKLSGF-X-TLSAKEEIENMVKDILEFISQ (SEQ ID NO: 41), wherein "X" is an optional amino acid linker sequence, optionally comprising or consisting essentially of or additionally consisting of from 1 to 20 amino acids, and/or wherein the IhfA3-IhfB2NTHI tail chimeric peptide comprises or consists essentially of or additionally consists of FLEEIRLSLESGQDVKLSGFGPSLTLSAKEIENMVKDILEFISQ (SEQ ID NO: 50).

Embodiment 197. The antibody or fragment thereof according to any of embodiments 192, 193 or 196, characterized in that the amino acid linker is selected from the group consisting of: GGSGGS (SEQ ID NO: 42), GPSLKL (SEQ ID NO: 43), GGG (SEQ ID NO: 44), GPSL (SEQ ID NO: 45), GPS (SEQ ID NO: 46), PSLK (SEQ ID NO: 47), GPSLK (SEQ ID NO: 48) or SLKL (SEQ ID NO: 49).

Embodiment 198. An antibody or fragment thereof according to any one of embodiments 190-197, characterized in that the antibody is a polyclonal, monoclonal or humanized antibody.

Embodiment 199. An antigen-binding fragment of an antibody according to any one of embodiments 1-198.

Embodiment 200. An antigen-binding fragment according to embodiment 199, characterized in that the antigen-binding fragment is selected from the group of Fab, F(ab’)2, Fab’, scFv or Fv.

Embodiment 201. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 80% amino acid identity with the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 202. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 80% amino acid identity with the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 203. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 95% amino acid identity with the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 204. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 96% amino acid identity with the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 205. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 97% amino acid identity with the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 206. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 98% amino acid identity with the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 207. An antibody or fragment thereof according to any of embodiments 1-33, 67-132 or 180-198, or an antigen-binding fragment according to embodiment 199 or 200, characterized in that the amino acid sequence equivalent comprises a polypeptide having at least 99% amino acid identity to the amino acid sequence, and/or characterized in that the amino acid sequence equivalent comprises a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complementary sequence of a polynucleotide encoding the amino acid sequence.

Embodiment 208. An antibody or fragment thereof according to any of embodiments 1-198 and 201-207, or an antigen-binding fragment according to any of embodiments 199-207, characterized in that the antibody or fragment thereof is modified and, optionally, characterized in that the modification is selected from the group of PEGylation, PEG mimetic, polysialylation, HESylation or glycosylation.

Embodiment 209. An antibody or fragment thereof according to any of embodiments 1-198 and 201-208, or an antigen-binding fragment according to any of embodiments 199-208, further comprising a detectable marker or a purification marker.

Embodiment 210. The complementarity determining region (CDR) of an antibody or fragment thereof according to any of embodiments 1-198 and 201-209, or the antigen-binding fragment according to any of embodiments 199-209.

Embodiment 211. A CDR according to embodiment 210, wherein the CDR is selected from the group consisting of CDR1, CDR2 or CDR3.

Embodiment 212. The complementarity determining region (CDR) comprises or alternatively consists essentially of or additionally consists of any one or more of the following regions:

(i) a complementarity determining region (CDR), namely a heavy chain complementarity determining region 1 (CDRH1), comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: GFTFXXY (amino acids (aa) 50-56 of SEQ ID NO: 13), GFTFRTY (aa 50-56 of SEQ ID NO: 1 or 2 or 3 or 24), or GFTFSRY (aa 50-56 of SEQ ID NO: 4 or 5 or 6 or 26), characterized in that X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 1-6; GFTFRTYA (aa 50-57 of SEQ ID NO: 1 or 2 or 3 or 24); aASGFTFRTYAMS (aa 47-59 of SEQ ID NO: 24), wherein the lowercase letter a is A (aa 47-59 of SEQ ID NO: 1 or 2) or wherein the lowercase letter a is K (aa 47-59 of SEQ ID NO: 3); GFTFSRYG (aa 50-57 of SEQ ID NO: 4 or 5 or 6 or 26); or aASGFTFSRYGMS (aa 47-59 of SEQ ID NO: 26), wherein the lowercase letter a is A (aa 47-57 of SEQ ID NO: 4 or 5) or wherein the lowercase letter a is T (aa 47-57 of SEQ ID NO: 6);

(ii) a heavy chain complementarity determining region 2 (CDRH2) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: XSXXXX (amino acids (aa) 76-81 of SEQ ID NO: 13), GSDRRH (aa 76-81 of SEQ ID NO: 1 or 2 or 3 or 24), or SSGGSY (aa 76-81 of SEQ ID NO: 4 or 5 or 6 or 26), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 1-6; IGSDRRHT (aa 75-81 of SEQ ID NO: 1 or 2 or 3 or 24); IGSDDRHTY (aa 75-83 of SEQ ID NO: 1 or 2 or 3 or 24); TIGSDDRHTY (aa 74-83 of SEQ ID NO: 1 or 2 or 3 or 24); WVATIGSDRRHTYYP (aa 71-85 of SEQ ID NO: 1 or 2 or 3 or 24); ISSGGSYT (aa 75-81 of SEQ ID NO: 4 or 5 or 6 or 26); or TISSGGSYTY (aa 74-83 of SEQ ID NO: 4 or 5 or 6 or 26);

(iii) a heavy chain complementarity determining region 3 (CDRH3) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: XXXXXXXYXXFDX (amino acids (aa) 121-133 of SEQ ID NO: 13), VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24), or ERHGGDGYWYFDV (aa 121-133 of SEQ ID NO: 4 or 5 or 6 or 26), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 1-6; VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1 or 2 or 3 or 24); or ER (ak 121-122 SEQ ID NO: 4 or 5 or 6 or 26);

(iv) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: QXXXXXXXXXX (aa 47-52 of SEQ ID NO: 14), QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7 or 8 or 9 or 25), or QDISNY (aa 47-52 of SEQ ID NO: 10 or 11 or 12 or 27), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 7-12; rSSQSLLDSDGKTFLN (ac 44-59 SEQ ID NO: 25), where the lowercase r is R (ac 44-59 SEQ ID NO: 7 or 8), or where the lowercase r is K (ac 44-59 SEQ ID NO: 9); or RASQDISNYLN (ac 44-54 SEQ ID NO: 10 or 11 or 12 or 27);

(v) a light chain complementarity determining region 1 (CDRL1) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: XXS (aa 75-77 of SEQ ID NO: 14), LVS (aa 75-77 of SEQ ID NO: 7 or 8 or 9 or 25), or YTS (aa 70-72 of SEQ ID NO: 10) or 11 or 12 or 27), wherein X is any amino acid or an amino acid in an aligned position of a sequence selected from SEQ ID NO: 7-12; LVSKlDS (aa 75-81 of SEQ ID NO: 25), wherein the lowercase letter l is L (aa 75-81 of SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aa 75-81 of SEQ ID NO: 8); YLVSKlDS (ac 74-81 SEQ ID NO: 25), where the lowercase letter l is L (ac 74-81 SEQ ID NO: 7 or 9) or where the lowercase letter l is R (ac 74-81 SEQ ID NO: 8); LVSKIDSG (ac 75-82 SEQ ID NO: 25), where the lowercase letter l is L (ac 75-82 SEQ ID NO: 7 or 9) or where the lowercase letter l is R (ac 75-82 SEQ ID NO: 8); YLVSKIDSGV (ac 74-83 SEQ ID NO: 25), where the lowercase letter l is L (ac 74-83 SEQ ID NO: 7 or 9) or where the lowercase letter l is R (ac 74-83 SEQ ID NO: 8); RLIYLVSKIDSGVPD (aca 71-85 SEQ ID NO: 25), wherein the lowercase letter l is L (aca 71-85 SEQ ID NO: 7 or 9) or wherein the lowercase letter l is R (aca 71-85 SEQ ID NO: 8); YTSRLHS (aca 70-76 SEQ ID NO: 10, or 11, or 12, or 27); or YYTSRLHS (aca 69-76 SEQ ID NO: 10, or 11, or 12, or 27); and

(vi) a light chain complementarity determining region 3 (CDRL3) comprising or alternatively consisting essentially of or additionally consisting of a sequence selected from the group: XQGXXXXXT (aa 114-122 of SEQ ID NO: 14), WQGTHFPYT (aa 114-122 of SEQ ID NO: 7 or 8 or 9 or 25), or QQGNPLRT (aa 109-116 of SEQ ID NO: 10 or 11 or 12 or 27), wherein X is any amino acid or an amino acid at an aligned amino acid position of a sequence selected from SEQ ID NO: 7-12; WQGTHFP (aa 114-120 of SEQ ID NO: 7 or 8 or 9 or 25); WQGTHFPY (ac 114-121 SEQ ID NO: 7 or 8 or 9 or 25); WQGTHFPYT (ac 114-122 SEQ ID NO: 7 or 8 or 9 or 25); QQ (ac 109-110 SEQ ID NO: 10 or 11 or 12 or 27).

Embodiment 213. An isolated polypeptide comprising an amino acid sequence selected from the group of SEQ ID NOs: 1-14 or 24-27, or an equivalent of each.

Embodiment 214. An isolated polypeptide according to embodiment 213, wherein the polypeptide further comprises a detectable marker or a purification marker.

Embodiment 215. An isolated polynucleotide encoding an antibody or fragment thereof according to any of embodiments 1-198 and 201-209, an antigen-binding fragment according to any of embodiments 199-209, a CDR according to any of embodiments 210-212, and an isolated polypeptide according to embodiment 213 or 214, or an equivalent of each, and optionally operably linked to a promoter and an enhancer element.

Embodiment 216. An isolated polynucleotide sequence according to embodiment 215, further comprising a polynucleotide sequence encoding a signal peptide located upstream of the variable domain, chain or CDR.

Embodiment 217. An antibody or fragment thereof according to any of embodiments 1-198 and 201-209, an antigen-binding fragment according to any of embodiments 199-209, a CDR according to any of embodiments 210-212 and an isolated polypeptide according to embodiment 213 or 214, further comprising a signal peptide located upstream of the variable domain, chains or CDRs.

Embodiment 218. A vector comprising an isolated polynucleotide according to embodiment 215 or 216.

Embodiment 219. The vector of embodiment 218, wherein the vector is a plasmid or a viral vector.

Embodiment 220. The vector of embodiment 219, wherein the viral vector is selected from the group consisting of a retroviral vector, a lentiviral vector, an adenoviral vector, and an adeno-associated virus vector.

Embodiment 221. A host cell comprising a polynucleotide according to embodiment 215 or 216 or a vector according to any of embodiments 218-220.

Embodiment 222. A composition comprising a carrier and one or more compounds selected from the group consisting of an antibody or fragment thereof according to any of embodiments 1-198, 201-209 and 217, an antigen-binding fragment according to any of embodiments 199-209 and 217, a CDR according to any of embodiments 210-212 and 217, an isolated polypeptide according to embodiment 213 or 214, an isolated polynucleotide according to embodiment 215 or 216, a vector according to any of embodiments 218-220 or a host cell according to embodiment 221.

