[go: up one dir, main page]

WO2010036031A2 - Anticorps monoclonal humain specifique au pauf, composition pharmaceutique destinee au traitement du cancer le comprenant et procede l'utilisant pour detecter le cancer - Google Patents

Anticorps monoclonal humain specifique au pauf, composition pharmaceutique destinee au traitement du cancer le comprenant et procede l'utilisant pour detecter le cancer Download PDF

Info

Publication number
WO2010036031A2
WO2010036031A2 PCT/KR2009/005437 KR2009005437W WO2010036031A2 WO 2010036031 A2 WO2010036031 A2 WO 2010036031A2 KR 2009005437 W KR2009005437 W KR 2009005437W WO 2010036031 A2 WO2010036031 A2 WO 2010036031A2
Authority
WO
WIPO (PCT)
Prior art keywords
cancer
pauf
seq
protein
antibody
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2009/005437
Other languages
English (en)
Other versions
WO2010036031A3 (fr
Inventor
Sang Seok Koh
Yang Soon Lee
Mi Jin Sohn
Su Jin Kim
Kyung Sook Yoo
Eun Hye Park
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Korea Research Institute of Bioscience and Biotechnology KRIBB
Original Assignee
Korea Research Institute of Bioscience and Biotechnology KRIBB
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Korea Research Institute of Bioscience and Biotechnology KRIBB filed Critical Korea Research Institute of Bioscience and Biotechnology KRIBB
Publication of WO2010036031A2 publication Critical patent/WO2010036031A2/fr
Publication of WO2010036031A3 publication Critical patent/WO2010036031A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/303—Liver or Pancreas
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation

