HK40017293B - Antibody capable of binding to myelin oligodendrocyte glycoprotein - Google Patents

Description

Antibodies that bind to myelin oligodendrocyte glycoproteins

Technical Field

This invention relates to antibodies or fragments of antibodies that bind to myelin oligodendrocyte glycoprotein (MOG), hybridomas that produce the antibodies or fragments, nucleic acids containing a base sequence encoding the antibody or fragment, transformed cells containing a vector containing the nucleic acid, methods for manufacturing the antibody or fragment, compositions containing the antibody or fragment, methods for detecting or measuring antigens present in the brain using the antibody or fragment, methods for diagnosing or treating brain diseases, methods for improving the brain retention of antibodies, and methods for increasing the amount of antibodies in the brain.

Background Technology

Since the initial FDA approval of muromonab-CD3 (OKT3), a mouse anti-CD3 antibody, in 1986, numerous antibody drugs have been developed. Abciximab, a chimeric antibody created by linking the variable region of a mouse antibody to the constant region of a human antibody to reduce the antigenicity of mouse antibodies, was approved in 1994.

To further reduce antigenicity, a humanized antibody technology was developed that transplants the complementarity-determining region (CDR) of the variable region of a mouse antibody, which plays an important role in antigen binding, into the framework region (FR) of a human antibody. The humanized anti-CD20 antibody dacizumab was approved in 1997.

In addition, phage display technology using human antibody sequence libraries has been applied. As the first antibody obtained using phage display technology, the fully human anti-TNFα antibody adalimumab was approved in 2002. More than 60 antibody drugs targeting CD20, CD52, TNFα, HER2, EGFR, etc., have been approved (Non-Patent Literature 1).

Thus, antibodies have become a widely known form of medicine. To date, the majority of approved antibody drugs target cancer and immune diseases, accounting for over 75% of the total.

In the treatment of central nervous system diseases, the importance of biological agents such as antibodies is also increasing. In animal models, studies have been reported on the use of monoclonal antibodies against β-amyloid protein in Alzheimer's disease; and various neurotrophic factors with neuroprotective effects (brain-derived neurotrophic factor, BDNF; glial-derived neurotrophic factor, GDNF) have shown neuroprotective effects in central nervous system diseases (Non-Patent Literature 2).

However, when antibodies are administered peripherally, the delivery rate to the central nervous system is lower than that to other organs, with reported antibody migration rates (the ratio of concentration in cerebrospinal fluid (CSF) to serum concentration) of 0.1-0.3% (Non-Patent Literature 3-5).

One reason for the reduced drug delivery in the central nervous system, including the brain and spinal cord, is the mechanism by which the blood-brain barrier (BBB) restricts the transport of substances between the blood and the tissue fluid of the brain. The BBB possesses both a physical, non-specific control mechanism based on intercellular connections in vascular endothelial cells and a substrate-specific efflux mechanism based on efflux transporters. It protects the central nervous system from the effects of foreign substances or drugs and plays a crucial role in maintaining homeostasis.

However, due to the presence of the blood-brain barrier, it is difficult to achieve effective drug concentrations in the central nervous system, making drug development challenging. For example, enzyme supplementation therapy has been implemented through intravenous administration of α-L-iduronase for Henle syndrome (mucopolysaccharidosis type I) and iduronate 2-sulfatase for Hunter syndrome (mucopolysaccharidosis type II), but due to the large molecular weight of the enzymes, they cannot cross the blood-brain barrier, and therefore their effectiveness against central nervous system symptoms has not been confirmed (Non-Patent Literature 6-9). Furthermore, side effects such as the production of neutralizing antibodies have been reported due to periodic, continuous administration of a certain amount of recombinant enzymes (Non-Patent Literature 10).

In addition, attempts have been made to directly administer biological agents into the medullary cavity or brain to increase intracerebral concentration. For example, a method for administering iduronate 2-sulfatase into the brain of patients with Hunter syndrome (mucopolysaccharidosis type II) has been reported (Patent Document 1). However, direct administration into the medullary cavity or brain is highly invasive (Non-Patent Document 11).

Therefore, various delivery technologies have been studied to increase the intracerebral concentration of macromolecules such as biological agents. For example, several methods have been reported to bind to membrane proteins expressed on cerebral vascular endothelial cells to form complexes of macromolecules and membrane proteins, and to facilitate their crossing of the blood-brain barrier via endocytosis.

Most of the reported techniques utilize receptor-mediated transcytosis (RMT) to target brain vascular endothelial receptors, such as transferrin receptor, insulin receptor, insulin-like growth factor receptor, and low-density lipoprotein receptor family (LDLRf).

A transferrin receptor-mediated blood-brain barrier crossing technique was reported by creating a fusion protein of an anti-transferrin receptor antibody and nerve growth factor. As techniques using anti-transferrin receptor antibodies, bispecific antibodies of anti-transferrin receptor antibody and anti-β-secretase (BACE1) antibody were reported (Patent Documents 2 and 3 and Non-Patent Documents 12 and 13), and fusion antibodies formed by fusing an anti-transferrin receptor monovalent antibody to the carboxyl terminus of an anti-amyloid β antibody (Patent Document 4, Non-Patent Document 14).

Regarding the brain delivery of bispecific antibodies against transferrin receptor and BACE1, it has been reported that when the antibodies were administered at a dose of 20 mg/kg body weight in mice, the amount of antibody uptake in the brain increased to approximately four times that of the control (Non-Patent Literature 13).

In addition, a technique for enabling drugs to cross the blood-brain barrier by encapsulating the drug within liposomes with anti-transferrin receptor antibodies on their surface has been reported. It has been reported that using a fusion of anti-rat transferrin receptor antibodies and immune micelles, the uptake in the rat brain increased by approximately 2 to 5 times (Non-Patent Literature 9).

In addition, insulin receptor-mediated blood-brain barrier crossing technology has been reported by creating fusion proteins by fusing neurotrophic factors, enzymes, or anti-amyloid antibodies to the carboxyl terminus of anti-insulin receptor antibodies (Non-Patent Literature 16-19).

According to reports, in rhesus monkeys, the uptake in the brain of a fusion antibody labeled with anti-human insulin receptor and GDNF was about 15 times that of GDNF 2 hours after administration (Non-Patent Literature 17).

However, transferrin receptors and insulin receptors are expressed not only in brain vascular endothelial cells but also throughout the body, including in the liver. Therefore, these technologies can increase drug delivery to the central nervous system while also delivering drugs to the liver and other organs (Non-Patent Literature 20). Furthermore, because the antigen is expressed throughout the body, the antibody has a short blood half-life (Non-Patent Literature 12).

In addition, an antibody (Fc5) targeting the brain vascular endothelial membrane expression antigen TMEM30A has been reported to exhibit RMT-like activity (Patent Document 5 and Non-Patent Documents 21 and 22). Fc5 is a variable domain of heavy chain antibody (hereinafter referred to as VHH) antibody derived from a single domain of a llama heavy chain antibody. In in vitro BBB model and in vivo rat model, increased brain delivery of the fusion of Fc5 and human Fc was shown compared with control IgG.

According to reports, the fusion of Fc5-derived single-chain antibody (scFv) and metabotropic glutamate receptor type I (mGluRI) antibody increased CSF exposure in rat models by approximately 5-fold compared to the control fusion of single-chain antibody and mGluRI antibody (Non-Patent Literature 23).

In addition, it has been reported that IgG antibodies are rapidly expelled from the brain into the circulating blood via neonatal Fc receptors (Non-Patent Literature 24 and 25), for example, the brain half-life of IgG in rats after intracerebral administration is as short as 48 minutes (Non-Patent Literature 24).

MOGs are proteins belonging to the immunoglobulin superfamily that constitute myelin. Human MOGs consist of 218 amino acids in total and are expressed in the outermost layer of myelin in the central nervous system, playing a role in cell adhesion and cell surface interactions (Non-Patent Literature 26-28).

In inflammatory diseases such as multiple sclerosis (MS), where glial cells in the central nervous system are attacked by autoimmunity, MOG is considered a candidate autoantigen (Non-Patent Literature 29 and 30). Low concentrations of anti-MOG antibodies in serum have been reported in MS patients, but they have also been detected in the central nervous system (Non-Patent Literature 29).

As a result, the following reports exist: In pathological conditions such as MS, the blood-brain barrier is disrupted due to leakage of humoral factors and invasion of inflammatory cells, making it easier for antibodies to migrate into the central nervous system (Non-Patent Literature 30 and 31). Furthermore, reports have also indicated the local generation of autoantibodies within the central nervous system through infiltration of B cells and plasma cells (Non-Patent Literature 30, 32, and 33).

Experimental autoimmune encephalomyelitis (EAE) shares many pathological features with MS, and is therefore a model used in MS pathology studies. It has been reported that EAE can be induced in animals by immunizing them with MOG proteins or peptides (Non-Patent Literature 34).

Furthermore, there are reports of EAE scores worsening after administration of anti-MOG antibodies to animals induced with EAE (Non-Patent Literature 29 and 35), but peak EAE scores were observed 1-2 days after antibody administration (Non-Patent Literature 29) or 4 days after administration (Non-Patent Literature 35), indicating that the peaks were transient. On the other hand, it has been reported that EAE does not occur in normal animals even after administration of anti-MOG antibodies alone (Non-Patent Literature 36 and 37).

Existing technical documents

Patent documents

Patent Document 1: International Publication No. 2012/023623

Patent Document 2: International Publication No. 2016/081640

Patent Document 3: International Publication No. 2016/081643

Patent Document 4: International Publication No. 2014/033074

Patent Document 5: Specification of Canadian Patent No. 2623841

Non-patent literature

Non-patent literature 1: Kyla RR. and Richard CC., Biotechnol Adv., pii:S0734-9750(16),30091-X,2016

Non-patent literature 2: Pardridge WM., Bioconjugate Chem., 19, 1327-1338, 2008

Non-patent literature 3: Wang W., et al., Clin. pharmacol. Ther., 84, 548-558, 2008

Non-patent literature 4: Garg A., et al., AAPSJ., 11, 553-557, 2009

Non-patent literature 5: Kaj B., et al., Arch. Neurol., 69(8), 1002-1010, 2012

Non-patent literature 6: Wraith JE. et al., J. Pediatr. 144(5), 581-588, 2004

Non-patent literature 7: Muenzer J. et al., Genet Med. 8(8), 465-473, 2006

Non-Patent Document 8: ALDURAZYME (Registered Trademark) Intravenous Drip Solution 2.9mg with Accompanying Information (8th Edition, July 2016)

Non-Patent Document 9: ELAPRASE (Registered Trademark) Intravenous Drip Solution 6mg with Accompanying Information (6th Edition, July 2016)

Non-patent literature 10: Brooks, D.A. et al., Trends Mol. Med. 9, 450-453, 2003

Non-patent literature 11: Sorrentino NC. et al., Pediatr Endocrinol Rev. 1, 630-638, 2016

Non-patent literature 12: Couch JA., et al., Science Translational Medicine, 5, 183ra57, 2013

Non-patent literature 13: Yu YJ., et al., Science Translational Medicine, 6, 261ra154, 2014

Non-patent literature 14: Niewoehner J., et al., Neuron. 81, 49-60, 2014

Non-patent literature 15: Jun Y., et al., Macromol. Biosci. 12, 1209-1219, 2012

Non-patent literature 16: Pardridge WM. and Boado RJ., Methods in Enzymology, 503, 269-292, 2012

Non-patent literature 17: Boado RJ., et al., Drug Metab. Dispos., 37(12), 2299-2304, 2009

Non-patent literature 18: Boado RJ., et al., J. Pharmacol. Exp. Ther., 333(3), 961-969, 2010

Non-patent literature 19: Boado RJ., et al., Bioconjugate Chem., 1, 97-104, 2012

Non-patent literature 20: Yun Zhang et al., J. Pharmacol. Exp. Ther., 313(3), 1075-1081, 2005

Non-patent literature 21: Abulrob A., et al., J. Neuyrochem., 95(4), 1201-1214, 2005

Non-patent literature 22: Farrington GK., et al., FASEB J., 28, 4764-4778, 2014

Non-patent literature 23: Webster CI., et al., FASEB J., 30, 1927-1940, 2016

Non-patent literature 24: Zhang Y., et al., J. Neuroimmunol., 114(1-2), 168-172, 2001

Non-patent literature 25: Philip RC., et al., Brain Research, 1534, 13-21, 2013

Non-patent literature 26: Brunner C., et al., J. Neurochem, 52, 296-394, 1989

Non-Patent Literature 27: Pham-Dinh D., et al., Proc. Natl. Acad. Sci. USA, 90, 7990-7994, 1993

Non-patent literature 28: Gardinier MV., et al., J. Neurosci. Res., 33, 177-187, 1992

Non-patent literature 29: Eduard Urich, et al., PNAS, 103, 18697-18702, 2006

Non-patent literature 30: Markus Reindl, et al., Brain, 122, 2047-2056, 1999

Non-patent literature 31: Shimizu F., et al., Nihon Rinsho. 72(11), 1949-1954, 2014

Non-patent literature 32: Nese Sinmaz., et al., Ann. N. Y. Acad. Sci., 1351, 22-38, 2015

Non-patent literature 33: F.J. Quintana, Neurology, 78, 532-539, 2012

Non-patent literature 34: Ralf Gold., et al., Brain, 129, 1953-1971, 2006

Non-patent literature 35: Margaret M., et al., J. Neuroimmunology, 125, 114-124, 2002

Non-patent literature 36: G. Locatelli, et al., Nature Neuro Science, 15(4), 543-551, 2012

Non-patent literature 37: H J Schluesener, et al., J. Immunol., 139, 4016-4021, 1987

Summary of the Invention

The problem that the invention aims to solve

Non-patent documents 29 and 35 describe the detection of anti-MOG antibodies in the brain when administered to EAE models, but there are no reports of detecting anti-MOG antibodies in the brain when administered peripherally to normal animals.

This invention relates to MOG-binding molecules that bind to myelin oligodendrocyte glycoprotein (MOG) and methods of using such molecules. Specifically, the object of this invention is to provide an antibody or antibody fragment that binds to MOG, a hybridoma that produces the antibody or antibody fragment, a nucleic acid containing a base sequence encoding the antibody or antibody fragment, transformed cells containing a vector comprising the nucleic acid, a method for manufacturing the antibody or antibody fragment, a composition containing the antibody or antibody fragment, and methods for detecting or measuring antigens present in the brain using the antibody or antibody fragment, methods for diagnosing or treating brain diseases, methods for improving the brain retention of antibodies, and methods for increasing the amount of antibodies in the brain.

Methods for solving problems

As a method for solving the above problems, the present invention provides a MOG-binding molecule that binds to MOG and a method for using the molecule, specifically, providing an antibody or a fragment of the antibody.

That is, the present invention relates to items (1) to (22) below.

(1) An antibody or a fragment thereof that binds to a myelin oligodendrocyte glycoprotein (hereinafter referred to as MOG).

(2) The antibody or the antibody fragment as described in (1), wherein the antibody has brain retention properties.

(3) The antibody or antibody fragment as described in (1) or (2), wherein the antibody is selected from the group consisting of (a) to (r) below.

(a) An antibody in which the amino acid sequences of the complementarity-determining regions (CDRs) 1 to 3 of the heavy chain variable region (hereinafter referred to as VH) contain the amino acid sequences described in sequence numbers 4, 5 and 6, respectively, and the amino acid sequences of the CDRs 1 to 3 of the light chain variable region (VL) contain the amino acid sequences described in sequence numbers 10, 11 and 12, respectively.

(b) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 16, 17 and 18, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 22, 23 and 24, respectively.

(c) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 28, 29 and 30, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 34, 35 and 36, respectively.

(d) The amino acid sequences of CDR1 to 3 of the heavy chain variable region (hereinafter referred to as VHH) of the heavy chain antibody contain the antibody fragments containing the amino acid sequences recorded in sequence numbers 40, 41 and 42, respectively.

(e) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 153, 154 and 155, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 158, 159 and 160, respectively.

(f) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 163, 164 and 165, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 168, 169 and 170, respectively.

(g)Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 173, 174 and 175, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 178, 179 and 180, respectively.

(h)Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 183, 184 and 185, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 188, 189 and 190, respectively.

(i) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 193, 194 and 195, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 198, 199 and 200, respectively.

(j) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 203, 204 and 205, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 208, 209 and 210, respectively.

(k)Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 213, 214 and 215, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 218, 219 and 220, respectively.

(l) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 223, 224 and 225, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 228, 229 and 230, respectively.

(m)Antibodies whose CDR1-3 amino acid sequences of VH contain the amino acid sequences described in sequence numbers 233, 234 and 235, respectively, and whose CDR1-3 amino acid sequences of VL contain the amino acid sequences described in sequence numbers 238, 239 and 240, respectively.

(n) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 243, 244 and 245, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 248, 249 and 250, respectively.

(o) An antibody that competitively binds to MOG with at least one of the antibodies described in (a) to (n) above;

(p) An antibody that binds to an epitope containing any of the antibodies described in (a) to (n) above;

(q) An antibody that binds to the same epitope as any of the antibodies described in (a) to (n) above; and

(r) An antibody that contains an amino acid sequence having more than 85% homology with the amino acid sequence of any of the antibodies described in (a) to (n) above.

(4) The antibody or antibody fragment as described in any one of (1) to (3), wherein the antibody is selected from the group consisting of (a) to (n), (o1) to (o22) and (p).

(a) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 3 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 9.

(b) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 15 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 21.

(c) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 27 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 33.

(d) The amino acid sequence of VHH contains an antibody fragment containing the amino acid sequence described in sequence number 39.

(e)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 152 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 157.

(f) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 162 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 167.

(g)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 172 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 177.

(h)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 182 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 187.

(i) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 192 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 197.

(j) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 202 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 207.

(k)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 212 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 217.

(l)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 222 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 227.

(m)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 232 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 237.

(n)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 242 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 247.

(o1)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 252 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 254.

(o2)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 256 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 258.

(o3)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 260 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 262.

(o4)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 264 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 266.

(o5)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 268 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 270.

(o6)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 272 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 274.

(o7)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 276 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 278.

(o8)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 280 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 282.

(o9)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 284 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 286.

(o10)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 288 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 290.

(o11)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 292 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 294.

(o12)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 296 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 298.

(o13)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 300 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 302.

(o14)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 304 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 306.

(o15)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 308 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 310.

(o16)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 312 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 314.

(o17)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 316 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 318.

(o18)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 320 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 322.

(o19)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 324 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 326.

(o20)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 328 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 330.

(o21)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 332 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 334.

(o22)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 336 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 338.

(p) An antibody that contains an amino acid sequence having more than 85% homology to the amino acid sequence of any of the antibodies described in (a) to (n) and (o1) to (o22) above.

(5) The antibody or antibody fragment as described in any one of (1) to (4), wherein the antibody or antibody fragment is a bispecific antibody.

(6) The bispecific antibody as described in (5), wherein the bispecific antibody binds to MOG and antigens present in the brain.

(7) A bispecific antibody as described in (5) or (6), wherein the bispecific antibody comprises an antigen-binding site that binds to the MOG and an antigen-binding site that binds to an antigen present in the brain.