Embodiment 223. A method for producing an antibody or fragment thereof according to any one of embodiments 1-198, 201-209 and 217, comprising culturing a host cell containing a polynucleotide encoding the antibody or fragment thereof under conditions allowing expression of the antibody or fragment thereof, and optionally isolating the antibody or fragment thereof.

Embodiment 224. The method of embodiment 223, wherein the host cell is a mammalian cell.

Embodiment 225. A method for inhibiting or competing for the binding of a DNABII polypeptide or protein to microbial DNA, comprising contacting the DNABII polypeptide or protein with an antibody or fragment thereof according to any of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, characterized in that the antibody or fragment thereof binds to the head region of the DNABII peptide.

Embodiment 226. A method for disrupting a biofilm, comprising contacting the biofilm with an antibody or fragment thereof according to any one of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, characterized in that the antibody or fragment thereof binds to the head region of the DNABII peptide.

Embodiment 227. A method for preventing the formation or destruction of a biofilm on a surface, comprising treating a surface susceptible to or containing a biofilm with an antibody or fragment thereof according to any one of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, characterized in that the antibody or fragment thereof binds to the head region of the DNABII peptide.

Embodiment 228. A method for preventing or destroying a biofilm in a subject, comprising administering to the subject an antibody or fragment thereof according to any one of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, characterized in that the antibody or fragment thereof binds to the head region of the DNABII peptide.

Embodiment 229. A method for inhibiting, preventing or treating a microbial infection that causes biofilm formation in a subject, comprising administering to the subject an antibody or fragment thereof according to any one of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, characterized in that the antibody or fragment thereof binds to the head region of the DNABII peptide.

Embodiment 230. A method for treating a condition characterized by biofilm formation in a subject, comprising administering to the subject an antibody or fragment thereof according to any one of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, characterized in that the antibody or fragment thereof binds to the head region of the DNABII peptide.

Embodiment 231. The method according to any one of embodiments 228-230, characterized in that the antibody or fragment thereof reduces the level of one or more pro-inflammatory cytokines and increases the level of one or more anti-inflammatory cytokines in the subject.

Embodiment 232. The method of embodiment 230 or 231, wherein the condition is selected from the group consisting of chronic non-healing wounds, infections caused by Burkholderia sp., venous ulcers, diabetic foot ulcers, ear infections, sinus infections, urinary tract infections, gastrointestinal diseases, pulmonary infections, respiratory tract infections, cystic fibrosis, chronic obstructive pulmonary disease, catheter-related infections, indwelling device-related infections, implanted prosthetic infections, osteomyelitis, cellulitis, abscesses, and periodontal disease.

Embodiment 233. The method according to any one of embodiments 225-232, further comprising detecting a biofilm by contacting an antibody that binds a DNABII polypeptide or an antigen-binding fragment of an antibody with a sample suspected of containing a biofilm, and detecting binding of the biofilm and the antibody or fragment thereof.

Embodiment 234. A method for detecting a biofilm in a subject, comprising administering to the subject an antibody or fragment thereof according to any one of embodiments 1-198, 201-209 and 217 and detecting binding of the antibody or fragment thereof to the biofilm.

Embodiment 235. A method for detecting a microbial infection causing biofilm formation in a subject, comprising contacting an antibody or fragment thereof according to any of embodiments 1-198, 201-209 and 217 with a biological sample suspected of containing a biofilm and isolated from the subject's body and detecting binding of the antibody or fragment thereof to any biofilm in the sample.

Embodiment 236. A method for screening subjects having a biofilm, comprising contacting an antibody or fragment thereof according to any one of embodiments 1-198, 201-209 and 217 with a biological sample containing a biofilm and isolated from the subject's body, and detecting binding of the antibody or fragment thereof to any biofilm in the sample.

Embodiment 237. A method for generating passive immunity in a subject, comprising administering to the subject an antibody or fragment thereof according to any of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, which binds to the head region of the DNABII peptide.

Embodiment 238. The method according to any one of embodiments 225-237, characterized in that the biofilm is obtained from Gram-negative or Gram-positive biofilm-producing bacteria.

Embodiment 239. The method according to any one of embodiments 225-238, characterized in that the biofilm comprises the DNABII protein.

Embodiment 240. The method according to embodiment 239, characterized in that the biofilm comprises a histone-like protein from the E. coli strain U93 (HU) or a host integration factor (IHF) binding protein.

Embodiment 241. A method for producing an interfering nucleic acid, comprising obtaining a nucleic acid consisting of about 10-20 nucleotides that specifically binds a binding partner of an antibody or fragment thereof according to any of embodiments 1-198, 201-209 and 217, and optionally isolating the obtained interfering nucleic acid.

Embodiment 242. A non-physiological surface coated with an antibody or fragment thereof according to any of embodiments 1-198, 201-209 and 217, wherein the surface is optionally under industrial conditions.

Embodiment 243. A method for producing an antiserum effective for destroying a biofilm, comprising immunizing a subject with a small molecule and obtaining the antiserum from the subject's body and, optionally, isolating a polyclonal antiserum or monoclonal antibodies from the subject's body.

Embodiment 244. An antibody fragment according to any of embodiments 1-24, 34-57, 67-90, 100-123, 133-162 and 180-198, 201-209 and 217, wherein the fragment is selected from the group of Fab, F(ab')2, Fab', scFv or Fv, and wherein the antibody fragment specifically binds to the head region of the DNABII peptide.

Embodiment 245. The antibody or fragment thereof according to embodiment 244 or the method according to any of embodiments 225-230 and 70-74, characterized in that the DNABII peptide is the IHF peptide.

Embodiment 246. A composition comprising an antibody fragment according to embodiment 244 or 245 and a pharmaceutically acceptable carrier.

Embodiment 247. A non-physiological surface coated with an antibody fragment according to embodiment 244 or 245, and optionally wherein the surface is under industrial conditions.

Embodiment 248. A kit comprising one or more of the following: an antibody or fragment thereof according to any of embodiments 1-198, 201-209, 217, 244 and 245, an antigen-binding fragment according to any of embodiments 199-209, a CDR of any of embodiments 210-212 and 217, an isolated polypeptide according to embodiment 213 or 214, a polynucleotide according to embodiment 215 or 216, a vector according to any of embodiments 218-220, a host cell according to embodiment 221 or a composition according to embodiment 222 or 246; and optionally instructions for use.

SEQUENCE LIST

SEQ ID NO: (H10210 (1F8.F1 Humanized HC1))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

NHYTQKSLSLSPG**

SEQ ID NO: (H10211 (1F8.F1 Humanized HC2))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (H10212 (1F8.F1 Humanized HC3))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (H10213 (11E7.C7 Humanized HC1))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

MDPKGSLSWRILLFLSLAFELSYGEVQLVESGGGLVKPGGSLRLSCAASGTFFSRYGMSWVRQAPGKGLEWVATISSGGSYTYYTDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVY YCERHGGDGYWYFDVWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG**

SEQ ID NO: (H10214 (11E7.C7 Humanized HC2))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (H10215 (11E7.C7 Humanized HC3))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (L10210 (1F8.F1 Humanized LC1))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (L10211 (1F8.F1 Humanized LC2))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (L10212 (1F8.F1 Humanized LC3))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

SEQ ID NO: (L10213 (11E7.C7 Humanized LC1))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: (L10214 (11E7.C7 Humanized LC2))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

SEQ ID NO: (L10215 (11E7.C7 Humanized LC3))

Bold font indicates the approximate variable region, while bold, italic, and underlined font indicate the approximate CDR.

SEQ ID NO: 13 (Heavy chain consensus sequence)

MDPKGSLSWR ILFLLSLAFE LSYGEVqLVe SGgglvXPGg SlrlSCaASG 50

FTFXXYXMSW VRQAPGkgLE WVaTIXSXXX XTYYXDsvkG RfTIsRDNaK 100

NtLYlqmnSL raEDTAVYYC XXXXXXXYXX FDXWGQGTXV TVSSASTKGP 150

SVFPLAPSSK STSGGTAALG CLVKDYFPEP VTVSWNSGAL TSGVHTFPAV 200

LQSSGLYSLS SVVTVPSSSL GTQTYICNVN HKPSNTKVDK KVEPKSCDKT 250

HTCPPCPAPE LLGGPSVFLF PPKPKDTLMI SRTPEVTCVV VDVSHEDPEV 300

KFNWYVDGVE VHNAKTKPRE EQYNSTYRVV SVLTVLHQDW LNGKEYKCKV 350

SNKALPAPIE KTISKAKGQP REPQVYTLPP SREEMTKNQV SLTCLVKGFY 400

PSDIAVEWES NGQPENNYKT TPPVLDSDGS FFLYSKLTVD KSRWQQGNVF 450

SCSVMHEALH NHYTQKSLSL SPG** 475

Where X and the lowercase letter may be replaced by any amino acid or, alternatively, an amino acid selected from SEQ ID NOs: 1-6, at the corresponding position. In one embodiment, X may also indicate the absence of an amino acid residue.

SEQ ID NO: 14 (Light chain consensus sequence)

METDTLLLWV LLLWVPGSTG DXvMTQSPXs LsvslGXrat isCrXSQXXX 50

XXXXXXXLNW XQQkPGqaXX XLIYXXSXlX SGvPdRFSGS GSGTDXTLtI 100

SslXXEDXav YyCXQGXXXX XTFGXGTKXE IKRTVAAPSV FIFPPSDEQL 150

KSGTASVVCL LNNFYPREAK VQWKVDNALQ SGNSQESVTE QDSKDSTYSL 200

SSTLTLSKAD YEKHKVYACE VTHQGLSSPV TKSFNRGEC* 240

Where X and the lowercase letter may be replaced by any amino acid or, alternatively, an amino acid selected from SEQ ID NOs: 1-6, at the corresponding position. In one embodiment, X may also indicate the absence of an amino acid residue.