Definitions

  • the present invention relates to a PAUF protein associated with the onset of cancer, a human monoclonal antibody specific to the PAUF protein, and a nucleic acid molecule coding for the PAUF protein, and uses thereof. More particularly, the present invention relates to a human monoclonal antibody binding specifically to a PAUF protein, a diagnostic marker for cancer comprising a PAUF protein or PAUF nucleic acid, a di- agnositic composition for cancer comprising an antibody specific to the marker, a method for the diagnosis of cancer using the composition, a method of screening a material therapeutic or preventive of cancer by determining the expression level of the marker, and a composition for the treatment or prevention of cancer featuring the inhibitory activity against the marker.
  • pancreatic cancer accounts for about 2% of all instances of cancer. In spite of the low rate of incidence, pancreatic cancer has one of the highest mortality rates of all cancers and is fourth in the list of cancers which result in death in Korea. Patients diagnosed with pancreatic cancer typically have a poor prognosis partly because the cancer usually exhibits no symptoms early on, leading to locally advanced or metastatic disease at the time of diagnosis. In many cases, pancreatic cancer has developed into the final stages and has spread over too wide of a region for surgical resection to be viable. Importantly, there are no effective curatives (Faint et al. (2004) Datamonitor DMHC2045; Garcea et al. (2005) Pancreatology 5:514-529).
  • pancreatic cancer there have been no methods used to diagnose pancreatic cancer from human bodily fluids including blood.
  • MRI or CT is able to diagnose pancreatic cancer, but only after it enters the later stage.
  • CA19-9 assay is too low in specificity to be applied to the diagnosis of pancreatic cancer, and is used only for monitoring the treatment of diseases (Notification of Ministry for health, welfare and family affairs: Method for diagnosis of Cancer).
  • the only chance patients with pancreatic cancer have for a cure is complete surgical resection of the tumor.
  • 10 ⁇ 15% of progressive pancreatic cancer cases are typically found to be potentially operable due to the presence of distant metastasis.
  • chemotherapeutic agents in clinical use may be divided into chemotherapeutic agents and biotherapeutic agents.
  • Chemotherapy in its most general sense, refers to treatment of disease by chemicals that kill cancer cells. Virtually all chemotherapeutic regimens can cause significant side effects such as inducing cytotoxicity in normal cells and inducing chemotherapy resistance as a result of the defense mechanism of tumor cells.
  • biotherapeutic agents have recently been actively developed because they can inhibit the progression of cancer by enhancing the immune system or by suppressing the activity of cancer cells.
  • chemotherapeutic agents examples include cytokines, recombinant antibodies such as monoclonal antibodies, gene therapy agents, and an- giogenesis inhibitors (Kim, Yeol Hong et al., (2004), Study on Cancer Gene and De- velopment of Novel Anticancer Agents, Overview of Health Industry Technology 2003 Summer 10-18).
  • Therapeutic monoclonal antibodies act only on disease-specific targets thanks to the high reaction specificity thereof and thus are known to show much lower side effects than do conventional cytotoxic chemicals.
  • the therapeutic effects of antibodies are obtained through various mechanisms.
  • antibodies may bind specifically to corresponding antigens to inhibit signaling or may induce apoptosis by cross-linking.
  • the therapeutic effects may be extracted by activating the immune system with antibodies.
  • anticancer agents monoclonal antibodies can be responsible for the functions of tracing cancer cells, suppressing the functions of targets, eliciting immune responses and effectively eliminating cancer cells. With these advantages, monoclonal antibodies are playing an increasingly major role in cancer therapy. 20 kinds of monoclonal antibodies were already approved for use in diagnosis and treatment in 2005. More than half of them were applied to the diagnosis and treatment of various cancers.
  • composition for diagnosing cancer comprising at least one selected from a group consisting of a protein, a peptide, an aptamer and an antibody, all of which specifically bind to a PAUF protein or an immunogenic fragment thereof.
  • PAUF protein comprising bringing the antibody into contact with a biological specimen to examine whether the level of PAUF protein is increased or decreased.
  • the present invention pertains to a human monoclonal antibody binding specifically to PAUF or a functional fragment thereof.
  • PAUF is overexpressed in cancer, particularly in pancreatic cancer, stomach cancer, ovarian cancer and colorectal cancer.
  • the present inventors found that PAUF is over- expressed in pancreatic cancer at the early stage, compared to normal cells. Moreover, overexpression of PAUF protein is observed in patients who have pancreatic cancer. In addition, it was observed that the growth, metastasis and invasion of cancer are likely to occur in cell lines expressing PAUF.
  • the present invention provides a human monoclonal antibody or functional fragment thereof specially binding to PAUF (Pancreatic adenocarcinoma upregulating factor), comprising a light chain variable region of which an amino acid sequence is selected from the group consisting of: (a) a light chain variable region comprising CDRl as defined by SEQ ID NO.25, CDR2 as defined by SEQ ID NO.
  • PAUF Pancreatic adenocarcinoma upregulating factor
  • the present invention provides a human monoclonal antibody or functional fragment thereof specially binding to PAUF (Pancreatic adenocarcinoma upregulating factor) comprising a heavy chain variable region of which an amino acid sequence is selected from the group consisting of: (a) a heavy chain variable region comprising CDRl as defined by SEQ ID NO.28, CDR2 as defined by SEQ ID NO.29, and CDR3 as defined by SEQ ID NO.30; (b) a heavy chain variable region comprising CDRl as defined by SEQ ID NO.34, CDR2 as defined by SEQ ID NO.35, and CDR3 as defined by SEQ ID NO.36; (c) a heavy chain variable region comprising CDRl as defined by SEQ ID NO.40, CDR2 as defined by SEQ ID NO.41, and CDR3 as defined by SEQ ID NO.42; and (d) a heavy chain variable region comprising CDRl as defined by SEQ ID NO.46, CDR2 as defined by SEQ ID NO.
  • 'PAUF Pancreatic Adenocarcinoma Upregulating Factor
  • PAUF Pancreatic Adenocarcinoma Upregulating Factor
  • the 'protein' or 'polypeptide' refers, in part, to a protein that has the human amino acid sequence depicted in SEQ ID NO. 50.
  • the terms also comprise naturally occurring allelic variants and proteins that have a slightly different amino acid sequence than that specifically recited above. Allelic variants, though possessing a slightly different amino acid sequence than those recited above, will still have the same or similar biological functions associated with these proteins.
  • SEQ ID NO. 50 refers to proteins that have been isolated from organisms in addition to humans.
  • PAUF protein is preferably in isolated form.
  • a protein is said to be isolated when physical, mechanical or chemical methods are employed to remove the protein from cellular constituents that are normally associated with the protein. A skilled artisan can readily employ standard purification methods to obtain an isolated protein.
  • the above PAUF protein further includes insertion, deletion or conservative amino acid substitution variants of SEQ ID NO. 50.