(8) The antibody fragment as described in any one of (1) to (7), wherein the antibody fragment is selected from the group consisting of Fab, Fab', F(ab') ₂ , single-chain antibody (scFv), dimerized V region (double-chain antibody (diabody)), disulfide bond-stabilized V region (dsFv), VHH and peptides containing CDR.

(9) The antibody and the antibody fragment as described in any one of (1) to (8), wherein the antibody is a recombinant antibody.

(10) The antibody and the antibody fragment as described in any one of (1) to (9), wherein the antibody is selected from the group consisting of mouse antibody, rat antibody, rabbit antibody, alpaca antibody, camel antibody, llama antibody, chimeric antibody, humanized antibody and human antibody.

(11) A fusion antibody or fusion antibody fragment formed by binding at least one of the group consisting of (a) to (c) below with any one of (1) to (10) an antibody or the antibody fragment that binds to MOG.

(a) Hydrophilic polymers,

(b) Amphiphilic macromolecules, and

(c) Functional molecules.

(12) A hybridoma that produces any one of (1) to (11) antibodies.

(13) A nucleic acid comprising a base sequence encoding an antibody as described in any one of (1) to (11).

(14) A transformed cell containing a carrier comprising the nucleic acid described in (13).

A method for manufacturing the antibody or antibody fragment described in any one of (15) (1) to (11), comprising: culturing the hybridoma described in (12) or the transformed cell described in (14), and collecting the antibody or antibody fragment described in any one of (1) to (11) from the culture medium.

(16) A composition comprising the antibody or the antibody fragment of any one of (1) to (11).

(17) The composition as described in (16) is a composition for detecting or determining antigens present in the brain.

(18) The composition as described in (16) is a composition for the diagnosis or treatment of brain diseases.

(19) A method for detecting or determining an antigen present in the brain, wherein the antigen present in the brain is detected or determined using any one of (1) to (11) or an antibody fragment thereof, or the composition described in (16).

(20) A method for diagnosing or treating a brain disease, wherein the brain disease is diagnosed or treated using any one of (1) to (11) or an antibody fragment thereof, or the composition described in (16).

(21) A method for improving the brain retention of an antibody or antibody fragment or fusion antibody or fusion antibody fragment, wherein the antibody or antibody fragment or fusion antibody or fusion antibody fragment described in any one of (1) to (11), or the composition described in (16) is used to improve the brain retention of the antibody or antibody fragment or fusion antibody or fusion antibody fragment.

(22) A method for increasing the amount of antibodies or antibody fragments or fusion antibodies or fusion antibody fragments in the brain, wherein the amount of antibodies or antibody fragments or fusion antibodies or fusion antibody fragments in the brain is increased by using any one of (1) to (11), or the composition of (16).

Invention Effects

The MOG-binding molecule of the present invention not only increases the brain retention of the molecule itself by specifically binding to MOGs, but also enables its transport and retention in the brain by modifying other molecules onto the MOG-binding molecule, thereby enabling its application in the treatment of brain diseases. Antibodies are a specific example of the MOG-binding molecule of the present invention. The antibody or antibody fragment of the present invention is an antibody that exhibits brain retention by binding to MOGs in the brain. Therefore, the antibody or antibody fragment of the present invention can be used as a composition for detecting or measuring antigens (MOGs, or MOGs and other antigens present in the brain) present in the brain, a composition for diagnosing brain diseases, and a pharmaceutical composition for treating brain diseases.

Attached Figure Description

Figure 1 shows the results of ELISA analysis of scFv antibodies bound to MOG, demonstrating the binding affinity of phage clones to rMOG-FLAG_Fc. The vertical axis represents the absorbance of rMOG-FLAG_Fc, and the horizontal axis represents the name of the scFv antibody displayed by each phage clone.

Figure 2 shows the results of flow cytometry analysis of the binding of various anti-MOG antibodies to HEK cells, rat MOG/HEK cells, mouse MOG/HEK cells, cynomolgus monkey MOG/HEK cells, or human MOG/HEK cells. The vertical axis represents the cell number, and the horizontal axis represents the fluorescence intensity. The dashed histogram shows the binding of the anti-AVM antibody used as a negative control, and the solid histogram shows the binding of each MOG antibody.

Figures 3(A) and (B) show the results of the rat brain migration evaluation of anti-MOG antibodies. Figure 3(A) shows the antibody concentration in serum 4 days after administration of the antibody to rats. The vertical axis represents the antibody concentration (ng/mL), and the horizontal axis represents the administered antibody. Figure 3(B) shows the antibody concentration in brain tissue 4 days after administration of the antibody to rats. The vertical axis represents the amount of antibody per unit brain weight (ng/g brain), and the horizontal axis represents the administered antibody. In either figure, white bars represent anti-AVM antibodies used as negative controls, and black bars represent anti-MOG antibodies.

Figures 4(A) and (B) show the results of the rat brain migration evaluation of anti-MOG antibodies. Figure 4(A) shows the antibody concentration in serum 4 and 10 days after antibody administration to rats. The vertical axis represents antibody concentration (ng/mL), and the horizontal axis represents the number of days after antibody administration. Figure 4(B) shows the antibody concentration in brain tissue 4 and 10 days after antibody administration to rats. The vertical axis represents the amount of antibody per unit brain weight (ng/g brain), and the horizontal axis represents the number of days after antibody administration. In either figure, white diamonds represent anti-AVM antibodies used as negative controls, white quadrilaterals represent anti-transferrin receptor antibody OX26, and black triangles represent anti-MOG antibody MOG01.

Figure 5 shows the results of flow cytometry analysis of the binding of various bispecific antibodies to HEK293F cells, rat MOG/HEK293F cells, or human MOG/HEK293F cells. The vertical axis represents the cell number, and the horizontal axis represents the fluorescence intensity. The dashed histogram shows the binding of the anti-AVM antibody used as a negative control, and the solid histogram shows the binding of each bispecific antibody.

Figure 6 shows the results of flow cytometry analysis of the binding of various bispecific antibodies to the human breast cancer cell line SK-BR-3. The vertical axis represents the cell number, and the horizontal axis represents the fluorescence intensity. The dashed histogram shows the binding of the anti-AVM antibody used as a negative control, and the solid histogram shows the binding of each bispecific antibody.

Figures 7(A) and (B) show the results of evaluating the migration of the bispecific antibody bound to MOG in rat brains. Figure 7(A) shows the antibody concentration in serum 10 days after administration of the antibody to rats. The vertical axis represents the antibody concentration (ng/mL), and the horizontal axis represents the bispecific antibody used. Figure 7(B) shows the antibody concentration in brain tissue 10 days after administration of the antibody to rats. The vertical axis represents the amount of antibody per unit brain weight (ng/g brain), and the horizontal axis represents the bispecific antibody used.

Figures 8(A) and (B) show the results of evaluating the migration of anti-MOG01 antibody in mouse brains. Figure 8(A) shows the antibody concentration in serum of mice 3, 6, 10, 14, 21, and 28 days after administration of the antibody. The vertical axis represents antibody concentration (ng/mL), and the horizontal axis represents time (days). Figure 8(B) shows the antibody concentration in brain tissue of mice 3, 6, 10, 14, 21, and 28 days after administration of the antibody. The vertical axis represents antibody concentration (ng/g brain), and the horizontal axis represents time (days). In either figure, white circles represent anti-AVM antibody used as a negative control, and black quadrilaterals represent MOG01scFv-hG4PE.

Figures 9(A)–(C) show the results of brain migration imaging evaluation of the anti-MOG01 antibody in mice. Figure 9(A) shows the fluorescence intensity data of the brain 6 days after administration of Alexa FluorR 488-labeled anti-AVM antibody and Alexa FluorR 488-labeled anti-MOG01 antibody as negative controls. Figure 9(B) shows the fluorescence intensity data of the brain 14 days after administration. Figure 9(C) shows the values of fluorescence intensity in the brain after 6 and 14 days after administration, corrected for the fluorescence intensity of the administered antibody. The vertical axis represents the fluorescence intensity in the brain/fluorescence intensity of the administered antibody (%), and the horizontal axis represents the administered antibody.

Figures 10(A)–(C) show the structures of various bispecific antibodies that bind to AVM and MOG. Figure 10(A) shows the structure of the AVM-MOG01 IgG4PE(R409K) antibody, Figure 10(B) shows the structure of the AVM IgG4PE(R409K)_MOG01Fab antibody, and Figure 10(C) shows the structure of the AVM IgG4PE(R409K)_MOG01sscFv antibody.

Figures 11(A) and (B) show the structures of various bispecific antibodies that bind to AVM and MOG. Figure 11(A) shows the structures of AVMIgG4PE(R409K)_MOG01dscFv antibody, AVM IgG4PE(R409K)_MOG01dscFv2 antibody, and AVM IgG4PE(R409K)_MOG01dscFv4 antibody, and Figure 11(B) shows the structures of AVM IgG4PE(R409K)_MOG01dscFv3 antibody, AVM IgG4PE(R409K)_MOG01dscFv5 antibody to AVM IgG4PE(R409K)_MOG01dscFv11 antibody.

Figures 12(A)–(C) show the results of flow cytometry analysis of the binding of various bispecific antibodies to human MOG/L929 cells. The vertical axis represents the average fluorescence intensity, and the horizontal axis represents the antibody concentration. In Figure 12(A), white circles represent AVMIgG4PE(R409K) antibody (negative control), and black quadrilaterals represent AVM-MOG01 IgG4PE(R409K) antibody. In Figure 12(B), white circles represent AVMIgG4PE(R409K)_AVMsscFv antibody (negative control), and black quadrilaterals represent AVMIgG4PE(R409K)_MOG01sscFv antibody. In Figure 12(C), white circles represent AVMIgG4PE(R409K)_AVMFab antibody (negative control), and black quadrilaterals represent AVMIgG4PE(R409K)_MOG01Fab antibody.

Figures 13(A) and (B) show the results of flow cytometry analysis of the binding of various bispecific antibodies to human MOG/L929 cells. The vertical axis represents the average fluorescence intensity, and the horizontal axis represents the antibody concentration. In Figure 13(A), the white quadrilaterals represent AVMIgG4PE(R409K)_MOG01dscFv antibody, the white circles represent AVMIgG4PE(R409K)_MOG01dscFv2 antibody, and the white triangles represent AVMIgG4PE(R409K)_MOG01dscFv4 antibody. In Figure 13(B), the white rhombus represents AVMIgG4PE(R409K)_MOG01dscFv3 antibody, the black rhombus represents AVMIgG4PE(R409K)_MOG01dscFv5 antibody, the white circle represents AVMIgG4PE(R409K)_MOG01dscFv6 antibody, the black circle represents AVMIgG4PE(R409K)_MOG01dscFv7 antibody, the white triangle represents AVMIgG4PE(R409K)_MOG01dscFv8 antibody, the black triangle represents AVMIgG4PE(R409K)_MOG01dscFv9 antibody, the white quadrilateral represents AVMIgG4PE(R409K)_MOG01dscFv10 antibody, and the black quadrilateral represents AVMIgG4PE(R409K)_MOG01dscFv11 antibody.

Figures 14(A) and (B) show the results of mouse brain migration evaluation for various bispecific antibodies. The vertical axis represents antibody concentration, and the horizontal axis represents the bispecific antibody used. Figures 14(A) and (B) show the serum and brain tissue antibody concentrations 10 days after administration of AVM IgG4PE(R409K) antibody (negative control) and AVM-MOG01IgG4PE(R409K) antibody, respectively.

Figures 15(A) and (B) show the results of mouse brain migration evaluation for various bispecific antibodies. The vertical axis represents antibody concentration, and the horizontal axis represents the bispecific antibody used. Figures 15(A) and (B) show the serum and brain tissue antibody concentrations 10 days after administration of AVM IgG4PE(R409K)_AVMsscFv antibody (negative control) and AVM IgG4PE(R409K)_MOG01sscFv antibody, respectively.

Figures 16(A) and (B) show the results of mouse brain migration evaluation for various bispecific antibodies. The vertical axis represents antibody concentration, and the horizontal axis represents the bispecific antibody used. Figures 16(A) and (B) show the serum and brain tissue antibody concentrations 10 days after administration of AVM IgG4PE(R409K)_AVMFab antibody (negative control) and AVM IgG4PE(R409K)_MOG01Fab antibody, respectively.

Figures 17(A)–(D) show the results of mouse brain migration evaluation for various bispecific antibodies. The vertical axis represents antibody concentration, and the horizontal axis represents the bispecific antibody used. The negative control corresponding to the AVM IgG4PE(R409K)_MOG01dscFv antibody is the AVM IgG4PE(R409K)_AVMdscFv antibody; the negative control corresponding to the AVM IgG4PE(R409K)_MOG01dscFv3 antibody is the AVM IgG4PE(R409K)_AVMdscFv3 antibody; and the negative control corresponding to the AVM IgG4PE(R409K)_MOG01dscFv5 antibody is the AVM IgG4PE(R409K)_AVMdscFv5 antibody. Figure 17(A) shows the antibody concentration in serum 10 days after antibody administration. Figure 17(B) shows the antibody concentration in brain tissue 10 days after antibody administration. Figure 17(C) shows the antibody concentration in serum 28 days after antibody administration. Figure 17(D) shows the antibody concentration in brain tissue 28 days after antibody administration.

Figure 18 shows the amino acid sequence of scFv of a similar clone of the MOG antibody, illustrating a similar clone of the MOG301 antibody.

Figure 19 shows the amino acid sequence of scFv of a similar clone of the MOG antibody, illustrating a similar clone of the MOG303 antibody.

Figure 20 shows the amino acid sequence of scFv of a similar clone of the MOG antibody, illustrating a similar clone of the MOG307 antibody.

Figure 21 shows the amino acid sequence of scFv of a similar clone of the MOG antibody, illustrating a similar clone of the MOG310 antibody.

Figures 22(A) and (B) show the amino acid sequences of scFv of similar clones of the MOG antibody. Figure 22(A) shows a similar clone of the MOG329 antibody, and Figure 22(B) shows a similar clone of the MOG456 antibody.

Figure 23 shows the results of flow cytometry analysis of the binding of anti-MOG antibodies to Expi293F cells. The vertical axis represents cell number, and the horizontal axis represents fluorescence intensity. The dashed histogram shows the binding of anti-AVM antibody used as a negative control, and the solid histogram shows the binding of each MOG antibody.

Figure 24 shows the results of flow cytometry analysis of the binding of anti-MOG antibodies to mouse MOG/Expi293F cells. The vertical axis represents cell number, and the horizontal axis represents fluorescence intensity. The dashed histogram shows the binding of anti-AVM antibody used as a negative control, and the solid histogram shows the binding of each MOG antibody.

Figure 25 shows the results of flow cytometry analysis of the binding of anti-MOG antibodies to human MOG/Expi293F cells. The vertical axis represents cell number, and the horizontal axis represents fluorescence intensity. The dashed histogram shows the binding of anti-AVM antibody used as a negative control, and the solid histogram shows the binding of each MOG antibody.

Figure 26 shows the results of flow cytometry analysis of the binding of the enzyme fusion antibody MOG01IgG4PE(R409K)-ASM to human MOG/L929 cells. The vertical axis represents the average fluorescence intensity, and the horizontal axis represents the antibody concentration.

Figure 27 shows the results of ELISA analysis of the binding of anti-ASM antibodies (manufactured by LSBio) to MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM. The vertical axis represents absorbance, and the horizontal axis represents the name of the antibody on the immobilized phase. MOG01 IgG4PE and AVM IgG4PE were used as negative controls. Thin diagonal bars indicate data for anti-ASM antibody at 5 μg/mL, thick diagonal bars indicate data for anti-ASM antibody at 1 μg/mL, and white bars indicate data for anti-ASM antibody at 0.2 μg/mL.

Figures 28(A) and (B) show the results of mouse brain migration evaluation of the enzyme fusion antibodies MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM. The vertical axis represents antibody concentration, and the horizontal axis represents the enzyme fusion antibody used. Figure 28(A) shows the antibody concentration in serum 10 days after antibody administration. Figure 28(B) shows the antibody concentration in brain tissue 10 days after antibody administration.

Detailed Implementation

This invention relates to antigen-binding molecules that bind to myelin-oligodendrocyte glycoprotein (hereinafter referred to as MOG). More specifically, this invention relates to antibodies or fragments of antibodies that bind to MOG.

As the MOG-binding molecule of the present invention, any molecule that specifically binds to MOG and remains in the brain can be of any molecular form, including proteins, nucleic acids, and low- or high-molecular-weight organic compounds. Specifically, it can be any molecule such as recombinant proteins, antibodies, aptamers, or low-molecular-weight compounds obtained through low-molecular-weight screening; antibodies and antibody fragments are preferred examples. The MOG-binding molecule is preferably a molecule that binds to the extracellular region of MOG.

MOGs are proteins belonging to the immunoglobulin superfamily that constitute myelin. For example, the human MOG consists of 218 amino acids and is expressed in the outermost layer of the myelin sheath in the central nervous system, playing a role in cell adhesion and cell surface interactions.

The animal species to which the MOG binding molecule of the present invention binds can include mice, rats, cynomolgus monkeys, and/or humans, but are not particularly limited to these species. An appropriate animal species can be selected based on the intended use of the antibody. For example, when using the antibody of the present invention for human pharmaceutical purposes, the antibody is preferably an antibody that binds to at least a human MOG.

In this invention, examples of human MOGs include: polypeptides comprising the amino acid sequence described in Serial No. 78 or the amino acid sequence of NCBI Registry No. AAB08088; polypeptides consisting of an amino acid sequence obtained by deleting, substituting, or adding one or more amino acids in the amino acid sequence described in Serial No. 78 or the amino acid sequence of NCBI Registry No. AAB08088 and having the function of a human MOG; or polypeptides consisting of an amino acid sequence having 60% or more, preferably 80% or more, further preferably 90% or more, and most preferably 95% or more homology with the amino acid sequence described in Serial No. 78 or the amino acid sequence of NCBI Registry No. AAB08088 and having the function of a human MOG.

Peptides with an amino acid sequence obtained by deleting, substituting, or adding one or more amino acids in the amino acid sequence described in sequence number 78 or represented by NCBI accession number AAB08088 can be processed using site-directed mutagenesis [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989); Current Protocols in Molecular Biology, John Willie & Son Press (1987-1997); Nucleic Acids Research, 10, 6487 (1982); Proc. Natl. Acad. Sci. USA, 79, 6409 (1982); Gene, 34, 315 (1985); Nucleic Acids Research, 13, 4431 (1985); Proc. Natl. Acad. Sci. USA, 82, 488 (1985) etc., obtained by introducing site-directed mutations into DNA encoding, for example, a polypeptide containing the amino acid sequence of sequence number 78.

The number of amino acids that are missing, substituted, or added is not particularly limited, but is preferably 1 to several tens, for example 1 to 20, and more preferably 1 to several, for example 1 to 5 amino acids.

The same applies to the amino acid sequences of mouse MOG [Sequence No. 74, NCBI Accession No. NP_034944], rat MOG [Sequence No. 68, NCBI Accession No. AAA41628], and cynomolgus monkey MOG [Sequence No. 76, NCBI Accession No. NP_001271785].