SEQ ID NO: 15 Human IgD constant region, Uniprot: P01880

APTKAPDVFPIISGCRHPKDNSPVVLACLITGYHPTSVTVTWYMGTQSQPQRTFPEIQRRDSYYMTSSQLSTPLQQWRQGEYKCVVQHTASKSKKE IFRWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLRDKATFTCFV VGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQHSRLTLPRSLWNAGTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAASWLLCEVSGFSPPNILLMWLEDQREVNTSGFAPARPPPQPGSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRTLLNASRSLEVSYVTDHGPMK

SEQ ID NO: 16 Human IgG1 constant region, Uniprot: P01857

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO: 17 Human IgG2 constant region, Uniprot: P01859

ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEV HNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO: 18 Human IgG3 constant region, Uniprot: P01860

ASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYTCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVQFKWYVDGVEVHNAKTKPREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKTKGQPREPQVYTLP PSREEMTKNQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGK

SEQ ID NO: 19 Human IgM constant region, Uniprot: P01871

GSASAPTLFPLVSCENSPSDTSSVAVGCLAQDFLPDSITLSWKYKNNSDISSTRGFPSVLRGGKYAATSQVLLPSKDVMQGTDEHVVCKVQHPNGNKEKNVPLPVIAELPPKV SVFVPPRDGFFGNPRKSKLICQATGFSPRQIQVSWLREGKQVGSGVTTDQVQAEAKESGPTTYKVTSTLTIKESDWLGQSMFTCRVDHRGLTFQQNASSMCVPDQDTAIRVFA IPPSFASIFLTKSTKLTCLVTDLTTYDSVTISWTRQNGEAVKTHTNISESHPNATFSAVGEASICEDDWNSGERFTCTVTHTDLPSPLKQTISRPKGVALHRPDVYLLPPARE QLNLRESATITCLVTGFSPADVFVQWMQRGQPLSPEKYVTSAPMPEPQAPGRYFAHSILTVSEEEWNTGETYTCVAHEALPNRVTERTVDKSTGKPTLYNVSLVMSDTAGTCY

SEQ ID NO: 20 Human IgG4 constant region, Uniprot: P01861

ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

SEQ ID NO: 21 Human IgA1 constant region, Uniprot: P01876

ASPTSPKVFPLSLCSTQPDGNVVIACLVQGFFPQEPLSVTWSESGQGVTARNFPPSQDASGDLYTTSSQLTLPATQCLAGKSVTCHVKHYTNPSQDVTVPCPVPSTPPTPSPSTPPTPSPSCCHPRLSLHRPALEDLLGSEANLTCTLTGLRDASGVTFTWTPSSGKSAVQGPPE RDLCGCYSVSSVLPGCAEPWNHGKTFTCTAAYPESKTPLTATLSKSGNTFRPEVHLLPPPSEELALNELVLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGDTFSCMVGHEALPLAFTQKTIDRLAGKPTHVNVSVVMAEVDGTCY

SEQ ID NO: 22 Human IgA2 constant region, Uniprot: P01877

ASPTSPKVFPLSLDSTPQDGNVVVACLVQGFFPQEPLSVTWSESGQNVTARNFPPSQDASGDLYTTSSQLTLPATQCPDGKSVTCHVKHYTNPSQDVTVPCPVPPPPPCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGATFTWTPSSGKSAVQGPPERDLCGCY SVSSVLPGCAQPWNHGETFTCTAAHPELKTPLTANITKSGNTFRPEVHLLPPPSEELALNELVLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGDTFSCMVGHEALPLAFTQKTIDRMAGKPTHVNVSVVMAEVDGTCY

SEQ ID NO: 23 Human Ig kappa constant region, Uniprot: P01834

TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO: 24 (heavy chain head consensus sequence)

MDPKGSLSWR ILFLLSLAFE LSYGEVkLVe SGgglvqPGg SlrlSCaASG 50

FTFRTYAMSW VRQAPGkgLE WVATIGSDRR HTYYPDsvkG RfTIsRDNaK 100

NTLYlqmnSL RaEDTAVYYC VGPYDGYYGE FDYWGQGTLV TVSSASTKGP 150

SVFPLAPSSK STSGGTAALG CLVKDYFPEP VTVSWNSGAL TSGVHTFPAV 200

LQSSGLYSLS SVVTVPSSSL GTQTYICNVN HKPSNTKVDK KVEPKSCDKT 250

HTCPPCPAPE LLGGPSVFLF PPKPKDTLMI SRTPEVTCVV VDVSHEDPEV 300

KFNWYVDGVE VHNAKTKPRE EQYNSTYRVV SVLTVLHQDW LNGKEYKCKV 350

SNKALPAPIE KTISKAKGQP REPQVYTLPP SREEMTKNQV SLTCLVKGFY 400

PSDIAVEWES NGQPENNYKT TPPVLDSDGS FFLYSKLTVD KSRWQQGNVF 450

SCSVMHEALH NHYTQKSLSL SPG** 475

In this case, the lowercase letter can be replaced by an amino acid selected from SEQ ID NO: 1-3 in the corresponding position.

SEQ ID NO: 25 (light chain to head consensus sequence)

METDTLLLWV LLLWVPGSTG DVVMTQSPlS LpVtLGqpAs IsCrSSQSLL 50

DSDGKTFLNW LQQrPGQsPr RLIYLVSKlD SGVPDRFSGS GSGTDFTLkI 100

SrveAEDVgV YYCWQGTHFP YTFGQGTKLE IKRTVAAPSV FIFPPSDEQL 150

KSGTASVVCL LNNFYPREAK VQWKVDNALQ SGNSQESVTE QDSKDSTYSL 200

SSTLTLSKAD YEKHKVYACE VTHQGLSSPV TKSFNRGEC** 240

In this case, the lowercase letter can be replaced by an amino acid selected from SEQ ID NO: 7-9 in the corresponding position.

SEQ ID NO: 26 (heavy chain consensus sequence to tail portion)

MDPKGSLSWR ILFLLSLAFE LSYGEVQLVE SGGGLVkPGg SLRLSCaASG 50

FTFSRYGMSW VRQAPGKGLE WVaTISSGGS YTYYTDSVKG RFTISRDNAK 100

NsLYLQMNSL raEDTAVYYC ERHGGDGYWY FDVWGQGTMV TVSSASTKGP 150

SVFPLAPSSK STSGGTAALG CLVKDYFPEP VTVSWNSGAL TSGVHTFPAV 200

LQSSGLYSLS SVVTVPSSSL GTQTYICNVN HKPSNTKVDK KVEPKSCDKT 250

HTCPPCPAPE LLGGPSVFLF PPKPKDTLMI SRTPEVTCVV VDVSHEDPEV 300

KFNWYVDGVE VHNAKTKPRE EQYNSTYRVV SVLTVLHQDW LNGKEYKCKV 350

SNKALPAPIE KTISKAKGQP REPQVYTLPP SREEMTKNQV SLTCLVKGFY 400

PSDIAVEWES NGQPENNYKT TPPVLDSDGS FFLYSKLTVD KSRWQQGNVF 450

SCSVMHEALH NHYTQKSLSL SPG** 475

In this case, the lowercase letter can be replaced by an amino acid selected from SEQ ID NO: 4-6 in the corresponding position.

SEQ ID NO: 27 (heavy chain consensus sequence to tail portion)

METDTLLLWV LLLWVPGSTG DIqMTQSPss LSaSvGdRvT itCRASQDIS 50

NYLNWYQQKP GkAVkLLIYY TSRLHSGvPs RFSGSGSGTD YTLTISSLqP 100

EDFAtYfCQQ GNPLRTFGGG TKVEIKRTVA APSVFIFPPS DEQLKSGTAS 150

VVCLLNNFYP REAKVQWKVD NALQSGNSQE SVTEQDSKDS TYSLSSTLTL 200

SKADYEKHKV YACEVTHQGL SSPVTKSFNR GEC* 234

In this case, the lowercase letter can be replaced by an amino acid selected from SEQ ID NO: 10-12 in the corresponding position.

SEQ ID NO: 28 (consensus sequence that binds to IHF)

WATCAANNNNTTR

Where W is A or T and R is purine.

SEQ ID NO. 29 (E. coli hupA, Genbank accession number: AP_003818, last accessed March 21, 2011)

MNKTQLIDVIAEKAELSKTQAKAALESTLAAITESLKEGDAVQLVGFGTFK VNHRAERTGRNPQTGKEIKIAAANVPAFVSGKALKDAVK

SEQ ID NO. 30 (E. coli hupB, Genbank accession number: AP_001090.1, last accessed March 21, 2011)

MNKSQLIDKIAAGADISKAAAGRALDAIIASVTESLKEGDDVALVGFG TFAVKERAARTGRNPQTGKEITIAAAKVPSFRAGKALKDAVN

SEQ ID NO: 31 (IhfA, A head fragment)

NFELRDKSSRPGRNPKTGDVV

SEQ ID NO: 32 (IhfB, B head fragment)

SLHHRQPRLGRNPKTGDSVNL

SEQ ID NO: 33 (peptide linker)

Gly-Pro-Ser-Leu-Lys-Leu

SEQ ID NO: 34 (peptide linker)

Gly-Pro-Ser-Leu

SEQ ID NO: 35 (peptide linker)

Pro-Ser-Leu-Lys

SEQ ID NO: 36 (peptide linker)

Gly-Pro-Ser-Leu-Lys

SEQ ID NO: 37 (peptide linker)

Ser-Leu-Lys-Leu

SEQ ID NO: 38 (IhfA5-mIhfB4NTHI head chimeric peptide)

wherein "X" is an optional amino acid linker sequence, optionally comprising or consisting essentially of or additionally consisting of from 1 to 20 amino acids

where "X1" is any amino acid or alternatively "X1" is selected from the amino acids Q, R, K, S or T.

SEQ ID NO: 39 (IhfA5-mIhfB4NTHI head chimeric peptide)

where "X" is an optional amino acid linker sequence, optionally comprising from 1 to 20 amino acids.

SEQ ID NO: 40 (IhfA5-mIhfB4NTHI head chimeric peptide)

SEQ ID NO: 41 (IhfA3-IhfB2NTHI tail chimeric peptide)

FLEEIRLSLESGQDVKLSGF-X- TLSAKEIENMVKDILEFISQ

SEQ ID NO. 42: Non-limiting exemplary linker: GGSGGS.

SEQ ID NO. 43: Non-limiting exemplary linker: GPSLKL.

SEQ ID NO. 44: Non-limiting exemplary linker: GGG.

SEQ ID NO. 45: Non-limiting exemplary linker: GPSL.

SEQ ID NO. 46: Non-limiting exemplary linker: GPS.

SEQ ID NO. 47: Non-limiting exemplary linker: PSLK.

SEQ ID NO. 48: Non-limiting exemplary linker: GPSLK.

SEQ ID NO. 49: non-limiting example of linker: SLKL.