  • a conservative variant refers to alterations in the amino acid sequence that do not adversely affect the biological functions of the protein.
  • a substitution, insertion or deletion is said to adversely affect the protein when the altered sequence prevents or disrupts a biological function associated with the protein.
  • the overall charge, structure or hy- drophobic/hydrophilic properties of the protein in certain instances, may be altered without adversely affecting a biological activity.
  • the amino acid sequence can be altered, for example to render the peptide more hydrophobic or hydrophilic, without adversely affecting the biological activities of the protein.
  • allelic variants, the conservative substitution variants, and the members of the protein family will have an amino acid sequence having at least about 50%, 60%, 70% or 75% amino acid sequence identity with the sequence set forth in SEQ ID NO. 50, more preferably at least about 80-90%, even more preferably at least about 92-94%, and most preferably at least about 95%, 98% or 99% sequence identity.
  • Identity or homology with respect to such sequences is defined herein as the percentage of amino acid residues in the candidate sequence that are identical with SEQ ID NO. 50, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology, and not considering any conservative sub- stitutions as part of the sequence identity (see section B for the relevant parameters). Fusion proteins, or N-terminal, C-terminal or internal extensions, deletions, or insertions into the peptide sequence shall not be construed as affecting homology.
  • the above PAUF protein includes molecules having the amino acid sequence disclosed in SEQ ID NO. 50; amino acid sequence variants wherein one or more amino acid residues has been inserted N- or C-terminal to, or within, the disclosed coding sequence; and amino acid sequence variants of the disclosed sequence, or their fragments as defined above, that have been substituted by at least one residue.
  • Such fragments also referred to as peptides or polypeptides, may contain antigenic regions, functional regions of the protein identified as regions of the amino acid sequence which correspond to known protein domains, as well as regions of pronounced hy- drophilicity. The regions are all easily identifiable by using commonly available protein sequence analysis software such as Mac Vector (Oxford Molecular).
  • Contemplated variants further include those containing predetermined mutations by, e.g., homologous recombination, site-directed or PCR mutagenesis, and the corresponding proteins of other animal species, including but not limited to rabbit, mouse, rat, porcine, bovine, ovine, equine and non-human primate species, and the alleles or other naturally occurring variants of the family of proteins; and derivatives wherein the protein has been covalently modified by substitution, chemical, enzymatic, or other appropriate means with a moiety other than a naturally occurring amino acid (for example a detectable moiety such as an enzyme or radioisotope).
  • a detectable moiety such as an enzyme or radioisotope
  • the PAUF nucleic acid molecule has a sequence as defined by SEQ ID NO. 49.
  • the antibody specially binding to PAUF in accordance with the present invention comprises specific CDRs (Complementarity Determining Region) for binding to an antigen, but is not limited thereto.
  • the antibody specific for PAUF protein in accordance with the present invention may comprise CDRs (Complementarity Determining Region) which includes a specific sequence. More preferably, the monoclonal antibody comprises a CDR in the light chain or a CDR in the heavy chain, respectively, or all CDRs in the light and heavy chains. It will be understood by persons skilled in the art that the monoclonal antibody of the present invention may be generated by grafting CDRs onto framework regions (FR) of a known therapeutic antibody.
  • CDRs Complementarity Determining Region
  • the antibody specific to PAUF in accordance with the present invention may be polyclonal or monoclonal with preference for monoclonal antibody. Most preferably, the antibody is a human monoclonal antibody or a functional fragment thereof as described above.
  • the term 'monoclonal antibody' is recognized in the art and refers to a highly specific antibody directed by a single antigenic determinant (epitope).
  • polyclonal antibodies are a mixture of immunoglobulin molecules secreted against a specific antigen by different B cell lines, each recognizing a different epitope.
  • monoclonal antibodies are derived from a single cell line.
  • a polyclonal antibody is lower in specificity than a monoclonal antibody because it has cross- reactivity to different proteins.
  • antisera are different in antibody titer and specificity according to lots. For these reasons, drugs utilizing polyclonal antibodies are difficult to control in quality upon production.
  • monoclonal antibodies can improve diagnostic assay which is based on antigen- antibody binding by improving selectivity and specificity. Also advantageously, monoclonal antibodies are not contaminated with other immunoglobulins because they are produced by culturing hybridoma.
  • Antibodies to PAUF may be produced using typical methods known in the art, for example, the fusion method (Kohler and Milstein, European Journal of Immunology, 6:511-519 (1976)), the recombinant DNA method (U. S. Patent No. 4,816,56) or the phage antibody library method (Clackson et al, Nature, 352:624-628(1991) and Marks et al, J. MoI. Biol., 222:58, 1-597(1991)).
  • the fusion method Kelham and Milstein, European Journal of Immunology, 6:511-519 (1976)
  • the recombinant DNA method U. S. Patent No. 4,816,56
  • the phage antibody library method Clackson et al, Nature, 352:624-628(1991) and Marks et al, J. MoI. Biol., 222:58, 1-597(1991).
  • hybridoma cells producing monoclonal antibodies may be obtained by fusing immortal cell lines to antibody- producing lymphocytes the technologies for which are well known to those skilled in the art and can be easily carried out.
  • Polyclonal antibodies may be obtained by injecting a PAFU protein antigen into an appropriate animal, collecting antisera from the animal, and isolating antibodies from the antisera by an affinity technique.
  • human monoclonal antibodies specific to PAUF can be selected using well-known combinatorial antibody library and phage display technologies (see Example 1).
  • an antibody display library is constructed by cloning antibody genes, whether naive, immunized, or semi-synthesized, into a phagemid vector at the 5 'end site of gill and introducing the phagemids into E. coli.
  • the phagemid clones with antibody genes of interest inserted thereinto express antibody -pill fusion proteins in the host cells.
  • All of the structural proteins necessary for the assembly of recombinant phage virions, such as wild-type (wt) pill, etc., are supplied by helper phages such as M13K07 or VCSM13. Through a process called phage rescue, recombinant phage virions are produced.
  • recombinant phages display wt pill and antibody -pill fusion proteins at the same time on their surface.
  • the phagemid DNA is preferentially packaged inside the recombinant phage, resulting in the formation of genotype- phenotype linkage. Theoretically, this phage packaging process enforces oligovalent display of antibody fragments on the surface of the recombinant phage particles.
  • the phage display technology allows the separation of target molecule- specific antibody clones from the background of target molecule-non-specific clones by panning.