Genes encoding human MOGs include the base sequences described in Serial No. 77 and the NCBI accession number U64564. In this invention, genes encoding MOGs include: genes comprising DNA sequences obtained by deleting, substituting, or adding one or more bases from the base sequences described in Serial No. 77 or the NCBI accession number U64564, encoding a polypeptide with MOG function; genes comprising DNA sequences having at least 60% homology with the base sequences described in Serial No. 77 or the NCBI accession number U64564, preferably having at least 80% homology, and more preferably having at least 95% homology, encoding a polypeptide with MOG function; and genes comprising DNA that hybridizes under stringent conditions with DNA containing the base sequences described in Serial No. 77 or the NCBI accession number U64564, encoding a polypeptide with MOG function.

DNA that hybridizes under strict conditions refers to DNA capable of hybridization obtained by using DNA containing the base sequence described in sequence number 77 or the base sequence of NCBI accession number U64564 as a probe, such as colony hybridization, plaque hybridization, Southern blotting, or DNA microarray.

Specifically, the following methods can be used to identify DNA: using a filter membrane or slide immobilized with DNA derived from hybridization colonies or plaques, or PCR products or oligoDNA containing the sequence, hybridization is performed at 65°C in the presence of 0.7–1.0 mol/L sodium chloride [Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press (1989); Current Protocols in Molecular Biology, John Willie & Son (1987–1997); DNA Cloning 1: Core Techniques, A Practical Approach, 2nd Edition, Oxford University Press (1995)]. The filter membrane or slide is then washed at 65°C with 0.1–2 times the concentration of SSC solution (1 times the concentration of SSC solution consists of 150 mmol/L sodium chloride and 15 mmol/L sodium citrate).

As for DNA capable of hybridization, examples include DNA that has at least 60% homology with the base sequence described in sequence number 77 or the base sequence of NCBI accession number U64564, preferably DNA with more than 80% homology, and even more preferably DNA with more than 95% homology.

The same applies to the base sequences of mouse MOG [Sequence No. 73, NCBI Accession No. NM_010814], rat MOG [Sequence No. 67, NCBI Accession No. M99485], and cynomolgus monkey MOG [Sequence No. 75, NCBI Accession No. NM_001284856].

As for the functions of MOG, examples include participation in cell adhesion on the myelin sheath and cell surface interactions.

In eukaryotes, the base sequences of proteins-encoding genes often reveal gene polymorphisms. Genes resulting from small-scale mutations in the base sequence due to such polymorphisms are also included in the gene encoding MOG in this invention.

Unless otherwise specified, the homology values in this invention can be values calculated using homology retrieval procedures known to those skilled in the art. For base sequences, values calculated using the default parameters in BLAST [J.Mol.Biol.,215,403(1990)] can be listed, and for amino acid sequences, values calculated using the default parameters in BLAST2 [Nucleic Acids Res.,25,3389(1997); Genome Res.,7,649(1997); http://www.ncbi.nlm.nih.gov/Education/BLASTinfo/information3.htmL] can be listed, etc.

As default parameters, G (cost to open gap) is 5 for base sequences and 11 for amino acid sequences; -E (cost to extend gap) is 2 for base sequences and 1 for amino acid sequences; -q (penalty for nucleotide mismatch) is -3; -r (reward for nucleotide match) is 1; -e (expect value) is 10; -W (wordsize) is 11 residues for base sequences and 3 residues for amino acid sequences; -y [non-vacancy extension decreases]... The threshold, Dropoff(X) for blast extensions in bits, is 20 in the case of blastn and 7 in procedures other than blastn; -X (dropoff threshold for gapped alignment in bits) is 15; -Z (finalX dropoff value for gapped alignment in bits) is 50 in the case of blastn and 25 in procedures other than blastn (http://www.ncbi.nlm.nih.gov/blast/htmL/blastcgihelp.htmL).

Peptides containing partial sequences of the amino acid sequences of the various MOGs described above can be prepared by methods known to those skilled in the art. Specifically, they can be prepared by deleting a portion of the DNA encoding the amino acid sequences of the various MOGs and culturing a transformant containing an expression vector containing the aforementioned DNA. Alternatively, peptides having amino acid sequences obtained by deleting, substituting, or adding one or more amino acids in the amino acid sequences of the various MOGs can be obtained using the same method described above.

In addition, peptides containing amino acid sequences of various MOGs, or peptides having amino acid sequences obtained by deletion, substitution or addition of one or more amino acids in the amino acid sequences of various MOGs, can also be manufactured by chemical synthesis methods such as fluorenyl methoxycarbonyl (Fmoc) method and tert-butyl methoxycarbonyl (tBoc) method.

In this invention, the extracellular region of human MOG refers to the amino acid sequence from position 30 to position 154 or from position 232 to position 247 in the amino acid sequence recorded in sequence number 78 or NCBI accession number AAB08088, preferably the amino acid sequence from position 30 to position 154.

The extracellular region of mouse MOG refers to amino acid positions 30 to 157 or 232 to 247 of the amino acid sequence described in Serial No. 74 or NCBI accession number NP_034944, preferably positions 30 to 157. The extracellular region of rat MOG refers to amino acid positions 28 to 155 or 230 to 245 of the amino acid sequence described in Serial No. 68 or NCBI accession number AAA41628, preferably positions 28 to 155.

The extracellular region of the cynomolgus monkey MOG refers to the amino acid sequence from position 30 to position 154 or from position 232 to position 247 in the amino acid sequence recorded in sequence number 76 or NCBI accession number NP_001271785, preferably the amino acid sequence from position 30 to position 154.

The binding of the antibody of this invention to the extracellular region of MOG can be confirmed by measuring the binding of the antibody of this invention to MOG-expressing cells or recombinant MOG protein using methods such as ELISA, flow cytometry, and surface plasmon resonance assay. Alternatively, it can be confirmed by combining known immunological assays [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Antibodies - A Laboratory Manual, Cold Spring Harbor Laboratory (1988); Monoclonal Antibody Laboratory Manual, Kodansha Scientific (1987)].

The MOG-binding molecules of the present invention are molecules that exhibit brain retention by specifically binding to MOGs in the brain. For example, antibodies are antibodies that exhibit brain retention by binding to MOGs in the brain. Furthermore, the antibodies of the present invention are antibodies that, upon administration to the periphery of an animal, migrate from the periphery across the blood-brain barrier to the brain and bind to MOGs in the brain, thus exhibiting brain retention. The antibodies of the present invention are preferably antibodies with excellent brain retention or antibodies with enhanced brain retention.

In this invention, brain retention refers to the property of the drug remaining in the brain when administered to the test animal. That is, it refers to an increase in the brain concentration (or amount) of the drug, or the presence of a certain concentration at a detectable level, due to at least one of the following: increased migration into the brain, increased accumulation in the brain, decreased migration from the brain to the outside, decreased excretion from the brain to the outside, and decreased decomposition in the brain.

In this invention, "excellent brain retention", "high brain retention" or "improved brain retention" means that when the target substance is administered to the test animal, compared with the control, the concentration (or amount) of the target substance in the brain increases after the same number of days from the date of administration, or the target substance exists in the brain at a certain concentration (amount) that can be detected for a long period of time.

These phenomena are caused by at least one of the following conditions compared to the control: increased migration of the subject into the brain, increased accumulation in the brain, decreased migration from the brain to the outside of the brain, decreased excretion from the brain to the outside of the brain, and decreased decomposition in the brain.

In this invention, examples of excellent brain retention, high brain retention, or improved brain retention include: when the subject is administered to the test animal, the brain concentration (amount) of the subject is higher than that of the control 1 to 10 days after administration, preferably 2 to 10 days, 3 to 10 days after administration, more preferably 4 to 10 days after administration; or the peak value of the brain concentration (or brain amount) of the subject is after the 4th day after administration, preferably after the 5th day, 6th day, 7th day, 8th day, 9th day, more preferably after the 10th day after administration; etc.

Antibodies with good brain retention, high brain retention, or enhanced brain retention can be any type of antibody, as long as they have a higher antibody concentration (antibody amount) in the brain compared to the control antibody, or have the characteristic of being able to persist in the brain for a long time.

Examples of antibodies that, compared to control antibodies, exhibit higher migration and/or accumulation within the brain, lower migration, excretion, and/or degradation within the brain, and higher migration and/or accumulation within the brain than migration, excretion, and/or degradation within the brain, are as follows:

Therefore, examples of antibodies or antibody fragments of the present invention include: antibodies or antibody fragments that, when administered to animals, have a higher antibody concentration (or antibody amount) in the brain after the same number of days from administration compared to a control antibody; or antibodies or antibody fragments that can persist in the brain for a long period of time.

The changes in antibody concentration (or antibody amount) in the brain can be arbitrary. Examples include: the antibody concentration in the brain first reaches a peak during the test and then slowly decreases; the antibody concentration in the brain reaches a peak and then remains at that peak; or the antibody concentration in the brain continues to increase after antibody administration; and so on.

The antibody or antibody fragment of the present invention refers to, for example: an antibody whose concentration or amount in the brain is higher than that of the control antibody on day 4 or day 10 after administration to rats; an antibody whose concentration or amount in the brain is maintained or increased during the period from day 4 to day 10 after administration to rats; or an antibody that can be clearly identified in the brain after day 10 after administration to rats; etc., but is not limited to these.

As a control antibody, any antibody can be used as long as it is of the same species or subclass as the antibody being tested. For example, anti-avermectin (AVM) antibody can be used.

In this invention, the brain can include, but is not limited to, brain parenchyma, ventricles, cerebrospinal fluid, etc.

In this invention, methods for administering antibodies to animals include, but are not limited to, intravenous administration, intraperitoneal administration, subcutaneous administration, intradermal administration, nasal administration, intramedullary administration, etc.

In this invention, methods for determining the brain retention of antibodies include, for example: administering an antibody to an animal and, after several days, recovering the brain tissue, homogenizing it, measuring the antibody concentration in the supernatant after centrifugation, and calculating the amount of antibody per unit brain weight; using the recovered brain tissue to determine the presence of antibodies using known immunological methods; or administering labeled antibodies to an animal and detecting the presence of the antibodies over time using an in vivo imaging system; and so on.

As an antibody of the present invention, one antibody selected from the group consisting of (a) to (q) below can be listed.

(a) The amino acid sequences of CDR1 to 3 of VH are the amino acid sequences recorded in sequence numbers 4, 5 and 6, respectively, and the amino acid sequences of CDR1 to 3 of VL contain the amino acid sequences recorded in sequence numbers 10, 11 and 12, respectively.

(b) The amino acid sequences of CDR1 to 3 of VH are the amino acid sequences described in sequence numbers 16, 17 and 18, respectively, and the amino acid sequences of CDR1 to 3 of VL contain the amino acid sequences described in sequence numbers 22, 23 and 24, respectively.

(c) The amino acid sequences of CDR1 to 3 of VH are the amino acid sequences described in sequence numbers 28, 29 and 30, respectively, and the amino acid sequences of CDR1 to 3 of VL contain the amino acid sequences described in sequence numbers 34, 35 and 36, respectively.

(d) The amino acid sequences of CDR1 to 3 of VHH contain antibody fragments containing the amino acid sequences recorded in sequence numbers 40, 41 and 42, respectively.

(e) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 153, 154 and 155, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 158, 159 and 160, respectively.

(f) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 163, 164 and 165, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 168, 169 and 170, respectively.

(g)Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 173, 174 and 175, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 178, 179 and 180, respectively.

(h)Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 183, 184 and 185, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 188, 189 and 190, respectively.

(i) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 193, 194 and 195, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 198, 199 and 200, respectively.

(j) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 203, 204 and 205, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 208, 209 and 210, respectively.

(k)Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 213, 214 and 215, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 218, 219 and 220, respectively.

(l) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 223, 224 and 225, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 228, 229 and 230, respectively.

(m)Antibodies whose CDR1-3 amino acid sequences of VH contain the amino acid sequences described in sequence numbers 233, 234 and 235, respectively, and whose CDR1-3 amino acid sequences of VL contain the amino acid sequences described in sequence numbers 238, 239 and 240, respectively.

(n) Antibodies in which the amino acid sequences of CDR1-3 of VH contain the amino acid sequences described in sequence numbers 243, 244 and 245, respectively, and the amino acid sequences of CDR1-3 of VL contain the amino acid sequences described in sequence numbers 248, 249 and 250, respectively.

(o) An antibody that competitively binds to MOG with at least one of the antibodies described in (a) to (n) above;

(p) An antibody that binds to an epitope containing any of the antibodies described in (a) to (n) above; and

(q) An antibody that binds to the same epitope as any of the antibodies described in (a) to (n) above.

The antibodies of the present invention include antibodies having amino acid sequences of CDR1-3 of VH and CDR1-3 of VL that show at least 85%, preferably at least 90%, homology with the amino acid sequences of CDR1-3 of VH and VL of any of the antibodies described in (a) to (n) above. Homology of 90% or more is more preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, and 99%.

In this invention, as one type of antibody described in (a) to (n) above, examples include human anti-MOG monoclonal antibodies MOG01, MOG09, MOG14, and alpaca anti-MOG monoclonal VHH antibody iMOG-3Rim1-S32. Furthermore, examples include human chimeric antibodies and humanized antibodies of iMOG-3Rim1-S32.

In this invention, the antibody mentioned above (o) refers to a second antibody that inhibits the binding of the first antibody to MOG when the antibody described in (a) to (n) above is used as the first antibody.

In this invention, the antibody mentioned in (p) refers to a second antibody that binds to a second epitope containing the first epitope, when the antibody described in (a) to (n) is used as the first antibody and the epitope bound by the first antibody is used as the first epitope.

Furthermore, the antibody in (q) of the present invention refers to a second antibody that binds to the first epitope when the antibody described in (a) to (n) above is used as the first antibody and the epitope bound by the first antibody is used as the first epitope.

In addition, as the antibody of the present invention, specifically, one antibody selected from the group consisting of (a) to (n) and (o1) to (o22) below can also be listed.

(a) The amino acid sequence of VH is the amino acid sequence described in sequence number 3, and the amino acid sequence of VL contains the amino acid sequence described in sequence number 9.

(b) The amino acid sequence of VH is the amino acid sequence described in sequence number 15, and the amino acid sequence of VL is an antibody containing the amino acid sequence described in sequence number 21.

(c) The amino acid sequence of VH is the amino acid sequence described in sequence number 27, and the amino acid sequence of VL contains the amino acid sequence described in sequence number 33.

(d) The amino acid sequence of VHH contains an antibody fragment containing the amino acid sequence described in sequence number 39.

(e)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 152 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 157.

(f) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 162 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 167.

(g)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 172 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 177.

(h)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 182 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 187.

(i) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 192 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 197.

(j) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 202 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 207.

(k)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 212 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 217.

(l)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 222 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 227.

(m)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 232 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 237.

(n)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 242 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 247.

(o1)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 252 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 254.

(o2)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 256 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 258.

(o3)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 260 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 262.

(o4)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 264 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 266.

(o5)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 268 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 270.

(o6)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 272 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 274.

(o7)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 276 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 278.

(o8)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 280 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 282.

(o9)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 284 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 286.

(o10)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 288 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 290.

(o11)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 292 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 294.

(o12)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 296 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 298.

(o13)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 300 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 302.

(o14)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 304 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 306.

(o15)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 308 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 310.

(o16)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 312 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 314.

(o17)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 316 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 318.

(o18)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 320 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 322.

(o19)An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 324 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 326.

(o20)Antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 328 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 330.

(o21) An antibody whose amino acid sequence of VH includes the amino acid sequence described in sequence number 332, and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 334; and

(o22)Antibodies whose amino acid sequence of VH includes the amino acid sequence described in sequence number 336 and whose amino acid sequence of VL includes the amino acid sequence described in sequence number 338.

The antibodies of the present invention include antibodies whose VH and VL amino acid sequences show at least 85%, preferably at least 90%, homology with the amino acid sequences of any of the antibodies described in (a) to (n) and (o1) to (o22) above. Homology of 90% or more is more preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, and 99%.

In this invention, as one type of antibody described in (a) to (n) and (o1) to (o22) above, examples include human anti-MOG monoclonal antibodies MOG01, MOG09, and MOG14, and alpaca anti-MOG monoclonal VHH antibody iMOG-3Rim1-S32. Furthermore, examples include iMOG-3Rim1-S32 human chimeric antibody and iMOG-3Rim1-S32 humanized antibody, etc.

In this invention, the EU index refers to the position of amino acid residues shown in the 5th edition (1991) of the Sequences of Proteins of Immunological Interest. Unless otherwise specified, the positions of the amino acid residues shown below all represent the positions of the amino acid residues recorded in the EU index.

Antibody molecules, also known as immunoglobulins (hereinafter referred to as Ig), have a basic structure of a tetramer of polypeptides with two heavy chains (hereinafter referred to as H chains) and two light chains (hereinafter referred to as L chains).

In addition, the H chain is composed of the variable region (also denoted as VH) and the constant region (also denoted as CH) of the H chain starting from the N end, and the L chain is composed of the variable region (also denoted as VL) and the constant region (also denoted as CL) of the L chain starting from the N end.

CH chains are known to have α, δ, ε, γ, and μ chains according to their respective subclasses. In addition, CH chains are composed of CH1, CH2, and CH3 domains starting from the N-terminus.

A domain refers to the functional structural unit of the polypeptide that makes up an antibody molecule. Additionally, the CH2 and CH3 domains are combined and referred to as the Fc region (crystallizable fragment) or simply Fc. CL is known to have _Cλ and _Cκ chains.

Antibodies with CH chains of α, δ, ε, γ, and μ are classified as IgA, IgD, IgE, IgG, and IgM, respectively. Depending on the animal, each antibody subclass may have isotypes. In humans, IgA has isotypes IgA1 and IgA2, and IgG has isotypes IgG1, IgG2, IgG3, and IgG4.

The CH1 domain, hinge domain, CH2 domain, CH3 domain, and Fc region in this invention can be determined using the EU index based on the amino acid residue number starting from the N-terminus.

Specifically, CH1 was identified as the amino acid sequence at EU index positions 118–215, the hinge as the amino acid sequence at EU index positions 216–230, CH2 as the amino acid sequence at EU index positions 231–340, CH3 as the amino acid sequence at EU index positions 341–447, and the Fc region as the amino acid sequence at EU index positions 231–447.

The antibodies used in this invention also include any one of polyclonal antibodies, monoclonal antibodies, and oligoclonal antibodies. Polyclonal antibodies refer to a group of antibody molecules secreted by antibody-producing cells of different clones. Monoclonal antibodies refer to antibodies secreted by antibody-producing cells of a single clone, which recognize a unique epitope (also called an antigenic determinant) and whose amino acid sequences (primary sequences) are all identical. Oligoclonal antibodies refer to a group of antibody molecules obtained by mixing multiple different monoclonal antibodies.

Examples of monoclonal antibodies in this invention include antibodies produced by hybridomas or recombinant antibodies produced by transformants transformed using expression vectors containing antibody genes.

Epitopes can be listed as single amino acid sequences recognized and bound by monoclonal antibodies, three-dimensional structures composed of amino acid sequences, post-translational modified amino acid sequences, and three-dimensional structures composed of post-translational modified amino acid sequences.