SEQ ID NO: 50 (IhfA3-IhfB2NTHI tail chimeric peptide)

FLEIRLSLESGQDVKLSGFGPSLTLSAKEIENMVKDILEFISQ

--->

SEQUENCE LIST

<110> RESEARCH INSTITUTE AT NATIONWIDE CHILDREN'S HOSPITAL

<120> ANTIBODY COMPOSITIONS FOR DISRUPTING BIOFILMS

<130> 106887-0860

<140> PCT/US2020/041082

<141> 2020-07-07

<150> 63/033,109

<151> 2020-06-01

<150> 62/871,457

<151> 2019-07-08

<160> 50

<170> PatentIn version 3.5

<210> 1

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 1

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Lys Leu Val Glu Ser Gly

20 25 30

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

35 40 45

Ser Gly Phe Thr Phe Arg Thr Tyr Ala Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Gly Ser Asp Arg Arg

65 70 75 80

His Thr Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

85 90 95

Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Val Gly Pro Tyr Asp Gly Tyr Tyr

115 120 125

Gly Glu Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 2

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 2

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Val Glu Ser Gly

20 25 30

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

35 40 45

Ser Gly Phe Thr Phe Arg Thr Tyr Ala Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Gly Ser Asp Arg Arg

65 70 75 80

His Thr Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

85 90 95

Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Val Gly Pro Tyr Asp Gly Tyr Tyr

115 120 125

Gly Glu Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 3

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 3

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Lys Leu Val Gln Ser Gly

20 25 30

Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala

35 40 45

Ser Gly Phe Thr Phe Arg Thr Tyr Ala Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Gln Arg Leu Glu Trp Val Ala Thr Ile Gly Ser Asp Arg Arg

65 70 75 80

His Thr Tyr Tyr Pro Asp Lys Phe Gln Gly Arg Val Thr Ile Thr Arg

85 90 95

Asp Asn Ala Lys Asn Thr Leu Tyr Met Glu Leu Ser Ser Leu Arg Ser

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Val Gly Pro Tyr Asp Gly Tyr Tyr

115 120 125

Gly Glu Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 4

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 4

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Val Glu Ser Gly

20 25 30

Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

35 40 45

Ser Gly Phe Thr Phe Ser Arg Tyr Gly Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Ser Ser Gly Gly Ser

65 70 75 80

Tyr Thr Tyr Tyr Thr Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

85 90 95

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Glu Arg His Gly Gly Asp Gly Tyr

115 120 125

Trp Tyr Phe Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 5

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 5

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Val Glu Ser Gly

20 25 30

Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

35 40 45

Ser Gly Phe Thr Phe Ser Arg Tyr Gly Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Lys Gly Leu Glu Trp Val Ser Thr Ile Ser Ser Gly Gly Ser

65 70 75 80

Tyr Thr Tyr Tyr Thr Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

85 90 95

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Glu Arg His Gly Gly Asp Gly Tyr

115 120 125

Trp Tyr Phe Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 6

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 6

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Val Glu Ser Gly

20 25 30

Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Thr Ala

35 40 45

Ser Gly Phe Thr Phe Ser Arg Tyr Gly Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Ser Ser Gly Gly Ser

65 70 75 80

Tyr Thr Tyr Tyr Thr Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

85 90 95

Asp Asn Ala Lys Asn Ile Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Glu Arg His Gly Gly Asp Gly Tyr

115 120 125

Trp Tyr Phe Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 7

<211> 239

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 7

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro

20 25 30

Val Thr Leu Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser

35 40 45

Leu Leu Asp Ser Asp Gly Lys Thr Phe Leu Asn Trp Leu Gln Gln Arg

50 55 60

Pro Gly Gln Ser Pro Arg Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp

65 70 75 80

Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

85 90 95

Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr

100 105 110

Cys Trp Gln Gly Thr His Phe Pro Tyr Thr Phe Gly Gln Gly Thr Lys

115 120 125

Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro

130 135 140

Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu

145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp

165 170 175

Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp

180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys

195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln

210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 8

<211> 239

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 8

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro

20 25 30

Val Thr Leu Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser

35 40 45

Leu Leu Asp Ser Asp Gly Lys Thr Phe Leu Asn Trp Leu Gln Gln Arg

50 55 60

Pro Gly Gln Ser Pro Arg Arg Leu Ile Tyr Leu Val Ser Lys Arg Asp

65 70 75 80

Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

85 90 95

Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr

100 105 110

Cys Trp Gln Gly Thr His Phe Pro Tyr Thr Phe Gly Gln Gly Thr Lys

115 120 125

Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro

130 135 140

Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu

145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp

165 170 175

Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp

180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys

195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln

210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 9

<211> 239

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 9

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Val Val Met Thr Gln Ser Pro Asp Ser Leu Ala

20 25 30

Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Asp Ser Asp Gly Lys Thr Phe Leu Asn Trp Leu Gln Gln Lys

50 55 60

Pro Gly Gln Pro Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp

65 70 75 80

Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

85 90 95

Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr

100 105 110

Cys Trp Gln Gly Thr His Phe Pro Tyr Thr Phe Gly Gln Gly Thr Lys

115 120 125

Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro

130 135 140

Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu

145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp

165 170 175

Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp

180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys

195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln

210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 10

<211> 233

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 10

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

20 25 30

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp

35 40 45

Ile Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val

50 55 60

Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser

65 70 75 80

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser

85 90 95

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Gly Asn

100 105 110

Pro Leu Arg Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr

115 120 125

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

130 135 140

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

145 150 155 160

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

165 170 175

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

180 185 190

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

195 200 205

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

210 215 220

Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230

<210> 11

<211> 233

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 11

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

20 25 30

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp

35 40 45

Ile Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val

50 55 60

Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser

65 70 75 80

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser

85 90 95

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn

100 105 110

Pro Leu Arg Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr

115 120 125

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

130 135 140

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

145 150 155 160

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

165 170 175

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

180 185 190

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

195 200 205

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

210 215 220

Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230

<210> 12

<211> 233

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 12

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser

20 25 30

Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp

35 40 45

Ile Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Val

50 55 60

Arg Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Ile Pro Ala

65 70 75 80

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser

85 90 95

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Gly Asn

100 105 110

Pro Leu Arg Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr

115 120 125

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

130 135 140

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

145 150 155 160

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

165 170 175

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

180 185 190

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

195 200 205

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

210 215 220

Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230

<210> 13

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (27)..(27)

<223> Any amino acid, such as Q or K

<220>

<221> MOD_RES

<222> (30)..(30)

<223> Any amino acid, such as E or Q

<220>

<221> MOD_RES

<222> (33)..(33)

<223> Any amino acid, such as G or A

<220>

<221> MOD_RES

<222> (34)..(34)

<223> Any amino acid, such as G or E

<220>

<221> MOD_RES

<222> (35)..(35)

<223> Any amino acid, such as L or V

<220>

<221> MOD_RES

<222> (36)..(36)

<223> Any amino acid, such as V or K

<220>

<221> MOD_RES

<222> (37)..(37)

<223> Any amino acid, such as Q or K, or absent

<220>

<221> MOD_RES

<222> (40)..(40)

<223> Any amino acid, such as R, G or A

<220>

<221> MOD_RES

<222> (42)..(42)

<223> Any amino acid, such as L or V

<220>

<221> MOD_RES

<222> (43)..(43)

<223> Any amino acid, such as R or K

<220>

<221> MOD_RES

<222> (44)..(44)

<223> Any amino acid, such as L or V

<220>

<221> MOD_RES

<222> (47)..(47)

<223> Any amino acid, such as T, A or K

<220>

<221> MOD_RES

<222> (54)..(54)

<223> Any amino acid, such as S or R, or absent

<220>

<221> MOD_RES

<222> (55)..(55)

<223> Any amino acid, such as R or T, or absent

<220>

<221> MOD_RES

<222> (57)..(57)

<223> Any amino acid, such as G or A, or absent

<220>

<221> MOD_RES

<222> (67)..(67)

<223> Any amino acid, such as K or Q

<220>

<221> MOD_RES

<222> (68)..(68)

<223> Any amino acid, such as G or R

<220>

<221> MOD_RES

<222> (73)..(73)

<223> Any amino acid, such as A or S

<220>

<221> MOD_RES

<222> (76)..(76)

<223> Any amino acid, such as S or G, or absent

<220>

<221> MOD_RES

<222> (78)..(78)

<223> Any amino acid, such as G or D, or absent

<220>

<221> MOD_RES

<222> (79)..(79)

<223> Any amino acid, such as G or R, or absent

<220>

<221> MOD_RES

<222> (80)..(80)

<223> Any amino acid, such as S or R, or absent

<220>

<221> MOD_RES

<222> (81)..(81)

<223> Any amino acid, such as Y or H, or absent

<220>

<221> MOD_RES

<222> (85)..(85)

<223> Any amino acid, such as T or P, or absent

<220>

<221> MOD_RES

<222> (87)..(87)

<223> Any amino acid, such as S or K

<220>

<221> MOD_RES

<222> (88)..(88)

<223> Any amino acid, such as V or F

<220>

<221> MOD_RES

<222> (89)..(89)

<223> Any amino acid, such as K or Q

<220>

<221> MOD_RES

<222> (92)..(92)

<223> Any amino acid, such as F or V

<220>

<221> MOD_RES

<222> (95)..(95)

<223> Any amino acid, such as S or T

<220>

<221> MOD_RES

<222> (99)..(99)

<223> Any amino acid, such as A or S

<220>

<221> MOD_RES

<222> (102)..(102)

<223> Any amino acid, such as I, S or T

<220>

<221> MOD_RES

<222> (105)..(105)

<223> Any amino acid, such as L or M

<220>

<221> MOD_RES

<222> (106)..(106)

<223> Any amino acid, such as Q or E

<220>

<221> MOD_RES

<222> (107)..(107)

<223> Any amino acid, such as M or L

<220>

<221> MOD_RES

<222> (108)..(108)

<223> Any amino acid, such as N or S

<220>

<221> MOD_RES

<222> (111)..(111)

<223> Any amino acid, such as K or R

<220>

<221> MOD_RES

<222> (112)..(112)

<223> Any amino acid, such as T, A or S

<220>

<221> MOD_RES

<222> (121)..(121)

<223> Any amino acid, such as E or V, or absent

<220>

<221> MOD_RES

<222> (122)..(122)

<223> Any amino acid, such as R or G, or absent

<220>

<221> MOD_RES

<222> (123)..(123)

<223> Any amino acid, such as H or P, or absent

<220>

<221> MOD_RES

<222> (124)..(124)

<223> Any amino acid, such as G or Y, or absent

<220>

<221> MOD_RES

<222> (125)..(125)

<223> Any amino acid, such as G or D, or absent

<220>

<221> MOD_RES

<222> (126)..(126)

<223> Any amino acid, such as D or G, or absent

<220>

<221> MOD_RES

<222> (127)..(127)

<223> Any amino acid, such as G or Y, or absent

<220>

<221> MOD_RES

<222> (129)..(129)

<223> Any amino acid, such as W or G, or absent

<220>

<221> MOD_RES

<222> (130)..(130)

<223> Any amino acid, such as Y or E, or absent

<220>

<221> MOD_RES

<222> (133)..(133)

<223> Any amino acid, such as V or Y, or absent

<220>

<221> MOD_RES

<222> (139)..(139)

<223> Any amino acid, such as M or L, or absent

<400> 13

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Xaa Leu Val Xaa Ser Gly

20 25 30

Xaa Xaa Xaa Xaa Xaa Pro Gly Xaa Ser Xaa Xaa Xaa Ser Cys Xaa Ala

35 40 45

Ser Gly Phe Thr Phe Xaa Xaa Tyr Xaa Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Xaa Xaa Leu Glu Trp Val Xaa Thr Ile Xaa Ser Xaa Xaa Xaa

65 70 75 80

Xaa Thr Tyr Tyr Xaa Asp Xaa Xaa Xaa Gly Arg Xaa Thr Ile Xaa Arg

85 90 95

Asp Asn Xaa Lys Asn Xaa Leu Tyr Xaa Xaa Xaa Xaa Ser Leu Xaa Xaa

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr

115 120 125

Xaa Xaa Phe Asp Xaa Trp Gly Gln Gly Thr Xaa Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 14

<211> 239

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (22)..(22)

<223> Any amino acid, such as I or V, or absent

<220>

<221> MOD_RES

<222> (23)..(23)

<223> Any amino acid, such as V or Q

<220>

<221> MOD_RES

<222> (29)..(29)