  • the human monoclonal antibody of the present invention can be used without purification or with purification by, for example, affinity chromatography, size-exclusion chromatography, ion-exchange chromatography, etc.
  • cell lines which can stably produce the antibodies according to the present invention are prepared.
  • the light chains and the heavy chains of the antibodies are inserted into an expression vector which is in turn transformed into a DHFR-deficient CHO cell line (CHO-DG44).
  • the transformed cells are cultured in selection media containing MTX (methotrexate), a DHFR inhibitor, at increasing concentrations.
  • MTX metalhotrexate
  • a DHFR inhibitor a DHFR inhibitor
  • any antibody may be used in the present invention.
  • the term 'functional fragment of an antibody' refers to a fragment which retains at least an antigen-binding function, and may include Fab, F(ab'), F(ab') 2 , and Fv.
  • the antibody according to the present invention or a functional fragment thereof, which can bind specifically to PAUF comprises (a) a light chain variable region defined as a polypeptide represented by one selected from a group consisting of amino acid sequences of SEQ ID Nos. 1 to 4, and/or (b) a heavy chain variable region defined as a polypeptide represented by one selected from a group consisting of amino acid sequences of SEQ ID Nos. 5 to 8.
  • the human monoclonal antibody according to the present invention comprises light chain amino acid sequence as defined by one selected from among SEQ ID NOS. 9 to 12; or/and a heavy chain amino acid sequence as defined by one selectd from among SEQ ID NOS. 13 to 16.
  • the present invention provides a nucleic acid molecule encoding the PAUF- specific monoclonal antibody or its fragment.
  • the nucleic acid molecule of the present invention comprises a base sequence selected from among base sequences of SEQ ID Nos. 17 to 20 coding respectively for the amino acid sequences of SEQ ID Nos. 1 to 4 for the light chain variable region, and/or a base sequence selected from among base sequences of SEQ ID Nos. 21 to 24 coding respectively for the amino acid sequences of SEQ ID Nos. 5 to 8 for the heavy chain variable region.
  • the human monoclonal antibody or its functional fragment in accordance with the present invention specifically binds to PAUF in various cancer cells to suppress the growth, migration and/or metastasis of cancer, leading to the death of tumor cells.
  • the cancer is pancreatic cancer, stomach cancer, ovarian cancer or colorectal cancer.
  • the present invention pertains to novel uses of PAUF protein or PAUF nucleic acid molecule as diagnostic markers or therapeutic targets in relation with cancer, based on the finding that PAUF is overexpressed in cancer and the growth, metastasis and invasion of cancer cells are likely to occur in the cell line where PAUF is over- expressed.
  • the present invention provides a di- agnostic marker for cancer, comprising a PAUF protein or a PAUF nucleic acid molecule coding for the same.
  • the diagnostic marker according to the present invention is selective for pancreatic cancer, stomach cancer and colorectal cancer and particularly specific for pancreatic cancer.
  • PAUF protein was detected in patients with pancreatic cancer, with a total diagnosis specificity of 100%.
  • pancreatic cancer specimens eight were decided to be positive, with a selectivity of 61.5%, and the other five negative specimens were observed to have slightly higher PAUF levels compared to normal specimens (FIG. 7), indicating that the PAUF protein can be useful as a diagnostic marker for cancer (see Example 8).
  • the present invention provides a diagnostic composition and kit for cancer, comprising at least one selected from a group consisting of a protein, a peptide, an aptamer and an antibody all of which bind specifically to a PAUF protein or an immunogenic fragment thereof.
  • the antibody is the human monoclonal antibody according to the present invention or a functional fragment thereof.
  • the present invention provides a method for detecting a PAUF protein using the human monoclonal antibody or the functional fragment thereof.
  • the detection of PAUF protein can be accomplished by bringing the human monoclonal antibody of the present invention or the functional fragment thereof into contact with a biological specimen to examine whether the level of PAUF protein increases or decreases.
  • the human monoclonal antibody or its functional fragment in accordance with the present invention specifically binds to PAUF in various cancer cells where PAUF protein is expressed, to suppress the growth, migration and/or metastasis of cancer, leading to the death of tumor cells.
  • the cancer is pancreatic cancer, stomach cancer, ovarian cancer or colorectal cancer.
  • composition or the kit in accordance with the present invention may further comprise another ingredient.
  • the composition or kit may further include a reagent capable of detecting an antigen- antibody complex, e.g., a labeled secondary antibody, a chromophore, an enzyme (i.e., conjugated with an antibody) and its substrate or other substances capable of binding to the antibody (FIG. 7).
  • the composition or kit may be in the form of multiple separate packages or compartments.
  • the diagnostic composition and kit in accordance with the present invention can be used to analyze the onset, progression and alleviation of cancer. Therefore, the term 'diagnosis', as used herein, is intended to include the meaning of determining the onset, progression and alleviation of diseases as well as determining the presence of diseases.
  • composition and kit of the present invention is useful in the diagnosis of the cancer whose onset and progression are associated with the overexpression of PAUF, that is, pancreatic cancer, stomach cancer, ovarian cancer and colorectal cancer, with preference for pancreatic cancer.
  • the diagnostic composition of the present invention is used to diagnose the conditions of cancer with high efficiency. Most advantageously, the diagnostic composition is applicable to the diagnosis of cancer when it is in its early stages.
  • the present invention provides a method of screening a material therapeutic or preventive of cancer, comprising:
  • test sample (c) judging the test sample as being therapeutic or preventive of cancer when the level or the activity is determined to be down-regulated.
  • the term 'prevention of cancer' is intended to refer to any action resulting in the suppression of carcinogenesis or the delay of the growth of cancer through the administration of a material obtained by the screening method of the present invention, or a composition comprising a material suppressive of the expression of PAUF protein or PAUF nucleic acid molecule or the activity of PAUF protein, for example, at least one selected from a group consisting of a protein, an aptamer, a peptide and an antibody, all of which bind specifically to PAUF.
  • treatment' is intended to refer to any action resulting in an improvement in the symptoms of the disease or which advantageously alters a disease state after a material has been administered, the material having been obtained by the screening method of the present invention or is composed of the composition thereof.
  • a test material to be analyzed is brought into contact with a cell containing a PAUF protein or a PAUF nucleic acid molecule.
  • test material is intended to mean an unknown material which is used in performing a screening method which is carried out for purposes of examining whether it has influence on the expression level of PAUF protein or PAUF nucleic acid molecule or the activity of PAUF protein or not.