Examples of amino acid sequences that have undergone post-translational modification include O-bonded sugar chains with sugar chains bonded to Tyr and Ser with OH substituents, N-bonded sugar chains with sugar chains bonded to Gln and Asn with _NH2 substituents, and amino acid sequences that have been tyrosine-sulfated by sulfate molecules bonded to Tyr with OH substituents.

The epitopes of MOGs bound by the antibodies of this invention can be determined through antibody binding experiments using defective variants obtained by deleting a portion of the MOG domain, mutant variants obtained by replacing a portion of the MOG domain with a domain derived from other proteins, and partial peptide fragments of MOG. Alternatively, antibody binding experiments can be performed using cells expressing the aforementioned defective or mutant variants.

Alternatively, the epitope of the MOG bound by the antibody of the present invention can also be determined by adding the antibody of the present invention to the peptide fragment of the MOG after digestion with a proteolytic enzyme and performing epitope mapping using a known mass spectrometry method.

The antibodies used in this invention also include recombinant antibodies such as mouse antibodies, rat antibodies, hamster antibodies, rabbit antibodies, llama antibodies, camel antibodies, alpaca antibodies, chimeric antibodies, humanized antibodies (also known as "complementarity-determining region (CDR) transplantation antibodies"), and human antibodies.

In this invention, chimeric antibodies refer to antibodies whose VH and VL are derived from different animal species than CH and CL. Antibodies composed of VH and VL from animals other than humans (non-human animals) and CH and CL from human antibodies are called human-type chimeric antibodies. Antibodies composed of VH and VL from animals other than mice and CH and CL from mouse antibodies are called mouse-type chimeric antibodies. Other chimeric antibodies are named in the same way.

As non-human animals, any animal that can produce hybridomas or antibody phage libraries can be used, such as mice, rats, hamsters, rabbits, llamas, camels, alpacas, etc.

Hybridomas are cells that produce monoclonal antibodies with the desired antigen specificity, obtained by fusing B cells obtained by immunizing non-human animals with myeloma cells derived from mice, etc.

An antibody phage library is a library created by cloning the gene of the variable region of an immunoglobulin into a phage, thereby enabling the expression of antigen-binding molecules on its surface. Examples of phages that can be used include M13 phage, and there are no specific limitations.

The antigen-binding molecules displayed on the bacteriophage can be in any form, preferably antibody fragments such as scFv, Fab, and VHH.

In this invention, the antibody phage library can be any one of the following: an immune library, a natural library, or a synthetic library.

An immune library is an antibody-phage library constructed based on antibody genes derived from lymphocytes of animals or patients immunized with antigens. A natural library is an antibody-phage library constructed based on antibody genes derived from lymphocytes of normal animals or healthy humans. A synthetic library is a library obtained by replacing the CDR of the V gene in genomic DNA or a reconstructed functional V gene with oligonucleotides encoding random amino acid sequences of appropriate length.

The following describes the method for preparing human chimeric antibodies. Other chimeric antibodies can also be prepared using the same method.

Human chimeric antibodies can be manufactured by the following method: obtaining cDNA encoding VH and VL from hybridomas derived from non-human animal cells that produce monoclonal antibodies, inserting them into animal cell expression vectors containing DNA encoding CH and CL for human antibodies, respectively, to construct human chimeric antibody expression vectors, and introducing them into animal cells for expression.

Alternatively, it can be manufactured by cloning the genes encoding VH and VL from an antibody phage library derived from non-human animals, inserting them into animal cell expression vectors containing DNA encoding CH and CL for human antibodies, respectively, to construct a human chimeric antibody expression vector, which is then introduced into animal cells for expression.

Humanized antibodies are antibodies obtained by transplanting the amino acid sequences of the CDRs (VH and VL) of non-human animal antibodies into the corresponding CDRs of human antibodies. The regions outside the CDRs of VH and VL are called the frame regions (hereinafter referred to as FRs).

Humanized antibodies can be manufactured by the following method: constructing cDNA encoding the amino acid sequence of VH, which consists of the amino acid sequence of the CDR of a non-human animal antibody and the amino acid sequence of the FR of any human antibody VH, and cDNA encoding the amino acid sequence of VL, which consists of the amino acid sequence of the CDR of a non-human animal antibody and the amino acid sequence of the FR of any human antibody VL, respectively, inserting them into an animal cell expression vector containing DNA encoding the CH and CL of a human antibody, constructing a humanized antibody expression vector, and introducing it into animal cells for expression.

Human antibodies originally refer to antibodies that exist naturally in the human body, but they also include antibodies obtained from human antibody phage libraries and transgenic animals produced from human antibodies.

Human antibodies can be obtained by immunizing mice carrying human immunoglobulin genes with the desired antigen (Tomizuka K. et al., Proc Natl Acad Sci U S A. 97, 722-7, 2000.). Alternatively, human antibodies can be obtained without immunization by selecting human antibodies with the desired binding activity using a phage display library obtained by amplifying antibody genes from human B cells (Winter G. et al., Annu Rev Immunol. 12: 433-55, 1994).

In addition, human B cells can be immortalized by using EB virus to produce cells that generate human antibodies with the desired binding activity, thus obtaining human antibodies (Rosen A. et al., Nature 267, 52-54, 1977).

A human antibody phage library is a library of phages that express antibody fragments such as Fab, scFv, and VHH on their surface by inserting antibody genes prepared from human (healthy or patient) lymphocytes into phage genes. Phages expressing antibody fragments with desired antigen-binding activity can be recovered from this library, using their binding activity against substrates immobilized with antigens as an indicator. These antibody fragments can be further converted into human antibody molecules composed of two complete H chains and two complete L chains using genetic engineering methods.

Human antibody-producing transgenic animals are animals in which human antibody genes are integrated into the chromosomes of a host animal. Specifically, human antibody-producing transgenic animals can be created by introducing human antibody genes into mouse ES cells, transplanting those ES cells into early mouse embryos, and allowing them to develop.

The production of human antibodies from transgenic animals can be carried out by the following method: culturing a hybridoma of human antibodies obtained using the usual hybridoma production method in mammals other than humans, allowing the human antibodies to accumulate in the culture, and purifying the antibody from the culture.

The antibodies of this invention comprise heavy chain antibodies consisting solely of heavy chains. Heavy chain antibodies refer to antibodies obtained from camel-like animals such as llamas, camels, and alpacas, as well as recombinant antibodies made based on such antibodies.

In this invention, an antibody fragment refers to a fragment of an antibody that has antigen-binding activity. Examples include Fab, Fab', F(ab') ₂ , single-chain Fv (scFv), double-chain antibodies, dsFv, peptides containing multiple CDRs, and VHH. Furthermore, in this invention, any antibody fragment containing a partial fragment of an antibody and having MOG-binding activity can include any of the following: antibody fragments obtained by fusing the constant region or a portion of the Fc of an antibody with the antibody fragment, antibody fragments containing a constant region or Fc, etc.

Fab is an antibody fragment with an antigen-binding activity, with a molecular weight of about 50,000, obtained by treating IgG antibodies with papain, a protein-degrading enzyme (the H chain is cleaved at amino acid residue 224). About half of the N-terminal side of the H chain is linked to the entire L chain by a disulfide bond (S-S bond).

F(ab') ₂ is an antibody fragment with an antigen-binding activity, which is slightly larger than the fragment formed by Fab through SS bonds in the hinge region, and has a molecular weight of about 100,000, obtained by treating IgG with pepsin, a protein-degrading enzyme (cleaved at the 234th amino acid residue of the H chain).

Fab' is an antibody fragment with an antigen-binding activity, with a molecular weight of approximately 50,000, obtained by cleaving the SS bond in the hinge region of the aforementioned F(ab') ₂ .

scFv is an antibody fragment with antigen-binding activity obtained by linking a VH to a VL using an appropriate peptide linker (P) such as a linker peptide consisting of any number of linkers (G4S) composed of 4 Gly and 1 Ser residues.

Double-chain antibodies are antibody fragments obtained by forming dimers from scFvs with the same or different antigen binding specificities. They are antibody fragments with divalent antigen-binding activity against the same antigen or specific antigen-binding activity against different antigens.

dsFv refers to a fragment formed by replacing one amino acid residue in VH and VL with a cysteine residue, so that the resulting polypeptide is linked by S-S bonds between the cysteine residues.

A peptide containing a CDR is composed of at least one region of a VH or VL CDR. A peptide containing multiple CDRs may allow the CDRs to bind to each other directly or through appropriate peptide linkers.

Peptides containing CDRs can be manufactured by constructing DNA encoding the CDRs of VH and VL of the antibodies of the present invention, inserting the DNA into a prokaryotic or eukaryotic expression vector, and introducing the expression vector into a prokaryote or eukaryote for expression. Alternatively, peptides containing CDRs can also be manufactured using chemical synthesis methods such as the Fmoc method or the tBoc method.

VHH is the variable region of a heavy chain antibody, also known as a nanobody.

The antibody fragment of the present invention may contain any antibody fragment as long as it is an antibody fragment containing any of the above-mentioned antibody fragments or a portion thereof and having MOG binding activity.

In this invention, an antibody or antibody fragment having one antigen-binding site is referred to as a monovalent antibody. Examples of monovalent antibodies include those described in International Publication Nos. 2014/054804, 2011/090754, 2007/048037, and 2012/116927, which are antibodies or antibody fragments having one antigen-binding site.

In this invention, an antibody molecule or antibody fragment that binds to three or more different antigens or epitopes is referred to as a multispecific antibody. Furthermore, in this invention, an antibody molecule or antibody fragment that binds to two different antigens or epitopes is referred to as a bispecific antibody.

Examples of multispecific or bispecific antibodies include International Publication No. 2009/131239, International Publication No. 2014/054804, International Publication No. 01/077342, U.S. Patent Application Publication No. 2007/0071675, International Publication No. 2007/024715, Wu et al., [Nature Biotechnology, 2007, 25(11), pp. 1290-1297], Labrijn et al., [PNAS 2013, vol. 110, no. 13, pp. 5145-5150], and Jong et al., [http://dx.doi.or The forms described in [g/10.1371/journal.pbio.1002344], Kontermann et al., [mAbs2012, vol.4, issue2, p182-197], Spiess et al., [Molecular Immunology 67(2015)95-106], Ridgway et al., [Protein Engineering, 1996 vol.9 no.7 pp617-621], International Publication No. 2009/080251, International Publication No. 2010/151792 and International Publication No. 2014/033074, etc.

Specifically, the following examples of bispecific antibodies can be listed as examples of bispecific antibodies.

(1) Bispecific antibodies are obtained by applying an amino acid change of S354C/T366W to the CH3 of one heavy chain (heavy chain A) and an amino acid change of Y349C/T366S/L368A/Y407V to the CH3 of the other heavy chain (heavy chain B).

(2) Bispecific antibodies obtained by fusing antibody fragments to the C-terminus of an antibody.

(3) Bispecific antibodies obtained by fusing antibody fragments to the N-terminus of an antibody.

The bispecific antibody described in (1) above can be a bispecific antibody that binds to the antigen-binding site of VH containing heavy chain A and to the MOG, or binds to the antigen-binding site of VH containing heavy chain B and to the antigen present in the brain, or vice versa.

The bispecific antibody described in (2) above can be any one of the following: a bispecific antibody having an antibody fragment bound to the C-terminus of one of the two heavy chains constituting the antibody, or a bispecific antibody having antibody fragments bound to both of the two heavy chains. Furthermore, a suitable linker may exist between the C-terminus of the antibody heavy chain and the antibody fragment.

The antibody fragments of the bispecific antibodies described in (2) above are preferably scFv, Fab and VHH, but are not particularly limited to these.

The bispecific antibody described in (2) above can be a bispecific antibody whose N-terminal antigen-binding site binds to the MOG and whose C-terminal antigen-binding site binds to an antigen present in the brain, or vice versa.

The bispecific antibody described in (3) above refers to a bispecific antibody having an antibody fragment bound to the N-terminus of at least one of the two heavy or light chains constituting the antibody. Furthermore, a suitable linker may exist between the N-terminus of the heavy chain and/or light chain of the antibody and the antibody fragment. The antibody fragments possessed by the bispecific antibody described in (3) above are preferably scFv, Fab, and VHH, but are not particularly limited to these.

In addition, examples of bispecific antibodies described in (3) above include bispecific antibodies having a structure such as _VH1 -CH1- _VH2 -CH1-hinge-CH2-CH3 starting from the N-terminus of the heavy chain, and bispecific antibodies having the above heavy chain structure in which _VH1 and _VH2 respectively form antigen-binding sites with VL. The VL that forms antigen-binding sites with _VH1 and _VH2 can have the same amino acid sequence or different amino acid sequences.

In this invention, the multispecific antibody or bispecific antibody can be any antibody as long as it is a multispecific antibody or bispecific antibody that binds to MOG. Preferably, it is a multispecific antibody or bispecific antibody that binds to both MOG and antigens present in the brain; more preferably, it is a multispecific antibody or bispecific antibody that includes both an antigen-binding site for binding to MOG and an antigen-binding site for binding to antigens present in the brain.

In this invention, antigens present in the brain can include proteins, sugar chains, and lipids, with proteins being preferred.

Examples of proteins found in the brain include MOG, prions, 5T4, AFP, ADAM-10, ADAM-12, ADAM17, AFP, AXL, BSG, C5, C5R, CA9, CA72-4, CCL11, CCL2, CCR1, CCR4, CCR5, CCR6, CD2, CD3E, CD4, CD5, CD6, CD8, CD11, CD18, CD19, CD20, CD22, CD24, CD25, and CD26. 9. CD30, CD32B, CD33, CD37, CD38, CD40, CD40LG, CD44, CD47, CD52, CD55SC1, CD56, CD66E, CD71, CD72, CD74, CD79a , CD79b, CD80, CD86, CD95, CD98, CD137, CD147, CD138, CD168, CD200, CD248, CD254, CD257, CDH3, CEA, CEACAM1, CEA CAM5, CEACAM6, CEACAM8, Claudin3, Claudin4, c-Met, CS-1, CSF2RA, CSPG-4, CTLA4, CRF-1, Cripto, CXCR4, CXCR5, DLL4, DR4, DR5, ED-B, EFNA2, EGFR, EGFRvIII, ETBR, ENPP3, EPCAM, EphA2, ERB B2, ERBB3, ERBB4, FAPα, FAS, FcγRI, FCER2, FGFR1, FGFR2, FGFR3, FGFR4, FLT1, FOLH1, FOLR1, GDF2, GFR, GLP1R, glypican-3, GPNMB, GRP78, HB-EGF, HGF, HLA-DRβ, ICAM1, IFNA1, IFNA1, IgE, IgE-Fc, IGF1R, IL10, IL 12B, IL13, IL15, IL17A, IL1A, IL1B, IL2RA, IL4, IL5, IL5RA, IL6, IL6R, IL9, IL2Rα, IL2Rβ, IL2Rγ, INSR, ITGA2, IT GA2B2, ITGB3, ITGA4, ITGB7, ITGA5, ITGAL, ITGAV, ITGB3, ITGB2, KDR, L1CAM, mesothelin, MMP14, MMP15, MS T1R, MSTN, MUC1, MUC4, MUC16, MUC5AC, myostatin, NECTIN4, NGF, NOTCH, NRG1, NRP, OX40, OX40L, PDGFA, PDGFB, PDGFRA, PDGFRB, PD1, PDL1, PSCA, SLAM7, SLC44A4, TAG-72, TCR, TGFB1, TGFB2, TGFBR, TNF, TNFR, TNFRSF10 A, TNFRSF10B, TNFRSF12A, TNFSF13, TNFSF14, TNFSF2, TNFSF7, TRAILR2, TRKA, TRKB, TRKC, VEGF, VEGFR, VLA-4, CGRP, alpha-synuclein, TDP-43, Tau, FUS, amyloid-beta (Aβ), APP, BACE1, presenilin, LINGO-1, Nogo, polyglutamine (polyQ), androgen receptor, huntingtin, spinocerebellar ataxia type I protein (ataxin 1), spinocerebellar ataxia type II protein (ataxin 2), RGMA, phosphorylated Tau (Phospho-Tau) or phosphorylated alpha-synuclein, etc., but not limited to these proteins.

Examples of sugar chains that exist in the brain include Lewis-x, Lewis-y, or CD15, but are not limited to these sugar chains.

Lipids present in the brain include, but are not limited to, GD1a, GD2, GD3, GM1, GM2, GM3, or phosphatidylserine.

The antibody or antibody fragment of the present invention also includes an antibody containing any amino acid that has been post-translational modified. Examples of post-translational modifications include, for example, the deletion of lysine residues at the C-terminus of the H chain [lysine clipping], and the conversion of glutamine residues at the N-terminus of the polypeptide to puroglutamine [Beck et al, Analytical Chemistry, 85, 715-736 (2013)].

The antibody or antibody fragment of the present invention can undergo amino acid alteration in the Fc region. Examples of amino acid alterations in the Fc region include, for example, alterations for stabilizing the antibody or for controlling the half-life in the blood. Specifically, examples of amino acid alterations in the Fc region include International Publication Nos. 2006/033386, 2006/075668, 2011/122011, and 2009/125825.

The antibodies or antibody fragments of the present invention also comprise fusion antibodies or fusion antibody fragments modified from the antibodies or antibody fragments. The method of modifying the antibody is not particularly limited; any method can be used as long as the desired amino acid residues and sugar chains can be modified.

Examples of such modifications include chemical modifications using chemical reactions [Introduction to Antibody Engineering, Diren Book House (1994); Kolbet al., Angew Chem Int Ed Engl. 40. 2004-21, 2001], and modifications using gene engineering methods that introduce recombinant protein expression vectors into appropriate host cells for expression.

In this invention, the molecules used to modify antibodies or antibody fragments can include, for example, hydrophilic polymers, amphiphilic polymers, and functional molecules. Examples of the aforementioned hydrophilic and amphiphilic polymers include, for example, polyoxyethylene, molecules containing polyols or polysaccharides.

Examples of polyoxyethylenes include polyethylene glycol (PEG), polypropylene glycol, and polypropylene glycol, which are composed of straight or branched chains.

Examples of molecules containing polyols or polysaccharides include linear starch, dextran, pullulan, or glycogen, which are formed from linear or branched polyglycerols.

The molecular weight of the molecules containing hydrophilic or amphiphilic polymers is not particularly limited, but is preferably 100 Da or more, for example, 100 Da to 100 kDa.

Examples of functional molecules include antigen-binding molecules and their fragments, drugs, physiologically active peptides, physiologically active proteins, nucleic acids, radiolabeled compounds, glycans, lipids, or fluorescent compounds. The result of modifying antigen-binding molecules with functional molecules is a bispecific antibody, a molecule possessing dual specificity.

Examples of antigen-binding molecules include antibodies, receptors, and ligands.

As a fragment of an antigen-binding molecule, any fragment can be used, as long as it is a fragment of the aforementioned antigen-binding molecule and has antigen-binding activity.