<223> Any amino acid, such as A, S, D or L, or absent

<220>

<221> MOD_RES

<222> (30)..(30)

<223> Any amino acid, such as T or S

<220>

<221> MOD_RES

<222> (32)..(32)

<223> Any amino acid, such as S, A or P

<220>

<221> MOD_RES

<222> (33)..(33)

<223> Any amino acid, such as L, A or V

<220>

<221> MOD_RES

<222> (34)..(34)

<223> Any amino acid, such as S or T

<220>

<221> MOD_RES

<222> (35)..(35)

<223> Any amino acid, such as P, V or L

<220>

<221> MOD_RES

<222> (37)..(37)

<223> Any amino acid, such as E, D or Q, or absent

<220>

<221> MOD_RES

<222> (38)..(38)

<223> Any amino acid, such as R or P

<220>

<221> MOD_RES

<222> (39)..(39)

<223> Any amino acid, such as A or V

<220>

<221> MOD_RES

<222> (40)..(40)

<223> Any amino acid, such as T or S

<220>

<221> MOD_RES

<222> (41)..(41)

<223> Any amino acid, such as L or I

<220>

<221> MOD_RES

<222> (42)..(42)

<223> Any amino acid, such as S, T or N

<220>

<221> MOD_RES

<222> (44)..(44)

<223> Any amino acid, such as R or K

<220>

<221> MOD_RES

<222> (45)..(45)

<223> Any amino acid, such as A or S, or absent

<220>

<221> MOD_RES

<222> (48)..(48)

<223> Any amino acid, such as D or S, or absent

<220>

<221> MOD_RES

<222> (49)..(49)

<223> Any amino acid, such as I or L, or absent

<220>

<221> MOD_RES

<222> (50)..(50)

<223> Any amino acid, such as L or absent

<220>

<221> MOD_RES

<222> (51)..(51)

<223> Any amino acid, such as D or absent

<220>

<221> MOD_RES

<222> (52)..(52)

<223> Any amino acid, such as S or absent

<220>

<221> MOD_RES

<222> (53)..(53)

<223> Any amino acid, such as D or absent

<220>

<221> MOD_RES

<222> (54)..(54)

<223> Any amino acid, such as G or absent

<220>

<221> MOD_RES

<222> (55)..(55)

<223> Any amino acid, such as S or K, or absent

<220>

<221> MOD_RES

<222> (56)..(56)

<223> Any amino acid, such as N or T, or absent

<220>

<221> MOD_RES

<222> (57)..(57)

<223> Any amino acid, such as Y or F, or absent

<220>

<221> MOD_RES

<222> (61)..(61)

<223> Any amino acid, such as Y or L, or absent

<220>

<221> MOD_RES

<222> (64)..(64)

<223> Any amino acid, such as K or R

<220>

<221> MOD_RES

<222> (67)..(67)

<223> Any amino acid, such as Q or K

<220>

<221> MOD_RES

<222> (68)..(68)

<223> Any amino acid, such as A, P or S

<220>

<221> MOD_RES

<222> (69)..(69)

<223> Any amino acid, such as V or P, or absent

<220>

<221> MOD_RES

<222> (70)..(70)

<223> Any amino acid, such as R or K, or absent

<220>

<221> MOD_RES

<222> (71)..(71)

<223> Any amino acid, such as L or R, or absent

<220>

<221> MOD_RES

<222> (75)..(75)

<223> Any amino acid, such as Y or L, or absent

<220>

<221> MOD_RES

<222> (76)..(76)

<223> Any amino acid, such as T or V, or absent

<220>

<221> MOD_RES

<222> (78)..(78)

<223> Any amino acid, such as R or K, or absent

<220>

<221> MOD_RES

<222> (79)..(79)

<223> Any amino acid, such as L or R

<220>

<221> MOD_RES

<222> (80)..(80)

<223> Any amino acid, such as H or D, or absent

<220>

<221> MOD_RES

<222> (83)..(83)

<223> Any amino acid, such as I or V

<220>

<221> MOD_RES

<222> (85)..(85)

<223> Any amino acid, such as A, S or D

<220>

<221> MOD_RES

<222> (96)..(96)

<223> Any amino acid, such as Y or F, or absent

<220>

<221> MOD_RES

<222> (99)..(99)

<223> Any amino acid, such as T or K

<220>

<221> MOD_RES

<222> (102)..(102)

<223> Any amino acid, such as S or R

<220>

<221> MOD_RES

<222> (103)..(103)

<223> Any amino acid, such as L or V

<220>

<221> MOD_RES

<222> (104)..(104)

<223> Any amino acid, such as E or Q, or absent

<220>

<221> MOD_RES

<222> (105)..(105)

<223> Any amino acid, such as P or A, or absent

<220>

<221> MOD_RES

<222> (108)..(108)

<223> Any amino acid, such as F or V, or absent

<220>

<221> MOD_RES

<222> (109)..(109)

<223> Any amino acid, such as A or G

<220>

<221> MOD_RES

<222> (110)..(110)

<223> Any amino acid, such as V or T

<220>

<221> MOD_RES

<222> (112)..(112)

<223> Any amino acid, such as F or Y

<220>

<221> MOD_RES

<222> (114)..(114)

<223> Any amino acid, such as Q or W, or absent

<220>

<221> MOD_RES

<222> (117)..(117)

<223> Any amino acid, such as N or T, or absent

<220>

<221> MOD_RES

<222> (118)..(118)

<223> Any amino acid, such as P or H, or absent

<220>

<221> MOD_RES

<222> (119)..(119)

<223> Any amino acid, such as F or absent

<220>

<221> MOD_RES

<222> (120)..(120)

<223> Any amino acid, such as L or P, or absent

<220>

<221> MOD_RES

<222> (121)..(121)

<223> Any amino acid, such as R or Y, or absent

<220>

<221> MOD_RES

<222> (125)..(125)

<223> Any amino acid, such as G or Q, or absent

<220>

<221> MOD_RES

<222> (129)..(129)

<223> Any amino acid, such as V or L, or absent

<400> 14

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Xaa Xaa Met Thr Gln Ser Pro Xaa Xaa Leu Xaa