  • test material useful in the present invention include chemicals, nucleotides, antisense-RNA, siRNA (small interferenceRNA), peptides, proteins, aptamers, and natural extracts, but are not limited thereto.
  • PAUF protein or PAUF nucleic acid molecule or the activity of PAUF protein Down- regulation in the level of PAUF protein or PAUF nucleic acid molecule or the activity of PAUF protein leads to the judgement that the test sample is therapeutic or preventive of cancer.
  • the term 'measurement of the level of protein or nucleic acid' is intended to refer to a process of examining whether and how much of the PAUF protein or the PAUF nucleic acid molecule is expressed in a biological sample.
  • the measurement of the level of PAUF nucleic acid molecule can be carried out using various well-known methods. For example, RT-PCR (Sambrook et al., Molecular Cloning. A Laboratory Manual, 3 rd ed. Cold Spring Harbor Press(2001)), Nothern blotting (Peter B. Kaufma et al., Molecular and Cellular Methods in Biology and Medicine, 102-108, CRCpress), hybridization on cDNA microarray (Sambrook et al., Molecular Cloning. A Laboratory Manual, 3 rd ed. Cold Spring Harbor Press (2001)) or in situ hybridization (Sambrook et al., Molecular Cloning. A Laboratory Manual, 3 rd ed. Cold Spring Harbor Press (2001)) may be used for the measurement.
  • RT-PCR Standardbrook et al., Molecular Cloning. A Laboratory Manual, 3 rd ed. Cold Spring Harbor Press(2001)
  • Nothern blotting Peter B
  • RNA is isolated from the treated cells, and a single-stranded cDNA is prepared with oligo dT primers in the presence of reverse transcriptase. PCR is performed using a gene-specific primer set with the cDNA serving as a template. After the electrophoresis of the PCR product, the bands thus formed are analyzed to determine the levels of PAUF nucleic acids in the cells.
  • PAUF protein As for the measurement of the level of PAUF protein, it can be obtained using various immunoassays known in the art. For example, a change in PAUF protein level can be detected using radioimmunoassay, radioimmunoprecipitation assay, immuno- precipitation, ELISA (enzyme-linked immunosorbentassay), capture-ELISA, suppressive or competitive ELISA, or sandwich ELISA.
  • radioimmunoassay radioimmunoassay
  • radioimmunoprecipitation assay immuno- precipitation
  • ELISA enzyme-linked immunosorbentassay
  • capture-ELISA capture-ELISA
  • suppressive or competitive ELISA or sandwich ELISA.
  • the protein level may be determined using a protein, an aptamer or a peptide, all of which bind specifically to PAUF protein, with preference for an antibody to PAUF protein.
  • Methods of measuring protein levels using antibodies include Western blotting,
  • ELISA enzyme linked immuNosorbent assay
  • RIA Radioimmunoassay
  • radioim- munodiffusion Ouchterlony immunodiffusion
  • rocket immunoelectrophoresis im- munohistostaining assay
  • immunoprecipitation assay complement fixation assay
  • FACS FACS, and protein chips, but is not limited thereto.
  • the term 'antigen-antibody complex' is intended to refer to binding products of the PAUF protein to an antibody specific thereto.
  • the antigen- antibody complex thus formed may be quantitatively determined by measuring the size of the signal generated by the detection label.
  • Such a detection label may be selected from a group consisting of enzymes, fluorescent substances, ligands, luminescent substances, microparticles, redox molecules and radioactive isotopes, but the present invention is not limited to these examples.
  • the present invention provides a pharmaceutical composition for the inhibition of cancer growth and metastasis, comprising as an active ingredient a material regulative of the expression of PAUF protein or PAUF nucleic acid molecule or the activity of PAUF protein.
  • the present invention provides a pharmaceutical composition for the treatment or prevention of cancer, comprising as an active ingredient a material regulative of the expression of PAUF protein or PAUF nucleic acid molecule or the activity of PAUF protein.
  • composition at least one selected from a group consisting of a protein, an aptamer, a peptide and an antibody, all of which bind specifically to PAUF, may be contained.
  • the pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier suitable for oral administration include a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, dye and a fragrant.
  • a buffer, a preservative, a pain reducer, a solubilizer, an isotonic agent, and a stabilizer may be used in combination.
  • the composition may further comprise a base, an excipient, a lubricant and/or a preservative.
  • the pharmaceutical composition of the present invention may be formulated, along with a pharmaceutically acceptable carrier, into various dosage forms.
  • the composition may be in the oral dosage form of tablets, troches, capsules, elixirs, suspensions, syrups or wafers.
  • the composition may be formulated into unit dose ampule forms or multiple dose ampule forms.
  • the composition may be applied to solutions, suspensions, tablets, pills, capsules, and suspended release forms.
  • Examples of carriers, excipients or diluents suitable for the formulation of the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, methyl cellulose, amorphous cellulose, polypyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.
  • a filler, an anticoagulant, a lubricant, a wetting agent, a fragrant and/or a preservative may be additionally used.
  • the effective dosage of the pharmaceutical composition in accordance with the present invention depends on various factors, including kinds of cancer, route of administration, patient's age, gender and weight, and severity of diseases, etc.
  • the composition of the present invention may further comprise a therapeutic agent which can be conjugated with the human monoclonal antibody of the present invention.
  • This therapeutic agent is selected from among ra- dionuclides, drugs, lymphokines, toxins, and bispecific antibodies.
  • the drugs or toxins include Etoposide, teniposide, adriamycin, daunomycin, carminomycin, aminopterin, dactinomycin, mitomycin, cis-platinum and cis-platinum analogues, bleomycins, esperamicins, 5-fluorouracil, melphalan, and nitrogen mustard, but are not limited thereto.
  • the PAUF-specific human monoclonal antibody or its functional fragment is used to inhibit the functions of PAUF in various cancers to suppress the growth and metastasis of the cancers, thereby killing the tumor cells.
  • the treatment or prevention of cancer is achieved by regulating the proliferation, migration or invasion of cancer.
  • the cancer is pancreatic cancer, stomach cancer, ovarian cancer, or colorectal cancer.
  • the method of the present invention comprises administering the pharmaceutical composition to the body in a pharmaceutically effective dosage.
  • the pharmaceutical composition may be administered parenterally, subcutaneously, intra- pulmonarily or intranasally.
  • the composition may, if desired, be administered using a suitable method including intralesional administration.
  • Parenteral injections include intramuscular, intravenous, intraarterial, intraperitoneal and subcutaneous routes.
  • Preferred administration modes include intravenous, subcutaneous, intradermal, intramuscular and drip injections.
  • the pancreatic cancer cell line Panc-1 was treated with the monoclonal antibody of the present invention and examined for proliferation to determine the effect of the monoclonal antibody on the growth of cancer cells.
  • the cells were found to proliferate at lower rates upon treatment with the antibody than with a control antibody (FIG. 3).