As drugs, examples include alkylating agents, nitrosoureas, metabolic antagonists, antiviral agents, antibiotics, plant alkaloids, topoisomerase inhibitors, tubulin polymerization inhibitors, hormone therapy agents, hormone antagonists, aromatase inhibitors, P-glycoprotein inhibitors, platinum complex derivatives, M-phase inhibitors or kinase inhibitors, etc. [Clinical Oncology, Cancer and Chemotherapy Publishing House (1996)], steroids such as hydrocortisone and prednisolone, nonsteroidal agents such as aspirin and indomethacin, immunomodulators such as gold thiomalate and penicillamine, immunosuppressants such as cyclophosphamide and azathioprine, or antihistamines such as chlorpheniramine maleate or chlormastine, etc. [Inflammation and Anti-inflammatory Therapy, Ichi-Den Publishing Co., Ltd. (1982)], etc.

Examples of anticancer agents include mertansine, emtansine, amifostine, cisplatin, dacarbazine (DTIC), styromycin, dichloromethyldiethylamine (nitrogen mustard), streptozotocin, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), doxorubicin (doxorubicin), epirubicin, gemcitabine (Gemzal), daunorubicin, procarbazine, mitomycin, cytarabine, etoposide, 5-fluorouracil, fluorouracil, vincristine, bleomycin, donomycin, pelocyticus, estradiol, paclitaxel, docetaxel, and interleukin. Asparaginase, Busulfan, Carboplatin, Oxaliplatin, Nedaplatin, Cladribine, Camptothecin, 10-Hydroxy-7-ethylcamptothecin (SN38), Fluorouracil, Fludarabine, Hydroxyurea, Idarubicin, Mesna, Irinotecan (CPT-11), Topotecan, Mitoxantrone, Topotecan, Leuprolide, Medroxyprogesterone acetate, Melphalan, Mercaptopurine, Hydroxyurea, Procainamide, Mitotane, Pegaspargase, Pentostatin, Piperbromide, Streptozomycin, Tamoxifen, Goserelin, Leuprorelin, Flutamide, Teniposide, Testrolide, Thioguanine, Thiotepa, Uramustine, Vinorelbine, Chlormethazine, Hydrocortisone Prednisolone, methylprednisolone, vindesine, nimustine, semustine, capecitabine, raltitrexed, azacitidine, UFT, oxaliplatin, gefitinib (Iressa), imatinib (STI571), erlotinib, FMS-like tyrosine kinase 3 (Flt3) inhibitors, vascular endothelial growth factor receptor (VEGFR) inhibitors, fibroblast growth factor receptor (FGFR) inhibitors, epidermal growth factor receptor (EGFR) inhibitors such as erlotinib, rhizobactam, 17-allylamino-17-demethoxygerdomyl, rapamycin, acridine, all-trans retinoic acid, thalidomide, Lenalidomide, anastrozole, fastrozole, letrozole, exemestane, gold thiomalate, D-penicillamine, benzylamine, azathioprine, imidazolidin, cyclosporine, rapamycin, hydrocortisone, bisalostine (tagresin), tamoxifen, thiamethoxam, progestins, estrogens, anastrozole (Renin), lipoprotein, aspirin, indomethacin, celecoxib, azathioprine, penicillamine, gold thiomalate, chlorpheniramine maleate, chlorpheniramine, chlormastine, retinoic acid, bisalostine, arsenic, bortezomib, allopurinol, cazithromycin, teimomab, tagresin, oxazomicin, clarithromycin, levofloxacin, ketoconazole, aminoglutethimide, suramin, methotrexate, maytanol or their derivatives, etc.

In addition to the methods described above, other methods for binding a drug to an antibody or an antibody fragment include binding the drug to the amino group of the antibody using glutaraldehyde, or binding the amino group of the drug to the carboxyl group of the antibody using water-soluble carbodiimide.

Physiologically active peptides or proteins include, for example, interferon (hereinafter referred to as IFN)-α, IFN-β, IFN-γ, interleukin (hereinafter referred to as IL)-2, IL-12, IL-15, IL-18, IL-21, IL-23, cytokines or proliferation factors that activate immune cells such as NK cells, macrophages or neutrophils, such as granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF); protein-degrading enzymes such as hydrolases, lyases and isomerases; enzymes such as acid sphingomyelinase; toxins such as ricin, diphtheria toxin or ONTAK, bacterial toxins and plant toxins; antimicrobial peptides with cell membrane toxicity; peptides with cell membrane binding or cell membrane permeability and their derivatives.

As a nucleic acid, it can be any molecule as long as it is a nucleotide or a molecule that has the same function as that nucleotide. Examples include siRNA, microRNA, antisense RNA, DNA aptamers, etc.

As radiolabeled compounds, any nucleus used for diagnostic or therapeutic purposes is acceptable. Examples include ^3H , ^14C , ^32P , ^33P , 35S, ^51Cr , ^57CO , ^18F , ^153Gd , 159Gd, ^64Cu , ^68Ge , ^166Ho , ^115In , ^113In , ^112In , ^111In , ^131I , ^125I , ^123I , ^121I , ^140La , ^177Lu , ^54Mn , ^99Mo , ^103Pd , ^142Pr , ^149Pm , ^186Re , ^188Re , ^211At , ^105Rh , ^97Ru , ^153Sm , ^{47Sc , 75Se} , ^85Sr , ^{99Tc , 201} Ti, ¹¹³ Sn, ¹¹⁷ Sn, ¹³³ Xe, ¹⁶⁹ Yb, ¹⁷⁵ Yb, ⁹⁰ Y and ⁶⁵ Zn, or compounds containing the above nuclei.

Radiolabeled compounds can be directly bound to antibodies using methods such as the chloramine-T method. Alternatively, substances that chelate radiolabeled compounds can be bound to antibodies. Examples of chelating agents include DOTA, PA-DOTA, TRITA, and DTPA. Antibodies modified with chelating agents and modified antibodies labeled with radiolabeled compounds using chelating agents are also included in the antibodies of this invention.

Examples of sugar chains include monosaccharides, disaccharides, or oligosaccharides such as fucose, mannose, glucose, allose, aldose, gulose, idose, galactose, tarose, ribose, arabinose, xylose, lysose, erythose, erythrose, threose, cellobiose, maltose, isomaltose, lactose, lipoarabinomannan, Lewis X trisaccharides, and sialic acid Lewis X tetrasaccharides. Additionally, natural substances containing sugar chains known as immune adjuvants can also be included, such as β(1→3)glucan (lentinan, cizolose) or α-galactosylceramide (KRN7000).

Examples of lipids include simple lipids (neutral lipids) that are esters of fatty acids and various alcohols and their analogues. Examples include oils (e.g., triacylglycerols), waxes (e.g., fatty acid esters of higher fatty alcohols); complex lipids such as sterol esters, cholesterol esters, and fatty acid esters of vitamins, which have polar groups such as phosphate, sugar, sulfate, and amine in addition to fatty acids and alcohols, such as phospholipids (e.g., glycerophospholipids and sphingomyelins) and glycolipids (e.g., glyceroglycolipids and sphingolipids); and derivative lipids that are fat-soluble and derived from compounds produced by the hydrolysis of simple lipids and complex lipids, such as fatty acids, higher fatty alcohols, fat-soluble vitamins, steroids, and hydrocarbons.

Examples of fluorescent compounds include fluorescein series such as FITC, rhodamine series, Cy3, Cy5, eosin series, Alexa Fluor series and NBD series, luminescent substances such as acridinium esters or rofin, and fluorescent proteins such as green fluorescent protein (GFP).

The antibody or antibody fragment of the present invention can bind directly or via a suitable linker to the aforementioned hydrophilic or amphiphilic polymers and functional molecules. Examples of linkers include esters, disulfides, hydrazones, and dipeptides.

When using genetic engineering methods to modify the antibody or antibody fragment of the present invention to produce a fusion antibody or fusion antibody fragment, a cDNA encoding a protein can be linked to the cDNA encoding the antibody to construct DNA encoding the fusion antibody or fusion antibody fragment. This DNA can be inserted into a prokaryotic or eukaryotic expression vector, and the expression vector can be introduced into a prokaryotic or eukaryotic organism to express the fusion antibody or fusion antibody fragment, thereby producing the fusion antibody or fusion antibody fragment.

The compositions of the present invention can be any composition as long as they contain the antibody or antibody fragment of the present invention. In addition to containing the antibody or antibody fragment, the compositions may also contain suitable carriers, stabilizers, or other additives.

Examples of compositions for detection or assay include, for example, compositions containing the antibody or antibody fragment of the present invention. Examples of compositions for formulation include, for example, pharmaceutical compositions (therapeutic agents) containing the antibody or antibody fragment of the present invention as active ingredients, which are formulated together with a pharmacologically acceptable carrier into a desired dosage form.

In this invention, the composition used for detection or assay can be any composition as long as it contains the antibody or antibody fragment of this invention and is capable of detecting or assaying the antigen specifically bound by the antibody or antibody fragment of this invention. Examples of antigens specifically bound by the antibody or antibody fragment of this invention include MOG, or MOG and antigens present in the brain.

The antibody or antibody fragment of the present invention has the property of binding to and remaining in the brain of MOGs when administered to animals. Therefore, by using a detection or assay composition containing the antibody or antibody fragment, it is possible to maintain or increase the antibody concentration in the brain, and to detect or measure MOGs, or MOGs and antigens present in the brain, for extended periods, and/or to detect or measure MOGs, or MOGs and antigens present in the brain, with high sensitivity.

For example, when the composition for detection or assay is a composition containing a bispecific antibody that binds to MOG and antigens present in the brain, it is possible to detect or assay the MOG bound by the bispecific antibody and antigens present in the brain for a long time; and/or to detect or assay the MOG and antigens present in the brain with high sensitivity.

In addition, for example, when the composition for detection or determination is a composition containing a fusion antibody or fusion antibody fragment labeled with a radiolabeled compound or fluorescent dye that binds to MOG, MOG can be detected or determined for a long time; and/or MOG can be detected or determined with high sensitivity.

The pharmaceutical composition (therapeutic agent) containing the antibody of the present invention can be a therapeutic agent for any disease expressing the antibody of the present invention or an antigen specifically bound by the antibody fragment, preferably a therapeutic agent for brain diseases.

Examples of brain diseases include Alzheimer's disease, Alzheimer's disease in its early stages, Huntington's disease, Parkinson's disease, brain tumors, multiple sclerosis, muscular dystrophy, amyotrophic lateral sclerosis, multiple system atrophy, progressive supranuclear palsy, nigrostriatal degeneration, olivary pontocerebellar atrophy, spinobulbar muscular atrophy, spinocerebellar degeneration, cerebrovascular diseases, epilepsy, migraine, ADHD, Creutzfeldt-Jakob disease, corticobasal ganglia degeneration, lysosomal storage diseases, depression, and dystonia.

The antibody of the present invention has the property of binding to MOGs in the brain and remaining in the brain when administered to animals. Therefore, by using a therapeutic agent containing this antibody or antibody fragment, the antibody or antibody fragment can be maintained in the brain for a long time, the antibody concentration in the brain can be increased, and a therapeutic effect can be shown on the aforementioned diseases.

For example, when the therapeutic agent is a bispecific antibody that binds to both the MOG and an antigen present in the brain, it can show therapeutic effects on brain diseases associated with the antigen present in the brain that the bispecific antibody binds to.

Furthermore, for example, when the therapeutic agent is a fusion antibody or fusion antibody fragment modified with a low-molecular-weight agent that binds to an MOG, the low-molecular-weight agent can exhibit a therapeutic effect on the targeted brain disease. In this case, the therapeutic effect is higher when using the therapeutic agent of the present invention compared to using the low-molecular-weight agent alone, and is therefore preferred.

Therapeutic agents containing the antibodies or antibody fragments of the present invention may contain only the antibody or antibody fragment as an active ingredient, but are generally preferably provided as pharmaceutical preparations mixed with one or more pharmacologically acceptable carriers and manufactured using any method known in the field of pharmaceutical formulation.

The preferred route of administration is the one most effective for treatment. Examples include oral administration or non-oral administration such as oral, intraoral, intrarespiratory, rectal, subcutaneous, intradermal, intramuscular, intravenous, intramuscular, intracerebral, intramedullary, intranasal, intraperitoneal, or intravenous administration. Intravenous or intracerebral administration is particularly preferred. As for the form of administration, examples include sprays, capsules, tablets, powders, granules, syrups, emulsions, suppositories, injections, ointments, or patches.

Dosage or frequency of administration varies depending on the target therapeutic effect, method of administration, duration of treatment, age, and weight. Generally, the dosage for adults is 10 μg/kg to 20 mg/kg per day.

In addition, the present invention also includes a method for retaining an antibody in the brain using the antibody or antibody fragment of the present invention, a method for improving the brain retention of antibodies, and a method for increasing the antibody concentration (or amount of antibodies) in the brain.

In addition, the present invention relates to peptides that bind to MOGs, nucleic acids containing a base sequence encoding the peptide, transformed cells containing a carrier containing the nucleic acid, a method for manufacturing the peptide including culturing the transformed cells and collecting the peptide from the culture medium, compositions containing the peptide, or methods for detecting or measuring antigens present in the brain using the peptide or the composition, methods for diagnosing or treating brain diseases, methods for improving the brain retention of peptides, or methods for increasing the amount of peptides in the brain.

The peptides of the present invention comprise fusion peptides obtained by modifying peptides.

Unless otherwise specified, the definitions of various terms related to peptides that bind to MOG shall be the same as those used in the definitions of terms described above for antibodies that bind to MOG.

The following describes in detail the manufacturing method of the antibody or antibody fragment of the present invention, the treatment method for the disease, and the diagnostic method for the disease.

1. Antibody manufacturing methods

(1) Preparation of antigen

MOG, as an antigen, or MOG-expressing cells, can be obtained by introducing an expression vector containing cDNA encoding the full-length or partial length of MOG into E. coli, yeast, insect cells, or animal cells. Alternatively, MOG can be obtained by purifying it from various animal cell lines, animal cells, and animal tissues that express MOG in large quantities.

Alternatively, animal cell lines, animal cells, and animal tissues can be used directly as antigens. Furthermore, synthetic peptides containing partial MOG sequences can be prepared using chemical synthesis methods such as the Fmoc or tBoc methods and used as antigens.

Known tags such as FLAG or His can be appended to the C-terminus or N-terminus of a synthetic peptide containing a portion of the MOG sequence.

The MOG used in this invention can be manufactured by expressing the DNA encoding the MOG in a host cell using methods described in, for example, Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989) or Current Protocols in Molecular Biology, John Willie & Son Press (1987-1997).

First, a recombinant vector is prepared by inserting a full-length cDNA containing the portion encoding MOG downstream of the promoter of a suitable expression vector. Alternatively, a DNA fragment of appropriate length containing the portion encoding a polypeptide, prepared based on the full-length cDNA, can be used. Next, the resulting recombinant vector is introduced into a host cell suitable for the expression vector, thereby obtaining a transformant that produces the polypeptide.

As an expression vector, any expression vector that can autonomously replicate in the host cell or integrate into the chromosome and contains an appropriate promoter at the site where the DNA encoding the polypeptide can be transcribed is acceptable. As for the host cell, any host cell capable of expressing the target gene, such as microorganisms like Escherichia coli, yeast, insect cells, or animal cells, is acceptable.

When using prokaryotes such as *Escherichia coli* as host cells, the expression vector is preferably one that can autonomously replicate in prokaryotes and contains a promoter, a ribosome-binding sequence, DNA encoding a portion of the human MOG, and a transcription termination sequence. Furthermore, the expression vector does not necessarily need to contain a transcription termination sequence, but it is preferable to place the transcription termination sequence immediately downstream of the structural gene. In addition, the recombinant vector may contain a gene controlling the promoter.

As the expression vector, a plasmid is preferably used that adjusts the distance between the Schindalgano sequence (also known as the SD sequence), which serves as the ribosome binding sequence, and the start codon to an appropriate distance (e.g., 6 to 18 bases).

In addition, the base sequence of the DNA encoding MOGs can be substituted to form codons that are best suited for expression within the host, thereby improving the productivity of the target MOGs.

As expression vectors, any vector that can function in the host cells can be used. Examples include pBTrp2, pBTac1, pBTac2 (manufactured by Roche Diagnostics), pKK233-2 (manufactured by Pharmacia), pSE280 (manufactured by Invitrogen), pGEMEX-1 (manufactured by Prometheus), pQE-8 (manufactured by Qiagen), pKYP10 (Japanese Patent Application Publication No. 58-110600), pKYP200 [Agricultural Biological Chemistry, 48, 669 (1984)], pLSA1 [Agric. Biol. Chem., 53, 277 (1989)], pGEL1 [Proc. Natl. Acad. Sci. USA, 82, 4306 (1985)], and pBluescript II SK(-) (manufactured by Stratagene). pTrs30 [prepared from Escherichia coli JM109/pTrS30 (FERM BP-5407)], pTrs32 [prepared from Escherichia coli JM109/pTrS32 (FERM BP-5408)], pGHA2 [prepared from Escherichia coli IGHA2 (FERM BP-400), Japanese Patent Application Publication No. 60-221091], pGKA2 [prepared from Escherichia coli IGKA2 (FERM BP-6798), Japanese Patent Application Publication No. 60-221091] -221091 Publication No. [, pTerm2 (US Patent No. 4,686,191, US Patent No. 4,939,094, US Patent No. 160,735), pSupex, pUB110, pTP5, pC194, pEG400 [J. Bacteriol., 172, 2392 (1990)], pGEX (manufactured by Pharmacia), pET system (manufactured by Novagen), or pME18SFL3, etc.

As a promoter, any promoter can be used as long as it can function in the host cell it is used in. Examples include promoters derived from E. coli or bacteriophages, such as the trp promoter (Ptrp), lac promoter, PL promoter, PR promoter, or T7 promoter. In addition, examples include artificially designed promoters such as tandem promoters (two Ptrp promoters in series), tac promoters, lacT7 promoters, or let I promoters.

Examples of host cells include Escherichia coli XL1-Blue, Escherichia coli XL2-Blue, Escherichia coli DH1, Escherichia coli MC1000, Escherichia coli KY3276, Escherichia coli W1485, Escherichia coli JM109, Escherichia coli HB101, Escherichia coli No.49, Escherichia coli W3110, Escherichia coli NY49, and Escherichia coli DH5α.

As a method for introducing recombinant vectors into host cells, any method that introduces DNA into the host cells in question can be used, such as the method using calcium ions [Proc.Natl.Acad.Sci.USA,69,2110(1972); Gene,17,107(1982); Molecular & General Genetics,168,111(1979)].

When using animal cells as a host, any expression vector that can function within animal cells can be used as an expression vector. Examples include pcDNAI, pCDM8 (manufactured by Funakoshi), pAGE107 [Japanese Patent Application Publication No. 3-22979; Cytotechnology, 3, 133 (1990)], pAS3-3 (Japanese Patent Application Publication No. 2-227075), pCDM8 [Nature, 329, 840 (1987)], pcDNAI/Amp (manufactured by Invitrogen), and pcD... NA3.1 (manufactured by Invitrogen), pREP4 (manufactured by Invitrogen), pAGE103 [J. Biochemistry, 101, 1307 (1987)], pAGE210, pME18SFL3, pKANTEX93 (International Publication No. 97/10354), N5KG1val (US Patent No. 6001358), INPEP4 (manufactured by Biogen-IDEC), pCI (manufactured by Promega), and transposon vectors (International Publication No. 2010/143698), etc.