20 25 30

Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Cys Xaa Xaa Ser Gln Xaa

35 40 45

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Asn Trp Xaa Gln Gln Xaa

50 55 60

Pro Gly Xaa Xaa Xaa Xaa Xaa Leu Ile Tyr Xaa Xaa Ser Xaa Xaa Xaa

65 70 75 80

Ser Gly Xaa Pro Xaa Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Xaa

85 90 95

Thr Leu Xaa Ile Ser Xaa Xaa Xaa Xaa Glu Asp Xaa Xaa Xaa Tyr Xaa

100 105 110

Cys Xaa Gln Gly Xaa Xaa Xaa Xaa Xaa Thr Phe Gly Xaa Gly Thr Lys

115 120 125

Xaa Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro

130 135 140

Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu

145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp

165 170 175

Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp

180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys

195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln

210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 15

<211> 384

<212> PRT

<213> Homo sapiens

<400> 15

Ala Pro Thr Lys Ala Pro Asp Val Phe Pro Ile Ile Ser Gly Cys Arg

1 5 10 15

His Pro Lys Asp Asn Ser Pro Val Val Leu Ala Cys Leu Ile Thr Gly

20 25 30

Tyr His Pro Thr Ser Val Thr Val Thr Trp Tyr Met Gly Thr Gln Ser

35 40 45

Gln Pro Gln Arg Thr Phe Pro Glu Ile Gln Arg Arg Asp Ser Tyr Tyr

50 55 60

Met Thr Ser Ser Gln Leu Ser Thr Pro Leu Gln Gln Trp Arg Gln Gly

65 70 75 80

Glu Tyr Lys Cys Val Val Gln His Thr Ala Ser Lys Ser Lys Lys Glu

85 90 95

Ile Phe Arg Trp Pro Glu Ser Pro Lys Ala Gln Ala Ser Ser Val Pro

100 105 110

Thr Ala Gln Pro Gln Ala Glu Gly Ser Leu Ala Lys Ala Thr Thr Ala

115 120 125

Pro Ala Thr Thr Arg Asn Thr Gly Arg Gly Gly Glu Glu Lys Lys Lys

130 135 140

Glu Lys Glu Lys Glu Glu Gln Glu Glu Arg Glu Thr Lys Thr Pro Glu

145 150 155 160

Cys Pro Ser His Thr Gln Pro Leu Gly Val Tyr Leu Leu Thr Pro Ala

165 170 175

Val Gln Asp Leu Trp Leu Arg Asp Lys Ala Thr Phe Thr Cys Phe Val

180 185 190

Val Gly Ser Asp Leu Lys Asp Ala His Leu Thr Trp Glu Val Ala Gly

195 200 205

Lys Val Pro Thr Gly Gly Val Glu Glu Gly Leu Leu Glu Arg His Ser

210 215 220

Asn Gly Ser Gln Ser Gln His Ser Arg Leu Thr Leu Pro Arg Ser Leu

225 230 235 240

Trp Asn Ala Gly Thr Ser Val Thr Cys Thr Leu Asn His Pro Ser Leu

245 250 255

Pro Pro Gln Arg Leu Met Ala Leu Arg Glu Pro Ala Ala Gln Ala Pro

260 265 270

Val Lys Leu Ser Leu Asn Leu Leu Ala Ser Ser Asp Pro Pro Glu Ala

275 280 285

Ala Ser Trp Leu Leu Cys Glu Val Ser Gly Phe Ser Pro Pro Asn Ile

290 295 300

Leu Leu Met Trp Leu Glu Asp Gln Arg Glu Val Asn Thr Ser Gly Phe

305 310 315 320

Ala Pro Ala Arg Pro Pro Pro Gln Pro Gly Ser Thr Thr Phe Trp Ala

325 330 335

Trp Ser Val Leu Arg Val Pro Ala Pro Pro Ser Pro Gln Pro Ala Thr

340 345 350

Tyr Thr Cys Val Val Ser His Glu Asp Ser Arg Thr Leu Leu Asn Ala

355 360 365

Ser Arg Ser Leu Glu Val Ser Tyr Val Thr Asp His Gly Pro Met Lys

370 375 380

<210> 16

<211> 330

<212> PRT

<213> Homo sapiens

<400> 16

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 17

<211> 326

<212> PRT

<213> Homo sapiens

<400> 17

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

100 105 110

Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

115 120 125

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

130 135 140

Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly

145 150 155 160

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn

165 170 175

Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp

180 185 190

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro

195 200 205

Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu

210 215 220

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn

225 230 235 240

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

245 250 255

Ser Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

260 265 270

Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

275 280 285

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

290 295 300

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

305 310 315 320

Ser Leu Ser Pro Gly Lys

325

<210> 18

<211> 377

<212> PRT

<213> Homo sapiens

<400> 18

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro

100 105 110

Arg Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg

115 120 125

Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys

130 135 140

Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys Pro

145 150 155 160

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

165 170 175

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

180 185 190

Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Lys Trp Tyr

195 200 205

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

210 215 220

Gln Tyr Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Leu His

225 230 235 240

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

245 250 255

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln

260 265 270

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met

275 280 285

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

290 295 300

Ser Asp Ile Ala Val Glu Trp Glu Ser Ser Gly Gln Pro Glu Asn Asn

305 310 315 320

Tyr Asn Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu

325 330 335

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Ile

340 345 350

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn Arg Phe Thr Gln

355 360 365

Lys Ser Leu Ser Leu Ser Pro Gly Lys

370 375

<210> 19

<211> 452

<212> PRT

<213> Homo sapiens

<400> 19

Gly Ser Ala Ser Ala Pro Thr Leu Phe Pro Leu Val Ser Cys Glu Asn

1 5 10 15

Ser Pro Ser Asp Thr Ser Ser Val Ala Val Gly Cys Leu Ala Gln Asp

20 25 30

Phe Leu Pro Asp Ser Ile Thr Leu Ser Trp Lys Tyr Lys Asn Asn Ser

35 40 45

Asp Ile Ser Ser Thr Arg Gly Phe Pro Ser Val Leu Arg Gly Gly Lys

50 55 60

Tyr Ala Ala Thr Ser Gln Val Leu Leu Pro Ser Lys Asp Val Met Gln

65 70 75 80

Gly Thr Asp Glu His Val Val Cys Lys Val Gln His Pro Asn Gly Asn

85 90 95

Lys Glu Lys Asn Val Pro Leu Pro Val Ile Ala Glu Leu Pro Pro Lys

100 105 110

Val Ser Val Phe Val Pro Pro Arg Asp Gly Phe Phe Gly Asn Pro Arg

115 120 125

Lys Ser Lys Leu Ile Cys Gln Ala Thr Gly Phe Ser Pro Arg Gln Ile

130 135 140

Gln Val Ser Trp Leu Arg Glu Gly Lys Gln Val Gly Ser Gly Val Thr

145 150 155 160

Thr Asp Gln Val Gln Ala Glu Ala Lys Glu Ser Gly Pro Thr Thr Tyr

165 170 175

Lys Val Thr Ser Thr Leu Thr Ile Lys Glu Ser Asp Trp Leu Gly Gln

180 185 190

Ser Met Phe Thr Cys Arg Val Asp His Arg Gly Leu Thr Phe Gln Gln

195 200 205

Asn Ala Ser Ser Met Cys Val Pro Asp Gln Asp Thr Ala Ile Arg Val

210 215 220

Phe Ala Ile Pro Pro Ser Phe Ala Ser Ile Phe Leu Thr Lys Ser Thr

225 230 235 240

Lys Leu Thr Cys Leu Val Thr Asp Leu Thr Thr Tyr Asp Ser Val Thr

245 250 255

Ile Ser Trp Thr Arg Gln Asn Gly Glu Ala Val Lys Thr His Thr Asn

260 265 270

Ile Ser Glu Ser His Pro Asn Ala Thr Phe Ser Ala Val Gly Glu Ala

275 280 285

Ser Ile Cys Glu Asp Asp Trp Asn Ser Gly Glu Arg Phe Thr Cys Thr

290 295 300

Val Thr His Thr Asp Leu Pro Ser Pro Leu Lys Gln Thr Ile Ser Arg

305 310 315 320

Pro Lys Gly Val Ala Leu His Arg Pro Asp Val Tyr Leu Leu Pro Pro

325 330 335

Ala Arg Glu Gln Leu Asn Leu Arg Glu Ser Ala Thr Ile Thr Cys Leu

340 345 350

Val Thr Gly Phe Ser Pro Ala Asp Val Phe Val Gln Trp Met Gln Arg

355 360 365

Gly Gln Pro Leu Ser Pro Glu Lys Tyr Val Thr Ser Ala Pro Met Pro

370 375 380

Glu Pro Gln Ala Pro Gly Arg Tyr Phe Ala His Ser Ile Leu Thr Val

385 390 395 400

Ser Glu Glu Glu Trp Asn Thr Gly Glu Thr Tyr Thr Cys Val Ala His

405 410 415

Glu Ala Leu Pro Asn Arg Val Thr Glu Arg Thr Val Asp Lys Ser Thr

420 425 430

Gly Lys Pro Thr Leu Tyr Asn Val Ser Leu Val Met Ser Asp Thr Ala

435 440 445

Gly Thr Cys Tyr

450

<210> 20

<211> 327

<212> PRT

<213> Homo sapiens

<400> 20

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly Lys

325

<210> 21

<211> 353

<212> PRT

<213> Homo sapiens

<400> 21

Ala Ser Pro Thr Ser Pro Lys Val Phe Pro Leu Ser Leu Cys Ser Thr

1 5 10 15

Gln Pro Asp Gly Asn Val Val Ile Ala Cys Leu Val Gln Gly Phe Phe

20 25 30

Pro Gln Glu Pro Leu Ser Val Thr Trp Ser Glu Ser Gly Gln Gly Val

35 40 45

Thr Ala Arg Asn Phe Pro Pro Ser Gln Asp Ala Ser Gly Asp Leu Tyr

50 55 60

Thr Thr Ser Ser Gln Leu Thr Leu Pro Ala Thr Gln Cys Leu Ala Gly

65 70 75 80

Lys Ser Val Thr Cys His Val Lys His Tyr Thr Asn Pro Ser Gln Asp

85 90 95

Val Thr Val Pro Cys Pro Val Pro Ser Thr Pro Pro Thr Pro Ser Pro

100 105 110

Ser Thr Pro Pro Thr Pro Ser Pro Ser Cys Cys His Pro Arg Leu Ser

115 120 125

Leu His Arg Pro Ala Leu Glu Asp Leu Leu Leu Gly Ser Glu Ala Asn

130 135 140

Leu Thr Cys Thr Leu Thr Gly Leu Arg Asp Ala Ser Gly Val Thr Phe

145 150 155 160

Thr Trp Thr Pro Ser Ser Gly Lys Ser Ala Val Gln Gly Pro Pro Glu

165 170 175

Arg Asp Leu Cys Gly Cys Tyr Ser Val Ser Ser Val Leu Pro Gly Cys

180 185 190

Ala Glu Pro Trp Asn His Gly Lys Thr Phe Thr Cys Thr Ala Ala Tyr

195 200 205

Pro Glu Ser Lys Thr Pro Leu Thr Ala Thr Leu Ser Lys Ser Gly Asn

210 215 220

Thr Phe Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser Glu Glu Leu

225 230 235 240

Ala Leu Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg Gly Phe Ser

245 250 255

Pro Lys Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln Glu Leu Pro

260 265 270

Arg Glu Lys Tyr Leu Thr Trp Ala Ser Arg Gln Glu Pro Ser Gln Gly

275 280 285

Thr Thr Thr Phe Ala Val Thr Ser Ile Leu Arg Val Ala Ala Glu Asp

290 295 300

Trp Lys Lys Gly Asp Thr Phe Ser Cys Met Val Gly His Glu Ala Leu

305 310 315 320

Pro Leu Ala Phe Thr Gln Lys Thr Ile Asp Arg Leu Ala Gly Lys Pro

325 330 335

Thr His Val Asn Val Ser Val Val Met Ala Glu Val Asp Gly Thr Cys

340 345 350

Tyr

<210> 22

<211> 340

<212> PRT

<213> Homo sapiens

<400> 22

Ala Ser Pro Thr Ser Pro Lys Val Phe Pro Leu Ser Leu Asp Ser Thr

1 5 10 15

Pro Gln Asp Gly Asn Val Val Val Ala Cys Leu Val Gln Gly Phe Phe

20 25 30

Pro Gln Glu Pro Leu Ser Val Thr Trp Ser Glu Ser Gly Gln Asn Val

35 40 45

Thr Ala Arg Asn Phe Pro Pro Ser Gln Asp Ala Ser Gly Asp Leu Tyr

50 55 60

Thr Thr Ser Ser Gln Leu Thr Leu Pro Ala Thr Gln Cys Pro Asp Gly

65 70 75 80

Lys Ser Val Thr Cys His Val Lys His Tyr Thr Asn Pro Ser Gln Asp

85 90 95

Val Thr Val Pro Cys Pro Val Pro Pro Pro Pro Pro Cys Cys His Pro

100 105 110

Arg Leu Ser Leu His Arg Pro Ala Leu Glu Asp Leu Leu Leu Gly Ser

115 120 125

Glu Ala Asn Leu Thr Cys Thr Leu Thr Gly Leu Arg Asp Ala Ser Gly

130 135 140

Ala Thr Phe Thr Trp Thr Pro Ser Ser Gly Lys Ser Ala Val Gln Gly

145 150 155 160

Pro Pro Glu Arg Asp Leu Cys Gly Cys Tyr Ser Val Ser Ser Val Leu

165 170 175

Pro Gly Cys Ala Gln Pro Trp Asn His Gly Glu Thr Phe Thr Cys Thr

180 185 190

Ala Ala His Pro Glu Leu Lys Thr Pro Leu Thr Ala Asn Ile Thr Lys

195 200 205

Ser Gly Asn Thr Phe Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser

210 215 220

Glu Glu Leu Ala Leu Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg

225 230 235 240

Gly Phe Ser Pro Lys Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln

245 250 255

Glu Leu Pro Arg Glu Lys Tyr Leu Thr Trp Ala Ser Arg Gln Glu Pro

260 265 270

Ser Gln Gly Thr Thr Thr Phe Ala Val Thr Ser Ile Leu Arg Val Ala

275 280 285

Ala Glu Asp Trp Lys Lys Gly Asp Thr Phe Ser Cys Met Val Gly His

290 295 300

Glu Ala Leu Pro Leu Ala Phe Thr Gln Lys Thr Ile Asp Arg Met Ala

305 310 315 320

Gly Lys Pro Thr His Val Asn Val Ser Val Val Met Ala Glu Val Asp

325 330 335

Gly Thr Cys Tyr

340

<210> 23

<211> 106

<212> PRT

<213> Homo sapiens

<400> 23

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

1 5 10 15

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

20 25 30

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

35 40 45

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

50 55 60

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

65 70 75 80

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

85 90 95

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 24

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (27)..(27)

<223> K or Q

<220>

<221> MOD_RES

<222> (30)..(30)

<223> Q or E

<220>

<221> MOD_RES

<222> (33)..(33)

<223> A or G

<220>

<221> MOD_RES

<222> (34)..(34)

<223> E or G

<220>

<221> MOD_RES

<222> (35)..(35)

<223> V or L

<220>

<221> MOD_RES

<222> (36)..(36)

<223> K or V

<220>

<221> MOD_RES

<222> (37)..(37)

<223> K or Q

<220>

<221> MOD_RES

<222> (40)..(40)

<223> A or G

<220>

<221> MOD_RES

<222> (42)..(42)

<223> V or L

<220>

<221> MOD_RES

<222> (43)..(43)

<223> K or R

<220>

<221> MOD_RES

<222> (44)..(44)

<223> V or L

<220>

<221> MOD_RES

<222> (47)..(47)

<223> K or A

<220>

<221> MOD_RES

<222> (67)..(67)

<223> Q or K

<220>

<221> MOD_RES

<222> (68)..(68)

<223> R or G

<220>

<221> MOD_RES

<222> (87)..(87)