  • the monoclonal antibody according to the present invention inhibited the growth and/or survival of cancer cells.
  • the effect of the monoclonal antibody of the present invention on the migration of cancer cells was also examined.
  • the pancreatic cell lines CFPAC-I and Panc-1 were assayed for migration ability.
  • the cells treated with the antibody of the present invention were observed to decrease in migration ability as compared to those treated with a control antibody (FIG. 4).
  • the cancer cell lines treated with the antibody of the present invention were found to decease in invasion ability, compared to those treated with a control antibody, as measured by invasion assay (FIG. 5).
  • the monoclonal antibody of the present invention was examined for in vivo therapeutic activity against caner.
  • the pancreatic cancer cell line CFPAC-I was transplanted into immunodeficient mice.
  • the antibody was injected into the mice via the tail vein.
  • the mice treated with the antibody of the present invention were observed to decrease in tumor size (FIG. 6). Accordingly, the antibody of the present invention functioned to inhibit the progression of tumors in vivo.
  • Sequence lists are given in Table, below. They include amino acid sequences and corresponding base sequences for light chain variable regions and heavy chain variable regions of four anti-PAUF antibodies according to the present invention.
  • the PAUF protein and PAUF nucleic acid molecule according to the present invention is closely correlated with the onset of cancer and can be a marker or a therapeutic target particularly for pancreatic cancer.
  • the anti-PAUF antibody or its functional fragment according to the present invention binds specifically to PAUF on various cancer cells including pancreatic cancer and stomach cancer, to allow the diagnosis of cancer when it is in the early stages, to inhibit the growth, invasion or migration of cancer cells, and to induce the cancer cells to undergo apoptosis, so that it is useful in the diagnosis and treatment of cancer.
  • FIG. 1 shows purified monoclonal antibodies 8F3, 3A4, 36C9 and 6C4 on acrylamide gel.
  • FIG. 2 is a titer curve showing the binding of the antibodies 8F3, 3A4, 36C9 and 6C4 to PAUF protein.
  • FIG. 3 shows the inhibitory activity of the antibodies 8F3, 3A4, 36C9 and 6C4 against the growth of a pancreatic cancer cell line (Panc-1).
  • FIG. 4 shows the inhibitory activity of the antibodies 8F3, 3A4, 36C9 and 6C4 against the migration of two pancreatic cancer cell lines (Panc-1 and CFPAC-I).
  • FIG. 5 shows the inhibitory activity of the antibodies 8F3, 3A4, 36C9 and 6C4 against the invasion of two pancreatic cancer cell lines (Panc-1 and CFPAC-I).
  • FIG. 6 shows the inhibitory activity of the antibodies 8F3 and 36C9 against the growth of the pancreatic cancer cell line transplanted into mice.
  • FIG. 7 shows the ability of the humanized antibody 8F3 and polyclonal antibodies to PAUF to detect PAUF in blood and the detection of PAUF with the antibody specifically in sera from pancreatic cancer patients.
  • the PAUF protein was immobilized onto a plastic tube and buffered for 2 hours in a buffer (2% skim milk, PBS). After the removal of the buffer, a human antibody display phage library in a buffer was added to the plastic tube and incubated at room temperature for 2 hours. The tube was washed 10 times with a washing buffer (0.1% Tween 20, PBS) and then 10 times with PBS. Thereafter, the phages were eluted by incubation at room temperature for 10 min with 10OmM TEA (triethylamine). To the phage eluate was added IM Tris (pH 7.5), followed by incubation at room temperature for 5 min for neutralization. Then, an E.
  • a washing buffer (0.1% Tween 20, PBS
  • 10OmM TEA triethylamine
  • E. coli culture was added to the phage elutate.
  • the E. coli thus infected with the phage was recovered as a cell pellet by centrifugation at 3,000 xg for 5 min and spread over 2xYT plates.
  • E. coli colonies formed after incubation at 37 0 C for 16 hours were suspended in 2xYT media and cultured.
  • the infected E. coli after being recovered from the suspension, was inoculated into fresh 2xYT media and supplemented with helper phage to primarily screen the phage. With this primarily screened phage library, this screening process was further conducted four times. After five rounds of the screening, clones were randomly selected from 2xYT plates and cultured in 2xYT media.
  • helper phage To the clone culture was added helper phage to rescue phages.
  • the phages thus produced were examined for specificity for PAUF by PAUF ELISA (Example 3).
  • the phages which were found to show PAUF specificity were selected and subjected to base sequencing. Through these processes, 8F3, 3A4, 36C9 and 6C4 antibodies, which bind specifically to PAUF were excavated.
  • CHO-DG44 cell lines which can express PAUF-specific human monoclonal antibodies in a large quantity were established.
  • light chain variable regions of the 10 monoclonal antibodies excavated by use of the method described in Example 1 were inserted into the BstXI site of plgGLD while the heavy chain variable regions were inserted into the Sfil site of plgGHD, so that they were converted to IgG forms.
  • plgGLD and plgGHD are expression vectors for antibody production which contain constant regions of light and heavy chains of IgG, respectively.
  • the expression plasmids for IgG were transfected into the DHFR-deficient CHO cell line CHO-DG44 with the aid of Lipofectamin Plus (Invitrogen, CA) according to the protocol provided by the manufacturer.
  • the stably transfected cells were selected in the presence of 550 ⁇ g/m-6 of G418 (Invitogen, CA) over 2 weeks.
  • the stable transfectants were adapted to MTX (Sigma, MO) while the concentration was increased from 10 nM at intervals of two weeks.
  • the monoclonal antibodies secreted to the media from the adapted cell lines were quantitatively analyzed using ELISA. High-expression cell lines were selected and applied to the mass production of the monoclonal antibodies.
  • the selected cell lines were grown in SF CHO Media (JBI, KR) in tissue culture flasks to express the antibodies.
  • the antibodies of the present invention may be purified using various methods including affinity chromatography and ion exchange chromatography. All of the four antibodies 8F3, 3A4, 36C9 and 6C4 shown in FIG. 5 were purified as follows.
  • the human monoclonal antibodies secreted into the SF CHO Media were first subjected to affinity chromatography using Protein A columns which can bind to Fc regions of antibodies.
  • a culture medium containing the expressed antibodies was forced to pass once through a 0.45 ⁇ m membrane in a filter and loaded on Protein A columns which were pre-equilibrated with 20 mM sodium phosphate buffer (pH 7.0).
  • a 20 mM sodium phosphate buffer (pH 7.0) was drop wise added to the Protein A column to wash off unbound proteins.
  • the monoclonal antibodies were eluted with 100 mM sodium citrate buffer (pH3).
  • the pH of the resulting eluted fractions was adjusted into 7 ⁇ 7.5 with 1 M Tris buffer (pH 9.0).
  • the eluted monoclonal antibody fractions were pooled and concentrated using a 3K MWCO spin column with a buffer exchange with 20 mM sodium phosphate (pH 7.0). This sample was loaded on a cation exchange chromatography column which was previously equilibrated with 20 mM sodium phosphate buffer (pH7.0).
  • FIG. 1 shows purified monoclonal antibodies 8F3, 3A4, 36C9 and 6C4 on acrylamide gel.