As a promoter, any promoter capable of functioning in animal cells can be used. Examples include the promoter of the immediate early (IE) gene of cytomegalovirus (CMV), the early promoter of SV40, promoters of retroviruses, metallothionein promoters, heat shock promoters, SRα promoters, or promoters or enhancers of Moloney's mouse leukemia virus. Alternatively, the enhancer of the IE gene of human CMV can be used in conjunction with the promoter.

Examples of host cells include human leukemia cells (Namalwa cells), monkey COS cells, and Chinese hamster ovary cells (CHO cells) [Journal of Experimental Medicine, 108, 945 (1958); Proc. Natl. Acad. Sci. USA, 60, 1275 (1968); Genetics, 55, 513 (1968); Chromosom a,41,129(1973);Methods in Cell Science,18,115(1996); Radiation Research,148,260(1997);P roc.Natl.Acad.Sci.USA,77,4216(1980); Proc.Natl.Acad.Sci.,60,1275(1968); Cell,6,121(1975) [Molecular Cell Genetics, Appendix I, II (pp. 883-900)]; CHO cells (CHO/DG44 cells) deficient in dihydrofolate reductase gene (hereinafter referred to as dhfr) [Proc. Natl. Acad. Sci. USA, 77, 4216 (1980)], CHO-K1 (ATCC CCL-61), DUkXB11 (ATCC CCL-9096) Pro-5 (ATCC CCL-1781), CHO-S (Life Technologies, Cat#11619), Pro-3, rat myeloma cells YB2/3HL.P2.G11.16Ag.20 (or also known as YB2/0), mouse myeloma cells NS0, mouse myeloma cells SP2/0-Ag14, Syrian hamster cells BHK or HBT5637 (Japanese Patent Application Publication No. 63-000299), etc.

As a method for introducing expression vectors into host cells, any method that introduces DNA into animal cells can be used, such as electroporation [Cytotechnology, 3, 133 (1990)], calcium phosphate method (Japanese Patent Application Publication No. 2-227075) or liposome transfection method [Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)].

The transformants obtained above, derived from microbial or animal cells containing an expression vector integrating DNA encoding MOG, are cultured in a culture medium to allow the MOG to be generated and accumulated in the culture medium. The MOG is then collected from the culture medium, thereby enabling the production of MOG. The method for culturing the transformants in the culture medium can be performed according to the methods commonly used in host culture.

When expressed in cells derived from eukaryotes, MOGs with attached sugars or sugar chains can be obtained.

When culturing microorganisms transformed using expression vectors employing inducible promoters, inducers can be added to the culture medium as needed. For example, isopropyl-β-D-thiogalactopyranoside can be added to the culture medium when culturing microorganisms transformed using expression vectors employing trp promoters, indoleacrylic acid can be added.

Examples of culture media for transformants obtained from animal cells include commonly used RPMI 1640 medium [The Journal of the American Medical Association, 199, 519 (1967)], Eagle's MEM medium [Science, 122, 501 (1952)], Duchenne's modified MEM medium [Virology, 8, 396 (1959)], 199 medium [Proc. Soc. Exp. Biol. Med., 73, 1 (1950)], or Iscove's modified Duchenne MEM medium (IMDM), or media obtained by adding fetal bovine serum (FBS) to these media. Culture is typically carried out for 1–7 days under conditions of pH 6–8, 30–40°C, and 5% _CO2 . Additionally, antibiotics such as kanamycin and penicillin can be added to the culture medium as needed.

In addition to direct expression, other methods for expressing genes encoding MOG include secretory production or fusion protein expression [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989)].

Methods for producing MOGs include, for example, methods of production within host cells, methods of secretion outside host cells, or methods of production on the outer membrane of host cells. The appropriate method can be selected by changing the host cell used or the structure of the MOG produced.

When MOGs are produced inside or on the host cell membrane, they can be actively secreted outside the host cell using methods described in Paulson et al. [J. Biol. Chem., 264, 17619 (1989)], Luo et al. [Proc. Natl. Acad. Sci., USA, 86, 8227 (1989); Genes Develop., 4, 1288 (1990)], Japanese Patent Application Publication No. 05-336963, or International Patent Application Publication No. 94/23021. Alternatively, MOG production can be increased using gene amplification systems employing genes such as dihydrofolate reductase (Japanese Patent Application Publication No. 2-227075).

The obtained MOGs can be separated and purified as follows: When the MOGs are expressed in a soluble state within the cells, the cells are recovered by centrifugation after culture, suspended in an aqueous buffer, and then the cells are disrupted using an ultrasonic disruptor, French press, Manton-Gaulin homogenizer, or Dieno mill to obtain a cell-free extract. The purified product can be obtained from the supernatant obtained by centrifuging the cell-free extract using conventional protein separation and purification methods, such as solvent extraction alone or in combination, salting out with ammonium sulfate, desalting, precipitation with organic solvents, anion exchange chromatography using resins such as DEAE-agarose and DIAION HPA-75 (manufactured by Mitsubishi Chemical Corporation), cation exchange chromatography using resins such as S-agarose FF (manufactured by Pharmacia Corporation), hydrophobic chromatography using resins such as butyl agarose and phenyl agarose, gel filtration using molecular sieves, affinity chromatography, focusing chromatography, or electrophoresis such as isoelectric point electrophoresis.

When MOGs are expressed intracellularly as insoluble forms, the cells are recovered and lysed in the same manner as described above, followed by centrifugation to recover the insoluble MOGs as a precipitate fraction. The recovered insoluble MOGs are then soluble using a protein denaturing agent. The soluble solution is diluted or dialyzed to restore the MOGs to their normal stereostructure. The purified peptide product can then be obtained using the same separation and purification method as described above.

When MOG or its glycomodified derivatives are secreted extracellularly, the MOG or its glycomodified derivatives can be recovered from the culture supernatant. The culture is then processed using methods such as centrifugation as described above to obtain a soluble fraction. The purified product can then be obtained from this soluble fraction using the same separation and purification method as described above.

Alternatively, the MOG used in this invention can also be manufactured using chemical synthesis methods such as the Fmoc method or the tBoc method. Furthermore, it can be chemically synthesized using peptide synthesizers manufactured by Advanced ChemTech, PerkinElmer, Pharmacia, Protein Technology Instruments, Synthecell-Vega, Perceptive, or Shimadzu Corporation.

(2) Animal immunization and preparation of antibody-generating cells for fusion

Immunize mice, rats, rabbits, or hamsters aged 3 to 20 weeks with the antigen obtained in (1), and collect antibody-producing cells from the spleen, lymph nodes, and peripheral blood of the animals. Alternatively, llamas, alpacas, camels, and other animals can also be used as immunized animals.

Immunization is performed by administering the antigen, along with an appropriate adjuvant such as complete Freund's adjuvant or aluminum hydroxide gel with pertussis vaccine, subcutaneously, intravenously, or intraperitoneally to the animal. In the case of a partial peptide antigen, it is conjugated with a carrier protein such as BSA (bovine serum albumin) or keyhole limpet hemocyanin (KLH) to serve as an immunogen.

When immunizing mice or rats, the antigen is administered 5 to 10 times every 1 to 2 weeks after the first administration. Blood is collected from the fundus venous plexus on days 3 to 7 after each administration, and the antibody titer of the serum is determined using enzyme-linked immunosorbent assay (ELISA) [Antibodies-A Laboratory Manual, Cold Spring Harbor Laboratory (1988)]. Animals whose serum shows sufficient antibody titer against the antigen used for immunization are used as the source of antibody-producing cells for fusion.

Three to seven days after the final administration of the antigen, the spleen and other tissues containing antibody-producing cells were removed from the immunized animals to collect antibody-producing cells. If spleen cells were used, the spleen was finely chopped, dispersed, and centrifuged to remove red blood cells, thereby obtaining antibody-producing cells for fusion.

The same method is used to immunize other immunized animals, which can produce antibody-producing cells. The immunization interval and the period from the last immunization to tissue removal can be selected according to the species of the immunized animals.

(3) Preparation of myeloma cells

As myeloma cells, cell lines derived from mice were used, such as the 8-azaguanine resistant mouse (derived from BALB/c) myeloma cell lines P3-X63Ag8-U1 (P3-U1) [Current Topics in Microbiology and Immunology, 18, 1 (1978)], P3-NS1/1-Ag41 (NS-1) [European J. Immunology, 6, 511 (1976)], SP2/0-Ag14 (SP-2) [Nature, 276, 269 (1978)], P3-X63-Ag8653 (653) [J. Immunology, 123, 1548 (1979)], or P3-X63-Ag8 (X63) [Nature, 256, 495 (1975)].

The myeloma cells were passaged in normal culture medium [RPMI 1640 medium supplemented with glutamine, 2-mercaptoethanol, gentamicin, FBS and 8-azaguanine], and passaged into normal culture medium 3-4 days before cell fusion, ensuring a cell count of more than 2 × ^10⁷ on the day of fusion.

(4) Preparation of hybridomas from cell fusion and monoclonal antibody generation

The fusion antibody-generated cells obtained in (2) and the myeloma cells obtained in (3) were thoroughly washed with minimum essential medium (MEM) or PBS (1.83 g disodium hydrogen phosphate, 0.21 g potassium dihydrogen phosphate, 7.65 g sodium chloride, 1 liter of distilled water, pH 7.2). The cells were mixed in a ratio of fusion antibody-generated cells to myeloma cells of 5 to 10:1. After centrifugation, the supernatant was removed.

After thoroughly dispersing the precipitated cell population, a mixture of polyethylene glycol-1000 (PEG-1000), MEM medium, and dimethyl sulfoxide was added while stirring at 37°C. Further, 1-2 mL of MEM medium was added every 1-2 minutes, repeating this process several times until the total volume reached 50 mL.

After centrifugation, remove the supernatant. Gently disperse the precipitated cell population and then gently suspend the cells in HAT medium (normal medium supplemented with hypoxanthine, thymidine, and aminopterin). Incubate the suspension at 37°C for 7–14 days in a 5% _CO2 incubator.

After culturing, a portion of the culture supernatant was extracted, and hybridoma selection methods, such as binding analysis described later, were used to select cell populations that reacted with MOG antigens but not with non-MOG antigens. Next, limiting dilution was used for cloning, and cells that consistently showed strong antibody titers were selected as hybridomas for monoclonal antibody production.

(5) Preparation of purified monoclonal antibodies

The monoclonal antibody obtained by intraperitoneal injection (4) into 8- to 10-week-old mice or nude mice treated with pristane [0.5 mL of 2,6,10,14-tetramethylpentadecane administered intraperitoneally for 2 weeks] produced hybridomas. Within 10 to 21 days, the hybridomas developed into cancerous ascites.

Ascites fluid was collected from the mice, and after centrifugation to remove solid components, it was salted out with 40-50% ammonium sulfate. The purified sample was then purified using octanoic acid precipitation, DEAE-agarose column, protein A column, or gel filtration column. The IgG or IgM fraction was collected as purified monoclonal antibodies.

In addition, the monoclonal antibody obtained in (4) was used to generate hybridomas, which were then cultured in RPMI 1640 medium with 10% FBS, and the supernatant was removed by centrifugation. The hybridomas were then suspended in SFM medium and cultured for 3 to 7 days.

The obtained cell suspension is centrifuged, and the supernatant is purified using a protein A column or a protein G column to collect the IgG fraction, which can also yield purified monoclonal antibodies. It should be noted that 5% Daigo’s GF21 can also be added to the hybridoma SFM medium.

Antibody subclasses were determined using an enzyme-linked immunosorbent assay (ELISA) with a subclass typing kit. Protein quantity was quantified using the Laurie method or by calculating the absorbance at 280 nm.

(6) Selection of antibodies

The selection of antibodies is as follows, and is determined by measuring the binding of antibodies to MOG-expressing cells using flow cytometry. MOG-expressing cells can be any cells that express MOG on their cell surface, such as animal cells, animal cell lines, and the MOG-forced expression cell line obtained in (1).

After aliquoting MOG-expressing cells into 96-well plates, serum, hybridoma culture supernatant, or purified antibodies are injected as primary antibodies to initiate the reaction. The reacted cells are then thoroughly washed with PBS containing 1–10% bovine serum albumin (BSA) (hereinafter referred to as BSA-PBS), and then labeled with fluorescent reagents as secondary antibodies to initiate the reaction. After thorough washing with BSA-PBS, the fluorescence intensity of the labeled antibodies is measured using flow cytometry to select antibodies that specifically react with MOG-expressing cells.

Alternatively, antibody selection can be made by measuring the binding of monoclonal antibodies to MOG-expressing cells or MOG proteins using ELISA or surface plasmon resonance assays as described below. MOG proteins can be proteins composed of a subset of MOG domains or proteins tagged with GST or similar tags.

In ELISA, MOG-expressing cells or MOG protein are aliquoted into 96-well plates or similar plates and blocked with BSA-PBS. Serum, hybridoma culture supernatant, or purified antibodies are then aliquoted as the primary antibody to initiate the reaction. Next, after thorough washing with PBS, fluorescently labeled anti-immunoglobulin antibodies are aliquoted as the secondary antibody to initiate the reaction.

Then, after thorough washing with PBS, chromogenic reagent was added. Finally, the chromogenic reaction was terminated with a reaction stop solution, and the absorbance in each well was measured using a microplate reader to select antibodies that specifically react with MOG-expressing cells or MOG protein.

Surface plasmon resonance (SPR) allows for the immobilization of antibodies onto a suitable sensor chip using known operating procedures, enabling the determination of the affinity of antibodies bound to MOG proteins as analytes.

Based on the obtained antibody affinity, antibodies with the desired affinity for MOG proteins can be selected. Alternatively, MOG proteins can be immobilized on a sensor chip, and the affinity of antibodies binding to MOGs can be measured using antibodies as analytes.

Furthermore, antibodies that competitively bind to MOG against the antibodies of the present invention can be obtained by adding the test antibody to the above-described assay system using flow cytometry or ELISA to allow it to react. That is, by screening for antibodies that inhibit the binding of the antibody of the present invention to MOG when the test antibody is added, antibodies that compete with the antibody of the present invention for binding to the amino acid sequence or stereostructure of MOG can be obtained.

In addition, antibodies that bind to epitopes of the antibodies containing the present invention can be obtained by the following method: identifying the epitopes of the antibodies obtained by the above screening method using known methods, preparing synthetic peptides containing the identified epitopes or synthetic peptides with stereoscopic structures mimicking the epitopes, and performing immunization.

Furthermore, antibodies that bind to the same epitope as the antibody of the present invention can be obtained by the following method: identifying the epitope of the antibody obtained by the screening method described above, preparing a synthetic peptide containing a portion of the identified epitope or a synthetic peptide with a stereostructure mimicking the epitope, and then performing immunization.

(7) Obtaining antibodies using phage display

(7-1) Method for preparing antibody phage libraries

In this invention, the antibody phage library can be an immune library, a natural library, or a synthetic library. The methods for preparing each library are described below.

For immune libraries, lymphocytes are collected from animals or patients that have been immunized using the same method as described in (1) above. For natural libraries, lymphocytes are collected from normal animals or healthy individuals, RNA is extracted, and cDNA is synthesized through reverse transcription.

Using this cDNA as a template, PCR was performed for amplification. The amplified antibody gene fragment was then inserted into a phage vector, which was used to transform *E. coli*. When helper phages infected the resulting transformants, an antibody phage library with the antibody gene library was obtained.

In addition, for synthetic libraries, the CDR of the V gene in the genomic DNA or the reconstructed functional V gene is replaced with an oligonucleotide encoding a random amino acid sequence of appropriate length, and E. coli is transformed using a phage vector containing the inserted V gene. When helper phages infect the resulting transformants, an antibody phage library can be obtained.

cDNA and antibody phage libraries derived from lymphocytes can also be commercially available.

Phage vectors that can be used include pCANTAB 5E (Amersham Pharmacia), pUC118/pUC119 vector (TaKaRa), pBlueScript II phage vector (Agilent Technologies), and pKSTV-02 (Miyazaki et al., J. Biochem. 2015; 1).

Helper phages can include M13KO7 helper phage (Invitrogen), VCSM13 interference-resistant helper phage (Agilent Technologies), and R408 interference-resistant helper phage (Agilent Technologies).

Phage display can also be performed using phage vectors. There are peptide phage libraries (such as those produced by New England Biolabs) that use g3p of fibrous phages as the display molecule, and methods that use g7p, g8p, and g9p as the display molecule.

Alternatively, phage display using T7 phage can be employed. Display systems for T7 phage include T7Select vector (Novagen).

(7-2) Selection of antibody phage clones

Selecting antibody phage clones from the antibody phage library prepared in (7-1) can be done using the ELISA method shown below.

MOG was immobilized on an immunotube and blocked with blocking buffer. The antibody-phage library prepared in step (7-1) was added to each well of the tube, and the reaction was initiated. The wells were then washed, fluorescently labeled anti-phage antibodies were added, and the reaction was initiated again. The wells were washed once more, and chromogenic solution was added. The chromogenic reaction was then terminated with stop solution, and the absorbance of each well was measured using a microplate reader. This process allowed for the selection of antibody-phage clones that bind to MOG.

2. Production of recombinant antibodies

As examples of recombinant antibody production, the following methods illustrate the preparation of human chimeric antibodies and humanized antibodies. Recombinant mouse antibodies, rat antibodies, rabbit antibodies, hamster antibodies, camel antibodies, llama antibodies, alpaca antibodies, human antibodies, various chimeric antibodies, and heavy chain antibodies can also be prepared using the same methods.

(1) Construction of vectors for expressing recombinant antibodies

Recombinant antibody expression vectors are animal cell expression vectors that integrate DNA encoding CH and CL of human antibodies. They can be constructed by cloning the DNA encoding CH and CL of human antibodies into the animal cell expression vectors, respectively.

The C region of a human antibody can use the CH and CL of any human antibody. For example, the CH of the γ1 subclass and the CL of the κ class can be used. The DNA encoding the CH and CL of the human antibody is cDNA, but chromosomal DNA consisting of exons and introns can also be used.

For animal cell expression vectors, any expression vector can be used as long as it can integrate and express the gene encoding the C region of the human antibody. For example, pAGE107 [Cytotechnol., 3, 133 (1990)], pAGE103 [J. Biochem., 101, 1307 (1987)], pHSG274 [Gene, 27, 223 (1984)], pKCR [Proc. Natl. Acad. Sci. USA, 78, 1527 (1981)], pSG1bd2-4 [Cytotechnol., 4, 173 (1990)], or pSE1UK1Sed1-3 [Cytotechnol., 13, 79 (1993)] can be used.

Promoters and enhancers in animal cell expression vectors include the early promoter of SV40 [J. Biochem., 101, 1307 (1987)], the promoter of Moloney mouse leukemia virus LTR [Biochem. Biophys. Res. Commun., 149, 960 (1987)], or the promoter and enhancer of immunoglobulin H chain [Cell, 41, 479 (1985)], etc.