<223> K or S

<220>

<221> MOD_RES

<222> (88)..(88)

<223> F or V

<220>

<221> MOD_RES

<222> (89)..(89)

<223> Q or K

<220>

<221> MOD_RES

<222> (92)..(92)

<223> V or F

<220>

<221> MOD_RES

<222> (95)..(95)

<223> T or S

<220>

<221> MOD_RES

<222> (99)..(99)

<223> A or S

<220>

<221> MOD_RES

<222> (105)..(105)

<223> M or L

<220>

<221> MOD_RES

<222> (106)..(106)

<223> E or Q

<220>

<221> MOD_RES

<222> (107)..(107)

<223> L or M

<220>

<221> MOD_RES

<222> (108)..(108)

<223> S or N

<220>

<221> MOD_RES

<222> (112)..(112)

<223> S or A

<400> 24

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Xaa Leu Val Xaa Ser Gly

20 25 30

Xaa Xaa Xaa Xaa Xaa Pro Gly Xaa Ser Xaa Xaa Xaa Ser Cys Xaa Ala

35 40 45

Ser Gly Phe Thr Phe Arg Thr Tyr Ala Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Xaa Xaa Leu Glu Trp Val Ala Thr Ile Gly Ser Asp Arg Arg

65 70 75 80

His Thr Tyr Tyr Pro Asp Xaa Xaa Xaa Gly Arg Xaa Thr Ile Xaa Arg

85 90 95

Asp Asn Xaa Lys Asn Thr Leu Tyr Xaa Xaa Xaa Xaa Ser Leu Arg Xaa

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Val Gly Pro Tyr Asp Gly Tyr Tyr

115 120 125

Gly Glu Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 25

<211> 239

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (29)..(29)

<223> D or L

<220>

<221> MOD_RES

<222> (32)..(32)

<223> A or P

<220>

<221> MOD_RES

<222> (34)..(34)

<223> S or T

<220>

<221> MOD_RES

<222> (37)..(37)

<223> E or Q

<220>

<221> MOD_RES

<222> (38)..(38)

<223> R or P

<220>

<221> MOD_RES

<222> (40)..(40)

<223> T or S

<220>

<221> MOD_RES

<222> (42)..(42)

<223> N or S

<220>

<221> MOD_RES

<222> (44)..(44)

<223> K or R

<220>

<221> MOD_RES

<222> (64)..(64)

<223> K or R

<220>

<221> MOD_RES

<222> (68)..(68)

<223> P or S

<220>

<221> MOD_RES

<222> (70)..(70)

<223> K or R

<220>

<221> MOD_RES

<222> (79)..(79)

<223> L or R

<220>

<221> MOD_RES

<222> (99)..(99)

<223> T or K

<220>

<221> MOD_RES

<222> (102)..(102)

<223> S or R

<220>

<221> MOD_RES

<222> (103)..(103)

<223> L or V

<220>

<221> MOD_RES

<222> (104)..(104)

<223> Q or E

<220>

<221> MOD_RES

<222> (109)..(109)

<223> A or G

<400> 25

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Val Val Met Thr Gln Ser Pro Xaa Ser Leu Xaa

20 25 30

Val Xaa Leu Gly Xaa Xaa Ala Xaa Ile Xaa Cys Xaa Ser Ser Gln Ser

35 40 45

Leu Leu Asp Ser Asp Gly Lys Thr Phe Leu Asn Trp Leu Gln Gln Xaa

50 55 60

Pro Gly Gln Xaa Pro Xaa Arg Leu Ile Tyr Leu Val Ser Lys Xaa Asp

65 70 75 80

Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

85 90 95

Thr Leu Xaa Ile Ser Xaa Xaa Xaa Ala Glu Asp Val Xaa Val Tyr Tyr

100 105 110

Cys Trp Gln Gly Thr His Phe Pro Tyr Thr Phe Gly Gln Gly Thr Lys

115 120 125

Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro

130 135 140

Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu

145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp

165 170 175

Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp

180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys

195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln

210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 26

<211> 473

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (37)..(37)

<223> Q or K

<220>

<221> MOD_RES

<222> (40)..(40)

<223> R or G

<220>

<221> MOD_RES

<222> (47)..(47)

<223> T or A

<220>

<221> MOD_RES

<222> (73)..(73)

<223> A or S

<220>

<221> MOD_RES

<222> (102)..(102)

<223> I or S

<220>

<221> MOD_RES

<222> (111)..(111)

<223> K or R

<220>

<221> MOD_RES

<222> (112)..(112)

<223> T or A

<400> 26

Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser

1 5 10 15

Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Val Glu Ser Gly

20 25 30

Gly Gly Leu Val Xaa Pro Gly Xaa Ser Leu Arg Leu Ser Cys Xaa Ala

35 40 45

Ser Gly Phe Thr Phe Ser Arg Tyr Gly Met Ser Trp Val Arg Gln Ala

50 55 60

Pro Gly Lys Gly Leu Glu Trp Val Xaa Thr Ile Ser Ser Gly Gly Ser

65 70 75 80

Tyr Thr Tyr Tyr Thr Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

85 90 95

Asp Asn Ala Lys Asn Xaa Leu Tyr Leu Gln Met Asn Ser Leu Xaa Xaa

100 105 110

Glu Asp Thr Ala Val Tyr Tyr Cys Glu Arg His Gly Gly Asp Gly Tyr

115 120 125

Trp Tyr Phe Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

130 135 140

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

145 150 155 160

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

165 170 175

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

180 185 190

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

195 200 205

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

210 215 220

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

225 230 235 240

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

245 250 255

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

260 265 270

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

275 280 285

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

290 295 300

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

305 310 315 320

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

325 330 335

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

340 345 350

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

355 360 365

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

370 375 380

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

385 390 395 400

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

405 410 415

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

420 425 430

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

435 440 445

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

450 455 460

Gln Lys Ser Leu Ser Leu Ser Pro Gly

465 470

<210> 27

<211> 233

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (23)..(23)

<223> V or Q

<220>

<221> MOD_RES

<222> (29)..(29)

<223> A or S

<220>

<221> MOD_RES

<222> (30)..(30)

<223> T or S

<220>

<221> MOD_RES

<222> (33)..(33)

<223> L or A

<220>

<221> MOD_RES

<222> (35)..(35)

<223> P or V

<220>

<221> MOD_RES

<222> (37)..(37)

<223> E or D

<220>

<221> MOD_RES

<222> (39)..(39)

<223> A or V

<220>

<221> MOD_RES

<222> (41)..(41)

<223> L or I

<220>

<221> MOD_RES

<222> (42)..(42)

<223> S or T

<220>

<221> MOD_RES

<222> (62)..(62)

<223> Q or K

<220>

<221> MOD_RES

<222> (65)..(65)

<223> R or K

<220>

<221> MOD_RES

<222> (78)..(78)

<223> I or V

<220>

<221> MOD_RES

<222> (80)..(80)

<223> A or S

<220>

<221> MOD_RES

<222> (99)..(99)

<223> E or Q

<220>

<221> MOD_RES

<222> (105)..(105)

<223> V or T

<220>

<221> MOD_RES

<222> (107)..(107)

<223> F or Y

<400> 27

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Xaa Met Thr Gln Ser Pro Xaa Xaa Leu Ser

20 25 30

Xaa Ser Xaa Gly Xaa Arg Xaa Thr Xaa Xaa Cys Arg Ala Ser Gln Asp

35 40 45

Ile Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Xaa Ala Val

50 55 60

Xaa Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Xaa Pro Xaa

65 70 75 80

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser

85 90 95

Ser Leu Xaa Pro Glu Asp Phe Ala Xaa Tyr Xaa Cys Gln Gln Gly Asn

100 105 110

Pro Leu Arg Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr

115 120 125

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

130 135 140

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

145 150 155 160

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

165 170 175

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

180 185 190

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

195 200 205

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

210 215 220

Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230

<210> 28

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 28

Trp Ala Thr Cys Ala Ala Asn Asn Asn Asn Thr Thr Arg

1 5 10

<210> 29

<211> 90

<212> PRT

<213> Escherichia coli

<400> 29

Met Asn Lys Thr Gln Leu Ile Asp Val Ile Ala Glu Lys Ala Glu Leu

1 5 10 15

Ser Lys Thr Gln Ala Lys Ala Ala Leu Glu Ser Thr Leu Ala Ala Ile

20 25 30

Thr Glu Ser Leu Lys Glu Gly Asp Ala Val Gln Leu Val Gly Phe Gly

35 40 45

Thr Phe Lys Val Asn His Arg Ala Glu Arg Thr Gly Arg Asn Pro Gln

50 55 60

Thr Gly Lys Glu Ile Lys Ile Ala Ala Ala Asn Val Pro Ala Phe Val

65 70 75 80

Ser Gly Lys Ala Leu Lys Asp Ala Val Lys

85 90

<210> 30

<211> 90

<212> PRT

<213> Escherichia coli

<400> 30

Met Asn Lys Ser Gln Leu Ile Asp Lys Ile Ala Ala Gly Ala Asp Ile

1 5 10 15

Ser Lys Ala Ala Ala Gly Arg Ala Leu Asp Ala Ile Ile Ala Ser Val

20 25 30

Thr Glu Ser Leu Lys Glu Gly Asp Asp Val Ala Leu Val Gly Phe Gly

35 40 45

Thr Phe Ala Val Lys Glu Arg Ala Ala Arg Thr Gly Arg Asn Pro Gln

50 55 60

Thr Gly Lys Glu Ile Thr Ile Ala Ala Ala Lys Val Pro Ser Phe Arg

65 70 75 80

Ala Gly Lys Ala Leu Lys Asp Ala Val Asn

85 90

<210> 31

<211> 21

<212> PRT

<213> Haemophilus influenzae

<400> 31

Asn Phe Glu Leu Arg Asp Lys Ser Ser Arg Pro Gly Arg Asn Pro Lys

1 5 10 15

Thr Gly Asp Val Val

20

<210> 32

<211> 21

<212> PRT

<213> Haemophilus influenzae

<400> 32

Ser Leu His His Arg Gln Pro Arg Leu Gly Arg Asn Pro Lys Thr Gly

1 5 10 15

Asp Ser Val Asn Leu

20

<210> 33

<211> 6

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 33

Gly Pro Ser Leu Lys Leu

1 5

<210> 34

<211> 4

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 34

Gly Pro Ser Leu

1

<210> 35

<211> 4

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 35

Pro Ser Leu Lys

1

<210> 36

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 36

Gly Pro Ser Leu Lys

1 5

<210> 37

<211> 4

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 37

Ser Leu Lys Leu

1

<210> 38

<211> 60

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (7)..(7)

<223> Any amino acid

<220>

<221> MOD_RES

<222> (21)..(40)

<223> Any amino acid

<220>

<221> MISC_FEATURE

<222> (21)..(40)

<223> This region may encompass 1-20 residues

<220>

<221> MOD_RES

<222> (55)..(55)

<223> Any amino acid

<400> 38

Arg Pro Gly Arg Asn Pro Xaa Thr Gly Asp Val Val Pro Val Ser Ala

1 5 10 15

Arg Arg Val Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Ser Leu His His Arg Gln Pro