  • the PAUF protein was immobilized onto a plate and buffered for 2 hours in a buffer (2% skim milk, PBS). After the removal of the buffer, an human monoclonal antibody display phage eluate or purified human antibodies in buffer was added to the plate and incubated at 37 0 C for 1 hour. Unbound antibodies were washed off five times with a washing solution (0.1% Tween 20, PBS). As a secondary antibody, anti-M13-HRP and anti-human IgG (Fc specific)-HRP were used for the phage elution or the human antibodies, respectively. A dilution (1:5000) of a suitable secondary antibody in buffer was added to the plate and incubated at 37 0 C for 30 min.
  • FIG. 2 shows measurements of antigen-binding capacity of the purified antibodies 8F3, 3A4, 36C9 and 6C4 according to concentrations.
  • Pancreatic cancer cells were plated at a density of 5xlO 4 cells/m ⁇ in 24-well plates and mixed with 3 ⁇ g/m-6 of the monoclonal antibody. The cells were incubated in a 37 0 C CO 2 incubator while the medium was exchanged with a fresh one every two days. After incubation for 6 days, cells were recovered stained with 0.2% tryphan blue, and counted. Comparison was made between the numbers of living cells in antibody- treated wells and in a control, indicating that the four monoclonal antibodies 8F3, 3A4, 36C9 and 6C4 could inhibit the growth of pancreatic cancer cells (FIG. 3).
  • Pancreatic cells were isolated and washed twice with PBS. Various concentrations of the antibodies were added in such an amount as to adjust the number of cells to 1 x 10 6 cells/m#.
  • the lower chamber was filled with 30 j ⁇ of a DMEM medium supplemented with 10% FBS and covered with a 8 m pore size polycarbonate filter (25x80 mm, Neuroprobe). Then, the upper chamber was applied to the lower chamber and immobilized. A mixture of pancreatic cancer cells and antibody was added in an amount of 50 ⁇ Jl per well to the immobilized upper chamber and incubated for 20 hours in a 37 0 C, CO 2 incubator.
  • the cells which migrated to the lower chamber were fixed with 100% methanol and stained with a Giemsa solution. In at least three selected areas of a photograph taken, the stained cells were counted. This assay indicated that the four selected monoclonal antibodies 8F3, 3A4, 36C9 and 6C4 could inhibit the migration of pancreatic cancer cells (FIG. 4).
  • Pancreatic cells were isolated and washed twice with PBS. Various concentrations of the antibodies were added in such an amount as to adjust the number of cells to 1 x 10 6 cells/m-6.
  • the lower chamber was filled with 30 id of a DMEM medium supplemented with 10% FBS and covered with a matrigel-coated, 8 m pore size polycarbonate filter (25x80 mm, Neuroprobe). Then, the upper chamber was applied to the lower chamber and immobilized. A mixture of pancreatic cancer cells and antibody was added in an amount of 50 id per well to the immobilized upper chamber and incubated for 20 hours in a 37 0 C CO 2 incubator.
  • the cells which migrated to the lower chamber were fixed with 100% methanol and stained with a Giemsa solution. In at least three selected areas of a photograph taken, the stained cells were counted. This assay indicated that the four selected monoclonal antibodies 8F3, 3A4, 36C9 and 6C4 could inhibit the invasion of pancreatic cancer cells (FIG. 5).
  • EXAMPLE 7 Assay of PAUF-Specific Antibodies for Inhibitory Activity against Growth of Cancer Cell
  • CFPAC-I cells were transplanted in an amount of 5 x 10 6 cells into the hypodermis of 6 ⁇ 7-week-old Balb/c nu/nu nude mice. From 10 days after the transplantation, the monoclonal antibodies were injected at a concentration of 5 mg/kg twice a week for three weeks via the tail vein. Using a caliper, tumor volumes were measured twice a week. On the final day of the experiment, the nude mice were sacrificed with CO 2 , and tumors were separated and weighed. The tumors were fixed, embedded in paraffin blocks, and stained with rabbit anti-PAUF polyclonal antibodies.
  • mice injected with the PAUF-specific monoclonal antibodies were found to be lower in PAUF level than those of the mice injected with a control antibody, indicating that the two selected monoclonal antibodies 8F3 and 36C9 can inhibit the growth of the pancreatic cell line CFPAC-I in mice (FIG. 6).
  • EXAMPLE 8 Measurement of Blood Level of PAUF with Anti-PAUF Antibody and Diagnosis of Pancreatic Cancer [200]
  • the following experiment was conducted in order to determine whether pancreatic cancer can be diagnosed by detecting PAUF in blood.
  • the anti-PAUF antibody 8F3 was diluted to a concentration of 1-10 ⁇ g/ml in PBS (pH 7.2) and the dilution was plated in an amount of 100 /i2/well onto 96- well plates, followed by incubation of the sealed plates at 4 0 C for 18 hours to sufficiently fix the antibody to the plates. After removal of unfixed antibodies, PBS containing 1% bovine serum albumin was added in an amount of 300 /i6/well onto the plates and incubated at 37 0 C for 2 hours.
  • HRP-conjugated anti-rabbit antibody was 1/2000 diluted in PBS containing 1% bovine serum and the dilution was plated in an amount of 100 /i6/well onto the plates, followed by incubation at 37 0 C for 30 min. The plates were washed five times with PBS containing 0.05% Tween20. A substrate solution containing 100 ⁇ g/ml tetrametyl benzidine, 0.006% hydrogen peroxide and citric acid in PBS (pH4.5) was added in an amount of 100 /i2/well to the plates and allowed to develop a color for 30 min in a dark place. Color development was terminated by adding 50 j ⁇ of a reaction stopper (IN sulfuric acid) to each well.
  • a reaction stopper IN sulfuric acid
  • Absorbance at 450 was read in a 96- well plate reader (Molecular Devices), with a reference wavelength at 650 nm.
  • the colorant tetramethyl benzidine was degraded by the HRP conjugate with antibody to develop a color. Color development was thus analyzed by measuring the absorbance at 450 nm, to determine the presence of PAUF in the blood.
  • the specimens used in this experiment were as follows.
  • the PAUF protein or the PAUF nucleic acid molecule in accordance with the present invention can be used as a diagnostic marker or a therapeutic target.
  • the anti-PAUF antibody or its functional fragment allows the diagnosis of cancer in the early stages thereof and can inhibit the growth, invasion or migration of cancer cells by inducing the cells to undergo apoptosis, and thus are useful in the diagnosis and treatment of cancer.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Cell Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne une protéine PAUF associée à la formation du cancer, un anticorps monoclonal humain spécifique à la protéine PAUF, et une molécule d'acide nucléique codant pour la protéine PAUF, ainsi que leurs utilisations. Étroitement corrélée à la formation du cancer, la protéine PAUF ou la molécule d'acide nucléique de PAUF peut être utilisée comme marqueur diagnostique ou cible thérapeutique. De plus, ayant la capacité de se lier spécifiquement à PAUF présentée sur la surface de diverses cellules cancéreuses, y compris les cellules cancéreuses pancréatiques, l'anticorps anti-PAUF ou son fragment fonctionnel permet le diagnostic du cancer aux stades précoces de celui-ci et l'inhibition de sa croissance, l'invasion ou la migration des cellules cancéreuses par induction de l'apoptose chez les cellules, d'où son utilité dans le diagnostic et le traitement du cancer.
PCT/KR2009/005437 2008-09-23 2009-09-23 Anticorps monoclonal humain specifique au pauf, composition pharmaceutique destinee au traitement du cancer le comprenant et procede l'utilisant pour detecter le cancer Ceased WO2010036031A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2008-0093106 2008-09-23
KR20080093106 2008-09-23