Based on considerations such as the ease of constructing recombinant antibody expression vectors, the ease of introduction into animal cells, and achieving a balance in the expression levels of antibody H and L chains within animal cells, recombinant antibody expression vectors using this type (tandem type) have antibody H and L chains on the same vector [J. Immunol. Methods, 167, 271 (1994)]. However, it is also possible to use vectors with antibody H and L chains on separate vectors. Tandem recombinant antibody expression vectors include pKANTEX93 (International Publication No. 97/10354) and pEE18 [Hybridoma, 17, 559 (1998)].

(2) Obtaining cDNA encoding the V region of antibodies derived from animals other than humans and analyzing its amino acid sequence.

The cDNA encoding non-human antibodies VH and VL can be obtained and their amino acid sequences analyzed as follows.

(2-1) Cases of obtaining antibodies using the hybridoma method

mRNA was extracted from hybridoma cells that produce non-human antibodies, and cDNA was synthesized. The synthesized cDNA was cloned into vectors such as bacteriophages or plasmids to create a cDNA library.

Using DNA encoding the C or V region of a non-human antibody as a probe, recombinant phages or recombinant plasmids encoding VH or VL cDNA were isolated from the aforementioned library. The complete base sequence of the target non-human antibody VH or VL on the recombinant phage or recombinant plasmid was determined, and the complete amino acid sequence of VH or VL was deduced from the base sequence.

Animals other than humans that produce hybridoma cells that generate non-human antibodies can be used, such as mice, rats, hamsters, rabbits, llamas, camels, or alpacas, but any animal can be used as long as hybridoma cells can be produced.

When preparing total RNA from hybridoma cells, kits such as the guanidine isothiocyanate-cesium trifluoroacetate method [Methods in Enzymol., 154, 3 (1987)] or the RNA easy kit (manufactured by Qiagen) can be used.

When preparing mRNA from total RNA, kits such as the oligomeric (dT) immobilized cellulose column method [Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press (1989)] or the Oligo-dT30<Super> (registered trademark) mRNA purification kit (manufactured by Takara Bio Inc.) can be used. Alternatively, kits such as the Fast Track (registered trademark) mRNA isolation kit (manufactured by Invitrogen) or the QuickPrep (registered trademark) mRNA purification kit (manufactured by Pharmacia) can be used to prepare mRNA from hybridoma cells.

cDNA synthesis and cDNA library preparation can be performed using well-known methods [Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press (1989); Current Protocols in Molecular Biology, Appendix 1, John Willie & Son Press (1987-1997)]; or kits such as the SuperScript plasmid system (manufactured by Invitrogen) or the ZAP-cDNA (registered trademark) synthesis kit (manufactured by Stratagene) for cDNA synthesis and plasmid cloning.

When constructing a cDNA library, any vector can be used to integrate cDNA synthesized using mRNA extracted from hybridoma cells as a template, as long as it can integrate the cDNA. For example, ZAP Express [Strategies, 5, 58 (1992)], pBluescript II SK(+) [Nucleic Acids Research, 17, 9494 (1989)], λZAPII (manufactured by Stratagene), λgt10, λgt11 [DNA Cloning: A Practical Approach, I, 49 (1985)], Lambda BlueMid (manufactured by Clontech), λExCell, pT7T3-18U (manufactured by Pharmacia), pCD2 [Mol.Cell.Biol., 3, 280 (1983)] or pUC18 [Gene, 33, 103 (1985)], etc.

E. coli that can import cDNA libraries constructed using phage or plasmid vectors can be used, as long as the cDNA library can be imported, expressed, and maintained. For example, XL1-Blue MRF’ [Strategies, 5, 81 (1992)], C600 [Genetics, 39, 440 (1954)], Y1088, Y1090 [Science, 222, 778 (1983)], NM522 [J.Mol.Biol., 166, 1 (1983)], K802 [J.Mol.Biol., 16, 118 (1966)], or JM105 [Gene, 38, 275 (1985)] can be used.

When selecting cDNA clones encoding non-human antibodies VH or VL from a cDNA library, colony hybridization or plaque hybridization using isotope- or fluorescently labeled probes can be used [Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press (1989)].

Alternatively, primers can be prepared to use cDNA synthesized from mRNA or a cDNA library as a template for polymerase chain reaction (PCR) [Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press (1989); Current Protocols in Molecular Biology, Appendix 1, John Willie & Son Press (1987-1997)], thereby preparing cDNA encoding VH or VL.

The selected cDNA is cut with an appropriate restriction enzyme and cloned into a plasmid such as pBluescript SK(-) (manufactured by Stratagene). The base sequence of the cDNA is then determined using commonly used base sequence analysis methods. For example, after performing a dideoxy method [Proc. Natl. Acad. Sci. USA, 74, 5463 (1977)], an automated base sequence analysis device such as an ABI PRISM3700 (manufactured by PE Biosystems) or an A.L.F. DNA sequencer (manufactured by Pharmacia) can be used.

(2-2) Cases of obtaining antibodies using phage display method

Using DNA from the encoding vector portion or the V region as probes, the complete base sequence of VH or VL is determined from the plasmid vector of the selected phage clone, and the complete amino acid sequence of VH or VL is deduced from the base sequence.

In either the hybridoma method or the phage display method, the complete amino acid sequences of VH and VL are deduced from the determined base sequences and compared with the complete amino acid sequences of VH and VL of known antibodies [Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services (1991)] to confirm whether the obtained cDNA encodes the complete amino acid sequences of VH and VL of the antibody containing the secretion signal sequence.

The length of the secretion signal sequence and the N-terminal amino acid sequence can be estimated by comparing the complete amino acid sequences of the VH and VL of antibodies containing the secretion signal sequence with those of known antibodies [Sequences of Proteins of Immunological Interest, USDept. Health and Human Services (1991)]. This allows us to determine their subclass.

In addition, the amino acid sequences of each CDR for VH and VL can also be identified by comparing them with the amino acid sequences of VH and VL of known antibodies [Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services (1991)].

In addition, using the obtained complete amino acid sequences of VH and VL, homology searches can be performed on any database such as SWISS-PROT or PIR-Protein using methods such as BLAST [J.Mol.Biol.,215,403(1990)] to confirm whether the complete amino acid sequences of VH and VL are new sequences.

(3) Construction of human chimeric antibody expression vector

The cDNA encoding VH or VL of non-human antibodies is cloned into the upstream of each gene encoding CH or CL of human antibodies in the gene recombinant antibody expression vector obtained in (1), thereby constructing a human chimeric antibody expression vector.

In order to link the 3' end of the cDNA encoding a non-human antibody VH or VL to the 5' end of a human antibody CH or CL, VH and VL cDNAs are designed such that the base sequence of the linker encodes an appropriate amino acid and becomes an appropriate restriction enzyme recognition sequence.

The cDNAs of the prepared VH and VL were cloned upstream of the genes encoding the human antibody CH or CL of the gene recombinant antibody expression vector obtained in (1) so that they were expressed in an appropriate form, thereby constructing a human chimeric antibody expression vector.

Alternatively, synthetic DNA with appropriate restriction enzyme recognition sequences at both ends can be used to amplify cDNA encoding non-human antibodies VH or VL using PCR, and then clone them into the gene recombinant antibody expression vector obtained in (1).

(4) Construction of cDNA encoding the V region of humanized antibody

The cDNA encoding the humanized antibody VH or VL can be constructed as follows.

Select the amino acid sequence of the FR of the human antibody VH or VL, respectively, which is the CDR sequence of the non-human antibody VH or VL for transplantation. Any amino acid sequence can be used as long as it is derived from the FR sequence of the human antibody.

For example, the amino acid sequences of the FR of human antibodies registered in databases such as the Protein Data Bank, or the common amino acid sequences of various subclasses of the FR of human antibodies [Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services (1991)]. In order to suppress the decrease in antibody binding activity, the amino acid sequence of the FR with the highest possible homology (at least 60%) to the FR sequence of the original antibody's VH or VL is selected.

Next, the amino acid sequence of the original antibody's CDR was transplanted into the FR amino acid sequence of the selected human antibody's VH or VL, respectively, to design the amino acid sequences of the humanized antibody's VH or VL. Considering the frequency of codon usage in the antibody gene's base sequence [Sequences of Proteins of Immunological Interest, USDept. Health and Human Services (1991)], the designed amino acid sequences were converted into DNA sequences, and DNA sequences encoding the amino acid sequences of the humanized antibody's VH or VL were designed.

Based on the designed DNA sequence, multiple synthetic DNA sequences, each approximately 100 bases in length, are synthesized, and these synthetic DNA sequences are used for PCR reactions. In this case, considering the reaction efficiency in the PCR reaction and the length of DNA that can be synthesized, it is preferable to design 6 synthetic DNA sequences for each of VH and VL.

Furthermore, by introducing appropriate restriction enzyme recognition sequences into the 5' or 3' ends of the synthetic DNA located at both ends, it is possible to easily clone the cDNA encoding the humanized antibody VH or VL into the gene recombinant antibody expression vector obtained in (1).

After the PCR reaction, the amplification products were cloned into plasmids such as pBluescript SK(-) (manufactured by Stratagene), and the base sequence was determined by the same method as described in (2), thereby obtaining a plasmid with a DNA sequence having the desired amino acid sequence encoding the VH or VL of the humanized antibody.

Alternatively, DNA obtained by synthesizing full-length VH and full-length VL in the form of one long chain DNA each based on the designed DNA sequence can be used instead of the PCR amplification products described above. Furthermore, by introducing appropriate restriction enzyme recognition sequences at both ends of the synthesized long chain DNA, the cDNA encoding the humanized antibody VH or VL can be easily cloned into the gene recombinant antibody expression vector obtained in (1).

(5) Changes in the amino acid sequence of the V region of humanized antibodies

For humanized antibodies, when only the CDRs of the VH and VL of a non-human antibody are transplanted into the FRs of the VH and VL of a human antibody, their antigen-binding activity is reduced compared to the original non-human antibody [BIO/TECHNOLOGY, 9, 266 (1991)].

In humanized antibodies, amino acid residues in the VH and VL FR amino acid sequences of human antibodies that are directly related to antigen binding, amino acid residues that interact with CDR amino acid residues, and amino acid residues that are indirectly related to antigen binding while maintaining the antibody's stereostructure are identified. These amino acid residues are then replaced with the original non-human antibody amino acid residues, thereby increasing the reduced antigen binding activity.

To identify the amino acid residues of the antigen-binding receptor (FR) associated with antigen-binding activity, the stereostructure of the antibody can be constructed and analyzed using methods such as X-ray crystallography [J. Mol. Biol., 112, 535 (1977)] or computer modeling [Protein Engineering, 7, 1501 (1994)]. Furthermore, various attempts are made to create multiple alterations of each antibody and repeatedly study their correlation with their respective antigen-binding activities. In this way, humanized antibodies with the desired antigen-binding activity can be obtained.

The amino acid residues of VH and VL of human antibodies can be altered by performing a PCR reaction using synthetic DNA as described in (4). The amplification products after the PCR reaction are then used to determine the base sequence by the method described in (2), thereby confirming that the target alteration has been performed.

(6) Construction of humanized antibody expression vector

By cloning the cDNA of the constructed recombinant antibody VH or VL into the upstream of each gene encoding the human antibody CH or CL of the recombinant antibody expression vector obtained in (1), a humanized antibody expression vector can be constructed.

For example, by introducing appropriate restriction enzyme recognition sequences into the 5' or 3' ends of the synthetic DNA used to construct the humanized antibodies VH or VL obtained in (4) and (5), the sequences are cloned upstream of the genes encoding the human antibodies CH or CL in the gene recombinant antibody expression vector obtained in (1), so that they are expressed in an appropriate form.

(7) Transient expression of recombinant antibodies

Using the recombinant antibody expression vectors obtained in (3) and (6) or expression vectors obtained by modifying them, the transient expression of recombinant antibodies can be carried out, which can efficiently evaluate the antigen-binding activity of various human chimeric antibodies and humanized antibodies produced.

The host cell into which the expression vector is introduced can be any cell that can express the recombinant antibody, such as COS-7 cells [American Center for Type Culture Collection (ATCC) No.: CRL1651][Methods in Nucleic Acids Res., CRC Press, 283 (1991)].

When introducing expression vectors into COS-7 cells, the DEAE-dextran method [Methods in Nucleic Acids Res., CRC Press (1991)] or liposome transfection method [Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)] can be used.

After the expression vector was introduced, the expression level and antigen-binding activity of the recombinant antibody in the culture supernatant were determined using enzyme immunoassay [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Antibodies - A Laboratory Manual, Cold Spring Harbor Laboratory (1988); Monoclonal Antibody Laboratory Manual, Kodansha Scientific (1987)].

(8) Obtaining transformants that stably express recombinant antibodies and preparing recombinant antibodies

By introducing the recombinant antibody expression vector obtained in (3) and (6) into appropriate host cells, a transformant strain that stably expresses the recombinant antibody can be obtained.

The introduction of expression vectors into host cells is achieved using electroporation [Japanese Patent Application Publication No. 2-257891; Cytotechnology, 3, 133 (1990)], etc.

Any host cell that can express recombinant antibody expression vectors can be used. For example, CHO-K1 (ATCC CCL-61), DUKXB11 (ATCC CCL-9096), Pro-5 (ATCC CCL-1781), CHO-S (Life Technologies, Cat#11619), rat myeloma cells YB2/3HL.P2.G11.16Ag.20 (ATCC number: CRL1662, or also known as YB2/0), mouse myeloma cells NS0, mouse myeloma cells SP2/0-Ag14 (ATCC number: CRL1581), mouse P3X63-Ag8.653 cells (ATCC number: CRL1580), and CHO cells (CHO/DG44 cells) deficient in the dihydrofolate reductase gene (hereinafter referred to as dhfr) [Proc. Natl. Acad. Sci. USA, 77, 4216 (1980)] were used.

Alternatively, host cells with reduced or absent activity, such as enzymes or proteins related to the synthesis of intracellular glyconucleotide GDP-fucose, enzymes or proteins related to the modification of the glycan chain by α-bonding of fucose at the 6-position of the N-acetylglucosamine at the reducing end of the N-glycosidic complex glycan chain, or proteins related to the transport of intracellular glyconucleotide GDP-fucose to the Golgi apparatus, can be used. Examples include CHO cells with α1,6-fucosyltransferase gene deficiency (International Publication No. 2005/035586, International Publication No. 02/31140), and Lec13 cells that have acquired lectin resistance [Somatic Celland Molecular Genetics, 12, 55 (1986)].

After the expression vector was introduced, the transformant strains that stably expressed the recombinant antibody were cultured in an animal cell culture medium containing agents such as G418 sulfate (hereinafter referred to as G418) and selected accordingly (Japanese Patent Application Publication No. 2-257891).

Animal cell culture media include RPMI 1640 medium (manufactured by Invitrogen), GIT medium (manufactured by Nippon Pharmaceutical Co., Ltd.), EX-CELL 301 medium (manufactured by JRH), IMDM medium (manufactured by Invitrogen), or hybridoma-SFM medium (manufactured by Invitrogen), or mediums containing various additives such as FBS.

The resulting transformants are cultured in a culture medium to allow recombinant antibodies to be expressed and accumulated in the culture supernatant. The expression level and antigen-binding activity of the recombinant antibodies in the culture supernatant can be measured using methods such as ELISA. Alternatively, the expression level of recombinant antibodies produced by the transformants can be increased using a dhfr gene amplification system (Japanese Patent Application Laid-Open No. 2-257891).

Recombinant antibodies can be purified from the culture supernatant of transformants using a protein A column [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Antibodies - A Laboratory Manual, Cold Spring Harbor Laboratory (1988)]. Alternatively, purification can be achieved by combining methods used in protein purification such as gel filtration, ion exchange chromatography, and ultrafiltration.

The molecular weight of the H chain, L chain, or the entire antibody molecule of the purified recombinant antibody can be determined using polyacrylamide gel electrophoresis [Nature, 227, 680 (1970)] or Western blotting [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Antibodies - A Laboratory Manual, Cold Spring Harbor Laboratory (1988)].

(9) Methods for preparing antibody fragments

The antibody fragments of the present invention can be prepared according to known methods. The antibody fragments of the present invention can be prepared by cutting the antibody prepared according to the methods described in (1) to (8) above using enzymes or the like, or by preparing the base sequence encoding the desired antibody fragment and preparing it using genetic engineering methods.

(10) Methods for preparing monovalent antibodies

In this invention, the monovalent antibody can be prepared using the methods described in International Publication No. 2014/054804, International Publication No. 2011/090754, International Publication No. 2007/048037 and International Publication No. 2012/116927, etc.

(11) Methods for manufacturing bispecific or multispecific antibodies

The bispecific or multispecific antibodies of the present invention can be manufactured according to the antibody manufacturing method described above. For example, International Publication No. 2009/131239, International Publication No. 2014/054804, International Publication No. 01/077342, U.S. Patent Application Publication No. 2007/0071675, International Publication No. 2007/024715, Wu et al., [Nature Biotechnology, 2007, 25(11), pp. 1290-1297], Labrijn et al., [PNAS 2013, vol. 110, no. 13, pp. 5145-5150], and Jong et al., [http://dx.doi.org/10.137]. The method described in [1/journal.pbio.1002344], Kontermann et al., [mAbs 2012, vol.4, issue2, p182-197], Spiess et al., [Molecular Immunology 67(2015)95-106], Ridgway et al., [Protein Engineering, 1996 vol.9 no.7 pp617-621], International Publication No. 2009/080251, International Publication No. 2010/151792 and International Publication No. 2014/033074, etc., is used to prepare it.

For example, an expression vector for a bispecific antibody formed by fusing an MOG-binding scFv to the C-terminus of an IgG antibody that binds to antigens present in the brain can be prepared using the method described below. This bispecific antibody can be prepared according to the antibody expression and purification methods described above. Furthermore, other bispecific antibodies formed by fusing antibody fragments to the C-terminus of an antibody can also be prepared using the same method.

Using the gene for synthesizing the heavy chain constant region of an IgG antibody that binds to antigens present in the brain as a template, a gene fragment of the CH1-hinge-CH2-CH3-linker region was amplified by PCR. Next, using the base sequence of an antibody that binds to MOG as a template, the base sequence of the scFv region, formed by combining the VH and VL regions of the antibody with appropriate linkers, was prepared using PCR or similar methods. The two regions were then bound together using PCR or similar methods, and the resulting gene fragment was inserted into a suitable vector such as a pCI vector.

In addition, gene fragments of the light chain region (VL and CL) of IgG antibodies that bind to antigens present in the brain and gene fragments of the VH of the antibody were amplified by PCR using appropriate templates and inserted into appropriate positions in the aforementioned vector.

In addition, the bispecific antibody of the present invention can also be prepared by using a chemical method to bind an antigen-binding site containing an antibody fragment to an IgG antibody.

3. Evaluation of the activity of the antibody or the antibody fragment.

In this invention, the activity evaluation of the antibody or the antibody fragment can be performed as follows.

(1) Binding activity to MOG

The binding activity of the antibody or antibody fragment of the present invention to MOG can be determined using flow cytometry, ELISA, and surface plasmon resonance detection as described in 1-(6) above. Alternatively, it can be determined using a fluorescent antibody method [Cancer Immunol. Immunother., 36, 373 (1993)].