35 40 45

Arg Leu Gly Arg Asn Pro Xaa Thr Gly Asp Ser Val

50 55 60

<210> 39

<211> 60

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (21)..(40)

<223> Any amino acid

<220>

<221> MISC_FEATURE

<222> (21)..(40)

<223> This region may encompass 1-20 residues

<400> 39

Arg Pro Gly Arg Asn Pro Lys Thr Gly Asp Val Val Pro Val Ser Ala

1 5 10 15

Arg Arg Val Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Ser Leu His His Arg Gln Pro

35 40 45

Arg Leu Gly Arg Asn Pro Lys Thr Gly Asp Ser Val

50 55 60

<210> 40

<211> 44

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 40

Arg Pro Gly Arg Asn Pro Lys Thr Gly Asp Val Val Pro Val Ser Ala

1 5 10 15

Arg Arg Val Val Gly Pro Ser Leu Phe Ser Leu His His Arg Gln Pro

20 25 30

Arg Leu Gly Arg Asn Pro Lys Thr Gly Asp Ser Val

35 40

<210> 41

<211> 60

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<220>

<221> MOD_RES

<222> (21)..(40)

<223> Any amino acid

<220>

<221> MISC_FEATURE

<222> (21)..(40)

<223> This region may encompass 1-20 residues

<400> 41

Phe Leu Glu Glu Ile Arg Leu Ser Leu Glu Ser Gly Gln Asp Val Lys

1 5 10 15

Leu Ser Gly Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr Leu Ser Ala Lys Glu Ile Glu

35 40 45

Asn Met Val Lys Asp Ile Leu Glu Phe Ile Ser Gln

50 55 60

<210> 42

<211> 6

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 42

Gly Gly Ser Gly Gly Ser

1 5

<210> 43

<211> 6

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 43

Gly Pro Ser Leu Lys Leu

1 5

<210> 44

<211> 3

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 44

Gly Gly Gly

1

<210> 45

<211> 4

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 45

Gly Pro Ser Leu

1

<210> 46

<211> 3

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 46

Gly Pro Ser

1

<210> 47

<211> 4

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 47

Pro Ser Leu Lys

1

<210> 48

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 48

Gly Pro Ser Leu Lys

1 5

<210> 49

<211> 4

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

peptide

<400> 49

Ser Leu Lys Leu

1

<210> 50

<211> 44

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of Artificial Sequence: Synthetic

polypeptide

<400> 50

Phe Leu Glu Glu Ile Arg Leu Ser Leu Glu Ser Gly Gln Asp Val Lys

1 5 10 15

Leu Ser Gly Phe Gly Pro Ser Leu Thr Leu Ser Ala Lys Glu Ile Glu

20 25 30

Asn Met Val Lys Asp Ile Leu Glu Phe Ile Ser Gln

35 40

<---

Claims (40)

1. An antibody or antigen-binding fragment thereof that specifically binds to the head region of the DNABII peptide, comprising: (a) the variable domain of the heavy chain (HC) of an immunoglobulin, containing: (i) a heavy chain complementarity determining region 1 (CDRH1) comprising the amino acid sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1, or 2, or 3, or 24); (ii) a heavy chain complementarity determining region 2 (CDRH2) comprising the amino acid sequence GSDRRH (aa 76-81 of SEQ ID NO: 1, or 2, or 3, or 24); (iii) a heavy chain complementarity determining region 3 (CDRH3) comprising the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1, or 2, or 3, or 24), and (b) a variable domain of a light chain (LC) of an immunoglobulin comprising: (i) a complementarity determining region of the light chain 1 (CDRL1) comprising the amino acid sequence QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7, or 8, or 9, or 25); (ii) a light chain complementarity determining region 2 (CDRL2) comprising the amino acid sequence LVS (aa 75-77 of SEQ ID NO: 7, or 8, or 9, or 25); and (iii) a complementarity determining region of the light chain 3 (CDRL3) comprising the amino acid sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7, or 8, or 9, or 25). 2. An antibody or antigen-binding fragment thereof according to claim 1, characterized in that the sequence of the variable domain of the HC immunoglobulin contains an amino acid sequence that is at least 90% identical to aa 25-144 of SEQ ID NO: 1, and/or characterized in that the sequence of the variable domain of the LC immunoglobulin contains an amino acid sequence that is at least 90% identical to aa 21-132 of SEQ ID NO: 7. 3. An antibody or antigen-binding fragment thereof according to claim 1, characterized in that the sequence of the variable domain of the HC immunoglobulin contains an amino acid sequence that is at least 90% identical to aa 25-144 of SEQ ID NO: 1, and/or characterized in that the sequence of the variable domain of the LC immunoglobulin contains an amino acid sequence that is at least 90% identical to aa 21-132 of SEQ ID NO: 8. 4. An antibody or antigen-binding fragment thereof according to claim 1, characterized in that the sequence of the variable domain of the HC immunoglobulin contains an amino acid sequence that is at least 90% identical to aa 25-144 of SEQ ID NO: 1, and/or characterized in that the sequence of the variable domain of the LC immunoglobulin contains an amino acid sequence that is at least 90% identical to aa 21-132 of SEQ ID NO: 9. 5. An antibody or antigen-binding fragment thereof according to any one of claims 1-4, characterized in that the antibody also contains a constant region selected from the group consisting of: an IgA constant region, an IgD constant region, an IgE constant region, an IgG constant region, or an IgM constant region. 6. An antibody or antigen-binding fragment thereof according to claim 5, characterized in that the constant region is an IgG1 constant region. 7. An antibody or antigen-binding fragment thereof according to any one of claims 1-4, characterized in that the antigen-binding fragment is selected from the group consisting of Fab, F(ab')2, scFv or Fv. 8. A derivative of an antibody or antigen-binding fragment thereof that specifically binds to the head region of the DNABII peptide, comprising: (a) the variable domain of the heavy chain (HC) of an immunoglobulin, containing: (i) a heavy chain complementarity determining region 1 (CDRH1) comprising the amino acid sequence GFTFRTY (aa 50-56 of SEQ ID NO: 1, or 2, or 3, or 24); (ii) a heavy chain complementarity determining region 2 (CDRH2) comprising the amino acid sequence GSDRRH (aa 76-81 of SEQ ID NO: 1, or 2, or 3, or 24); (iii) a heavy chain complementarity determining region 3 (CDRH3) comprising the sequence VGPYDGYYGEFDY (aa 121-133 of SEQ ID NO: 1, or 2, or 3, or 24), and (b) a variable domain of a light chain (LC) of an immunoglobulin comprising: (i) a complementarity determining region of the light chain 1 (CDRL1) comprising the amino acid sequence QSLLDSDGKTF (aa 47-57 of SEQ ID NO: 7, or 8, or 9, or 25); (ii) a light chain complementarity determining region 2 (CDRL2) comprising the amino acid sequence LVS (aa 75-77 of SEQ ID NO: 7, or 8, or 9, or 25); and (iii) a complementarity determining region of the light chain 3 (CDRL3) comprising the amino acid sequence WQGTHFPYT (aa 114-122 of SEQ ID NO: 7, or 8, or 9, or 25), wherein one or more amino acids in the antibody or antigen-binding fragment thereof are chemically modified by alkylation, PEGylation, PEGmimetation, acylation, polysialylation, HESylation or glycosylation, ester formation or amide formation. 9. A derivative of an antibody or antigen-binding fragment thereof according to claim 8, wherein the derivative of the antibody or antigen-binding fragment thereof is modified, wherein the modification is selected from the group consisting of PEGylation, PEGmimetation, polysialylation, HESylation, or glycosylation. 10. An isolated polynucleotide encoding an antibody or antigen-binding fragment thereof according to any one of claims 1-7. 11. An expression vector containing the isolated polynucleotide according to claim 10. 12. The expression vector of claim 11, wherein the isolated polynucleotide is operably linked to a promoter and/or enhancer element. 13. A host cell for producing an antibody or an antigen-binding fragment thereof according to any one of claims 1-7, containing a polynucleotide according to claim 10 or an expression vector according to claim 11 or 12. 14. A composition for preventing the formation or destruction of a biofilm, comprising a carrier and one or more of: an antibody or antigen-binding fragment thereof according to any one of claims 1-7, a derivative of an antibody or antigen-binding fragment thereof according to claim 8 or 9, an isolated polynucleotide according to claim 10, an expression vector according to claim 11 or 12, or a host cell according to claim 13. 15. The composition according to claim 14, characterized in that the carrier comprises a component with a buffering capacity, including alanine, arginine, glycine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine or aspartame; and optionally the carrier comprises a carbohydrate excipient selected from monosaccharides, disaccharides, polysaccharides and/or alditols, wherein optionally the monosaccharides are selected from fructose, maltose, galactose, glucose, D-mannose and sorbose; optionally the disaccharides are selected from lactose, sucrose, trehalose and cellobiose; optionally the polysaccharides are selected from raffinose, melezitose, maltodextrins, dextrans and starches; and optionally the alditols are selected from mannitol, xylitol, maltitol, lactitol, xylitol, sorbitol (glucitol) and myoinositol. 16. A method for producing an antibody or an antigen-binding fragment thereof according to any one of claims 1-7, comprising culturing a host cell containing the isolated polynucleotide according to claim 10, under conditions that ensure expression of the polynucleotide. 17. The method of claim 16, further comprising isolating the antibody or antigen-binding fragment thereof. 18. A method for producing a derivative of an antibody or an antigen-binding fragment thereof according to any one of claims 8 or 9, comprising culturing a host cell containing the isolated polynucleotide of claim 10 under conditions that allow expression of the antibody or its antigen-binding fragment, and subsequently modifying the antibody or its antigen-binding fragment, such as post-translational modification or covalent attachment of the molecule. 19. A method for preventing the formation or destruction of a biofilm on a surface, comprising treating a surface susceptible to a biofilm or containing a biofilm with an antibody or an antigen-binding fragment thereof according to any one of claims 1-7 or a derivative of an antibody or an antigen-binding fragment thereof according to claim 8 or 9. 20. A method for preventing the formation or destruction of a biofilm in a subject, comprising administering to the subject an antibody or antigen-binding fragment thereof according to any one of claims 1-7 or a derivative of an antibody or antigen-binding fragment thereof according to claim 8 or 9. 21. A method for preventing or treating a microbial infection that causes biofilm formation in a subject, comprising administering to the subject an antibody or antigen-binding fragment thereof according to any one of claims 1-7 or an antibody derivative or antigen-binding fragment thereof according to claim 8 or 9. 22. A method of treating a condition characterized by biofilm formation in a subject, comprising administering to the subject an antibody or antigen-binding fragment thereof according to any one of claims 1-7 or an antibody derivative or antigen-binding fragment thereof according to claim 8 or 9. 23. Use of an antibody or antigen-binding fragment according to any one of claims 1-7 or a derivative of an antibody or antigen-binding fragment thereof according to any one of claims 8, 9 for coating a non-physiological surface, optionally where the surface is under industrial conditions.