Publications (2)

Publication Number Publication Date
WO2010036031A2 true WO2010036031A2 (fr) 2010-04-01
WO2010036031A3 WO2010036031A3 (fr) 2010-06-24

Family

ID=42060277

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2009/005437 Ceased WO2010036031A2 (fr) 2008-09-23 2009-09-23 Anticorps monoclonal humain specifique au pauf, composition pharmaceutique destinee au traitement du cancer le comprenant et procede l'utilisant pour detecter le cancer

Country Status (2)

Country Link
KR (2) KR20100034720A (fr)
WO (1) WO2010036031A2 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140018304A1 (en) * 2011-03-28 2014-01-16 Industry-Academic Cooperation Foundation, Yonsei University Method for screening for a cancer treatment agent using the interaction between pauf and a binding partner thereof
CN110291110A (zh) * 2017-07-28 2019-09-27 东亚大学校产学协力团 特异性结合pauf蛋白的抗体及其用途

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012128502A2 (fr) * 2011-03-24 2012-09-27 한국생명공학연구원 Composition contenant le facteur pauf et stimulant la vasculogenèse
KR101250557B1 (ko) * 2011-05-18 2013-04-03 국립암센터 Ｐａｕｆ 특이적 앱타머 및 이를 포함하는 췌장암 치료용 조성물
WO2019022274A1 (fr) * 2017-07-28 2019-01-31 동아대학교 산학협력단 Anticorps se liant spécifiquement à une protéine pauf, et utilisation associée

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100954322B1 (ko) * 2006-06-14 2010-04-21 주식회사 엘지생명과학 췌장암과 관련된 신규한 ｌｂｆｌ313 유전자

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140018304A1 (en) * 2011-03-28 2014-01-16 Industry-Academic Cooperation Foundation, Yonsei University Method for screening for a cancer treatment agent using the interaction between pauf and a binding partner thereof
US10023915B2 (en) * 2011-03-28 2018-07-17 Industry-Academic Cooperation Foundation, Yonsei University Method for screening for a cancer treatment agent using the interaction between PAUF and a binding partner thereof
CN110291110A (zh) * 2017-07-28 2019-09-27 东亚大学校产学协力团 特异性结合pauf蛋白的抗体及其用途
AU2017425111B2 (en) * 2017-07-28 2020-06-18 Dong-A University Research Foundation For Industry-Academy Cooperation Antibody specifically binding to PAUF protein, and use thereof
CN110291110B (zh) * 2017-07-28 2023-11-24 东亚大学校产学协力团 特异性结合pauf蛋白的抗体及其用途

Also Published As

Publication number Publication date
KR20100034720A (ko) 2010-04-01
KR20110122807A (ko) 2011-11-11
KR101098186B1 (ko) 2011-12-23
WO2010036031A3 (fr) 2010-06-24

Similar Documents

Publication Publication Date Title
KR101984554B1 (ko) 췌장암의 검출 방법
WO2008051797A2 (fr) Anticorps d'agonistes anti-notch3 et leur utilisation dans le traitement de maladies liées à notch3
CN105121473A (zh) α-突触核蛋白抗体及其用途
WO2010036031A2 (fr) Anticorps monoclonal humain specifique au pauf, composition pharmaceutique destinee au traitement du cancer le comprenant et procede l'utilisant pour detecter le cancer
KR20140048303A (ko) 암의 검출 및 진단을 위한 새로운 항-cxcr4 항체 및 그의 용도
WO2017010744A1 (fr) Composition antigénique de détection d'un auto-anticorps avec réponse spécifique à la protéine exosomale eif3a, et méthode de diagnostic du cancer du foie utilisant la composition antigénique
KR20140047127A (ko) 암의 검출 및 진단을 위한 항체 i-3859의 용도
WO2022216014A1 (fr) Anticorps anti-cntn4 et son utilisation
KR20080113196A (ko) 분비 프리즐드 관련 단백질-4 (ｓｆｒｐ-4)에 대한 항체
AU2010268979B2 (en) New tumor marker
WO2019216675A1 (fr) Épitope d'antigène de surface de lymphocyte t régulateur et anticorps se liant de manière spécifique à celui-ci
WO2014021693A2 (fr) Anticorps monoclonal inédit se liant spécifiquement à la protéine tm4sf5 et son utilisation
KR101214177B1 (ko) ＣＤ66ｃ의 폐선암 특이적 에피토프 및 이를 인식하는 항체
WO2020197227A1 (fr) Biomarqueur céréblon pour le diagnostic d'un carcinome hépatocellulaire, et nouvel anticorps monoclonal spécifique de celui-ci
WO2019013392A1 (fr) Anticorps monoclonal obtenu à partir d'un antigène spécifique reconnaissant spécifiquement une protéine en2 ou composition pour le diagnostic du cancer de la prostate le contenant
WO2024196163A1 (fr) Nouvel anticorps anti-r-spondine-3 humaine et son utilisation
KR101309485B1 (ko) 비브리오 패혈증균 알티엑스에이원에 특이적인 단일클론항체, 이를 분비하는 하이브리도마 및 이를 포함하는 진단키트
WO2024196162A1 (fr) Nouveaux anticorps anti-r-spondine 2 humaine et leur utilisation
EP1856155B1 (fr) Anticorps monoclonaux et analyse d'immunodétection spécifiques pour l'épitope chimiotactique du récepteur de l'activateur du plasminogène de type urokinase
WO2022131889A1 (fr) Utilisation de protéine taci
WO2022154288A1 (fr) Anticorps monoclonal se liant de manière spécifique à la protéine kinase a d'une sous-unité catalytique alpha, et son utilisation pour le diagnostic du cancer
KR101349413B1 (ko) 비브리오 패혈증균 알티엑스에이원 항원에 결합하는 단일클론항체 제조와 진단 항원을 제공하기 위한 재조합 단백질 발현벡터
WO2010093214A2 (fr) Anticorps monoclonal reconnaissant scye1 et son utilisation
KR101309386B1 (ko) 비브리오 패혈증균 알티엑스에이원에 특이적인 단일클론항체, 이를 분비하는 하이브리도마 및 이를 포함하는 진단키트
WO2024151091A1 (fr) Composition de diagnostic et composition pharmaceutique comprenant un biomarqueur de treg dans une tumeur

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09816424

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 09816424

Country of ref document: EP

Kind code of ref document: A2