When the antibody or antibody fragment of the present invention is a monovalent antibody that binds to MOG, the binding activity of the monovalent antibody to MOG can be determined using the same method. When the antibody or antibody fragment of the present invention is a bispecific or multispecific antibody that binds to both MOG and antigens present in the brain, the binding activity of the bispecific or multispecific antibody to MOG or antigens present in the brain can be determined using the same method.

(2) Methods for measuring brain retention

The brain retention of the antibody or antibody fragment of the present invention can be determined using the methods described below.

Examples include: administering an antibody or antibody fragment to an animal, recovering brain tissue several days later, homogenizing it, determining the concentration of the antibody or antibody fragment in the supernatant after centrifugation, and calculating the amount of antibody or antibody fragment per unit weight of brain tissue; or using the recovered brain tissue to detect the presence of the antibody or antibody fragment using known immunological methods; and so on. Additionally, examples include: administering a pharmacologically acceptable labeled antibody or antibody fragment to an animal and using an in vivo imaging system to detect the presence of the antibody or antibody fragment over time.

The animals used may be selected from suitable animals appropriate to the use of the antibody or antibody fragment of the present invention.

(3) Measurement methods for ADCC and CDC

The antibody or antibody fragment of the present invention can be used to measure CDC or ADCC in human MOG-expressing cells or cells expressing MOG and antigens present in the brain using known assay methods [Cancer Immunol. Immunother., 36, 373 (1993); Current protocols in Immunology, Chapter 7. Immunologic studies in humans, Editor, John E, Coligan et al., John Willie & Son Publishing Co. (1993)].

4. Methods for controlling the effector activity of antibodies or antibody fragments

Methods for controlling the effector activity of the antibody or antibody fragment of the present invention include: methods for controlling the amount of fucose (also known as core fucose) present at the reducing end of an N-acetylglucosamine (GlcNAc) bonded to asparagine (Asn) at position 297 of the Fc region of the antibody or the antibody fragment containing Fc, which are bound by an α1,6 bond (International Publication Nos. 2005/035586, 2002/31140, and 00/61739); or methods for controlling the activity by altering the amino acid residues in the Fc region of the antibody or the antibody fragment. Using any of these methods with the antibody or antibody fragment of the present invention allows for control of effector activity.

Effector activity refers to antibody-dependent activity caused by an antibody or the Fc region of an antibody fragment. Known examples include ADCC activity, CDC activity, or antibody-dependent phagocytosis (ADP activity) produced by phagocytes such as macrophages or dendritic cells.

As a method for measuring effector activity, for example, target cells, human peripheral blood mononuclear cells (PBMCs) as effector cells, and a target cell-specific antibody or fragment thereof can be mixed, incubated for about 4 hours, and then the free lactate dehydrogenase (LDH), an indicator of cell damage, can be measured. Alternatively, effector activity can be measured using the free ^51Cr method or flow cytometry.

By controlling the content of fucose, the core of the N-binding complex glycan of the antibody's Fc, the effector activity of the antibody or the antibody fragment containing the Fc can be increased or decreased. As a method to reduce the content of fucose bound to the N-binding complex glycan bound to the Fc of the antibody or the antibody fragment, an antibody or antibody fragment without fucose binding can be obtained by expressing the antibody or the antibody fragment using CHO cells deficient in the α1,6-fucosyltransferase gene. The antibody or antibody fragment without fucose binding exhibits high ADCC activity.

On the other hand, as a method to increase the content of fucose that binds to the N-binding complex glycans bound to the Fc of an antibody or antibody fragment, an antibody or antibody fragment bound to fucose can be obtained by expressing the antibody or antibody fragment using a host cell infused with the α1,6-fucotransferase gene. The antibody or antibody fragment bound to fucose exhibits lower ADCC activity than the antibody or antibody fragment not bound to fucose.

Additionally, ADCC or CDC activity can be increased or decreased by altering the amino acid residues in the Fc region of the antibody or antibody fragment. For example, the CDC activity of the antibody or antibody fragment can be increased by using the amino acid sequence of the Fc region described in U.S. Patent Application Publication No. 2007/0148165.

In addition, by making the amino acid changes described in U.S. Patent No. 6,737,056, U.S. Patent No. 7,297,775, or U.S. Patent No. 7,317,091, ADCC activity or CDC activity can be increased or decreased.

Furthermore, the antibody or antibody fragment of the present invention also includes an antibody or antibody fragment whose responsiveness to the Fc receptor is controlled by altering the amino acid or glycan in the constant region contained in the antibody or antibody fragment as described in, for example, Japanese Patent Application Publication No. 2013-165716 or Japanese Patent Application Publication No. 2012-021004, thereby controlling the half-life of the antibody or antibody fragment in the blood.

Furthermore, by combining the above methods for use on an antibody or antibody fragment, it is possible to obtain an antibody or antibody fragment that controls effector activity and blood half-life.

5. Treatment methods for diseases using the antibodies or antibody fragments of the present invention.

The antibody or antibody fragment of the present invention can be used to treat brain diseases in animals that express MOG in the brain.

Examples of brain diseases include Alzheimer's disease, Alzheimer's disease in its early stages, Huntington's disease, Parkinson's disease, brain tumors, multiple sclerosis, muscular dystrophy, amyotrophic lateral sclerosis, multiple system atrophy, progressive supranuclear palsy, nigrostriatal degeneration, olivopontocerebellar atrophy, spinobulbar muscular atrophy, spinocerebellar degeneration, cerebrovascular diseases, epilepsy, migraine, ADHD, Creutzfeldt-Jakob disease, corticobasal degeneration, lysosomal storage diseases, depression, and dystonia.

The brain diseases that the antibodies or antibody fragments of the present invention can treat vary depending on the antigens bound to the antibodies or antibody fragments of the present invention, and the types of molecules used to modify the antibodies or antibody fragments using the fusion antibodies or fusion antibody fragments of the present invention.

Therapeutic agents containing the antibodies or antibody fragments of the present invention may contain only the antibody or antibody fragment as an active ingredient, but are generally preferably provided as pharmaceutical preparations mixed with one or more pharmacologically acceptable carriers and manufactured using methods known in the field of pharmaceutical formulation.

Examples of routes of administration include oral administration, intraoral administration (oral, respiratory, rectal, subcutaneous, intramuscular, intravenous, intraperitoneal, intradermal, nasal, intramedullary, or intravenous administration). Examples of forms of administration include sprays, capsules, tablets, powders, granules, syrups, emulsions, suppositories, injections, ointments, and patches.

Examples of preparations suitable for oral administration include emulsions, syrups, capsules, tablets, powders, and granules.

Liquid preparations such as emulsions or syrups are manufactured using sugars such as water, sucrose, sorbitol or fructose, glycols such as polyethylene glycol or polypropylene glycol, oils such as sesame oil, olive oil or soybean oil, preservatives such as parabens, or flavorings such as strawberry flavoring or mint flavoring as additives.

Capsules, tablets, powders, or granules are manufactured using excipients such as lactose, glucose, sucrose, or mannitol; disintegrants such as starch or sodium alginate; lubricants such as magnesium stearate or talc; binders such as polyvinyl alcohol, hydroxypropyl cellulose, or gelatin; surfactants such as fatty acid esters; or plasticizers such as glycerin.

As preparations suitable for non-oral administration, these include injections, suppositories, and sprays. Injections are manufactured using carriers such as saline solutions, glucose solutions, or mixtures of both. Suppositories are manufactured using carriers such as cocoa butter, hydrogenated fats, or carboxylic acids.

The spray is manufactured using a carrier that does not irritate the recipient's oral and respiratory mucosa and disperses the antibody or antibody fragment of the present invention in the form of fine particles for easy absorption. Examples of carriers include lactose or glycerol. Alternatively, it can be manufactured in aerosol or dry powder form. Furthermore, the aforementioned non-oral formulations may also include the ingredients exemplified as additives in formulations suitable for oral administration.

6. A method for detecting or measuring antigens present in the brain using the antibody or antibody fragment of the present invention, or a method for diagnosing a disease.

By using the antibody or antibody fragment of the present invention, it is possible to detect or measure MOG, or MOG and antigens present in the brain. Furthermore, by detecting or measuring MOG, or MOG and antigens present in the brain, it is possible to diagnose brain diseases in animals expressing MOG in the brain.

Examples of brain diseases include Alzheimer's disease, Alzheimer's disease in its early stages, Huntington's disease, Parkinson's disease, brain tumors, multiple sclerosis, muscular dystrophy, amyotrophic lateral sclerosis, multiple system atrophy, progressive supranuclear palsy, nigrostriatal degeneration, oligopontocerebellar atrophy, spinobulbar muscular atrophy, spinocerebellar degeneration, cerebrovascular diseases, epilepsy, migraine, ADHD, Creutzfeldt-Jakob disease, corticobasal degeneration, lysosomal storage disease, depression, and dystonia. The brain diseases that the antibodies or antibody fragments of the present invention can diagnose vary depending on the antigens bound to the antibodies or antibody fragments of the present invention, and the types of molecules used to modify the antibodies or antibody fragments using the fusion antibodies or fusion antibody fragments of the present invention.

The diagnosis of brain diseases in animals expressing MOGs in the brain can be made, for example, by detecting or measuring MOGs present in the brain of the patient or diseased animal using immunological methods. Alternatively, diagnosis can be made by detecting MOGs expressed or present on cells in the brain of the patient or diseased animal using immunological methods such as flow cytometry.

When using a monovalent antibody that binds to MOG as the antibody or antibody fragment of the present invention, MOG in the brain can be measured using the same method as described above. When using a bispecific or multispecific antibody that binds to both MOG and antigens present in the brain as the antibody or antibody fragment of the present invention, MOG in the brain or antigens present in the brain can be detected or measured using the same method as described above.

Immunological methods refer to methods that use labeled antigens or antibodies to detect or measure the amount of antibody or antigen. Examples include radiolabeled antibody methods, enzyme-linked immunosorbent assays (ELISA), fluorescence immunoassays, luminescent immunoassays, Western blotting, and physicochemical methods.

The radiolabeled immunosorbent assay (RIA) involves reacting the antibody or antibody fragment of the present invention with an antigen or cells expressing the antigen, further reacting with a radiolabeled anti-immunoglobulin antibody or antibody fragment, and then measuring using a scintillation counter or the like.

Enzyme-linked immunosorbent assay (ELISA) involves reacting the antibody or antibody fragment of the present invention with an antigen or cells expressing the antigen, further reacting with an anti-immunoglobulin antibody or antibody fragment labeled with an enzyme, then adding a substrate and measuring the absorbance of the reaction solution using a spectrophotometer. For example, a sandwich ELISA method can be used. As a label used in ELISA, enzyme labels known to the public [ELISA, Medical School (1987)] can be used.

Examples of labeling methods include alkaline phosphatase, peroxidase, luciferase, or biotin. Sandwich ELISA involves binding an antibody to a solid phase, which then captures the antigen to be detected or measured, and a second antibody reacts with the captured antigen.

In the ELISA method described above, two antibodies with different antigen recognition sites are prepared to recognize the antigen to be detected or measured. The first antibody is pre-adsorbed onto a plate (e.g., a 96-well plate), and the second antibody is pre-labeled with fluorescent substances such as FITC, enzymes such as peroxidase, or biotin.

Cells or their lysate, tissues or their lysate, cell culture supernatant, serum, pleural fluid, ascites, or eye fluid isolated from a living organism are reacted on the plate adsorbed with the first antibody. Then, the second antibody is reacted, followed by a detection reaction corresponding to the labeled substance. The antigen concentration in the test sample is calculated using a standard curve prepared by serially diluting the antigen to a known concentration.

The antibodies used in sandwich ELISA can be either polyclonal or monoclonal antibodies. Alternatively, antibody fragments such as Fab, Fab', or F(ab) ₂ can be used instead of antibodies. The combination of two antibodies used in sandwich ELISA can be a combination of monoclonal antibodies or antibody fragments that recognize different epitopes, or a combination of polyclonal antibodies and monoclonal antibodies or antibody fragments of these antibodies.

Fluorescent immunoassays utilize methods described in references such as [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Monoclonal Antibody Laboratory Manual, Kodansha Scientific (1987)]. As the markers used in fluorescent immunoassays, known fluorescent labels [Fluorescent Antibody Method, SoftScience (1983)] can be used. For example, FITC or RITC can be used.

The luminescent immunoassay is performed using the methods described in the literature [Bioluminescence and Chemiluminescence Clinical Examination 42, Hirokawa Shoten (1998)]. Commonly known luminescent markers used in the luminescent immunoassay include acridinium esters or cyclophosphamide.

In Western blotting, antigens or cells expressing antigens are graded using SDS (sodium dodecyl sulfate)-PAGE (polyacrylamide gel) [Antibodies-A Laboratory Manual, Cold Spring Harbor Laboratory (1988)]. The gel is then imprinted onto a polyvinylidene fluoride (PVDF) or nitrocellulose membrane. Antibodies or antibody fragments that recognize the antigen react with the membrane, thereby reacting with anti-mouse IgG antibodies or binding fragments labeled with fluorescent substances such as FITC, enzymes such as peroxidase, or biotin. The labeling is then made visible, and the results are measured. An example is shown below.

Cells or tissues expressing peptides with the MOG amino acid sequence were lysed, and the protein amount in each lane was reduced to 0.1–30 μg under reducing conditions. SDS-PAGE electrophoresis was then performed. The electrophoretically extracted proteins were transferred to a PVDF membrane and incubated in PBS containing 1–10% BSA (hereinafter referred to as BSA-PBS) at room temperature for 30 minutes to perform a blocking operation.

Here, the antibody or antibody fragment of the present invention is reacted with PBS containing 0.05 to 0.1% Tween-20 (hereinafter referred to as Tween-PBS), and peroxidase-labeled goat anti-mouse IgG is reacted at room temperature for 2 hours.

The strips containing the antibody or antibody fragment of the present invention were washed with Tween-PBS and detected using ECL protein immunoblotting assay reagent (manufactured by Amasia Corporation), thereby detecting peptides with the amino acid sequence of MOG.

As an antibody or antibody fragment used in detection using Western blotting, an antibody or antibody fragment capable of binding to a polypeptide that does not retain its native three-dimensional structure is used.

Physicochemical methods, for example, involve forming aggregates by binding the MOG, which serves as an antigen, to the antibody or antibody fragment of the present invention and detecting the aggregates. Other physicochemical methods include capillary methods, one-way immunodiffusion, immunoturbidimetry, or latex immunoturbidimetry [Summary of Clinical Examination Methods, Kinhara Publishing (1998)].

In latex immunoturbidimetry, a carrier such as polystyrene latex with a particle size of approximately 0.1 μm to approximately 1 μm, sensitized with antibodies or antigens, is used. When the corresponding antigen or antibody induces an antigen-antibody reaction, the scattered light in the reaction solution increases, while the transmitted light decreases. This change is detected in the form of absorbance or integrating sphere turbidity, thereby determining the antigen concentration in the sample.

The detection or assay of cells expressing MOG can be performed using known immunological assays, preferably immunoprecipitation, immunocellular staining, immunotissue staining, or fluorescent antibody staining.

In the immunoprecipitation method, after reacting MOG-expressing cells or the antibody or antibody fragment of the present invention with the present invention, a carrier such as protein G-agarose, which has the specific binding ability to immunoglobulins, is added to precipitate the antigen-antibody complex. Alternatively, the following method can also be used.

The antibody or antibody fragment of the present invention is immobilized on a 96-well ELISA plate and then blocked with BSA-PBS. If the antibody is in an unpurified state, such as hybridoma culture supernatant, anti-mouse immunoglobulin, anti-rat immunoglobulin, protein A, or protein G is pre-immobilized on a 96-well ELISA plate, blocked with BSA-PBS, and then hybridoma culture supernatant is dispensed to allow binding.

Next, after discarding the BSA-PBS and thoroughly washing with PBS, the lysate of cells or tissues expressing human MOG was used for reaction. Immunoprecipitates were extracted from the thoroughly washed plate using SDS-PAGE with sample buffer and detected by the aforementioned Western blotting method.

The immunocellular staining method or immunotissue staining method is as follows: cells or tissues expressing antigens are treated with surfactants or methanol, etc., as appropriate, to make the antibodies permeable. Then, they are reacted with the antibodies of the present invention, and then reacted with anti-immunoglobulin antibodies or their binding fragments that have been labeled with fluorescent markers such as FITC, enzyme markers such as peroxidase, or biotin markers. The markers are then made visible and observed under a microscope.

Alternatively, detection can be performed using fluorescent antibody staining [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Monoclonal Antibody Laboratory Manual, Kodansha Scientific (1987)], where cells are reacted with fluorescently labeled antibodies and analyzed using flow cytometry. In particular, the antibody or antibody fragment of the present invention can be used to detect cells that maintain their native three-dimensional structure using fluorescent antibody staining.

In addition, in fluorescent antibody staining methods, when using systems such as the FMAT8100HTS system (manufactured by Applied Biosystems), the amount of antigen or antibody can be measured without separating the formed antibody-antigen complex from the free antibody or antigen that did not participate in the formation of the antibody-antigen complex.

The present invention will be described in more detail below with reference to embodiments, but the present invention is not limited to the embodiments described below.

Example

[Example 1] Obtaining anti-MOG antibodies

(1) Obtaining antibodies using human antibody phage libraries

The VH and VL gene fragments were amplified from cDNA derived from human PBMCs using PCR. The VH and VL gene fragments were then inserted into the phage vector pCANTAB 5E (manufactured by Amersham Pharmacia), and transformed into *E. coli* TG1 (manufactured by Lucigen) to obtain plasmids.

The obtained plasmid was infected with M13KO7 helper phage (manufactured by Invitrogen), thereby obtaining a human antibody M13 phage library containing the VH and VL genes.

Using the human antibody M13 phage library, an anti-rat MOG (rMOG) monoclonal antibody was obtained using the phage display method described below. The rMOG-FLAG_Fc from Example 4 (described later) was immobilized on a MAXISORP STARTUBE (manufactured by NUNC), and the unbound sites of rMOG-FLAG_Fc were blocked using SuperBlock Blocking Buffer (manufactured by Thermo).

The human antibody M13 phage library was reacted in the tube at room temperature for 1 hour. After washing with PBS or PBS containing 0.1% Tween 20 (hereinafter referred to as PBS-T), the phage was eluted with 0.1M Gly-HCl (pH 2.2). The eluent was neutralized by adding Tris-HCl (pH 8.5). The eluted phage was then used to infect TG1 competent cells to amplify the phage.

Then, the phage was reacted again with rMOG-FLAG_Fc immobilized on a MAXISORP STARTUBE, followed by washing and elution. Repeating this process yielded concentrated scFv phages that specifically bound to rMOG-FLAG_Fc. The concentrated phages were then monocloned, and three clones binding to rMOG-FLAG_Fc were selected using ELISA.

In the ELISA test, rMOG-FLAG_Fc was immobilized on a MAXISORP plate (manufactured by NUNC), and unbound sites were blocked using SuperBlock Blocking Buffer (manufactured by Thermo). As a negative control, plates immobilized with FLAG_Fc were also prepared.

Each phage clone was added to each well, and after reacting at room temperature for 30 minutes, each well was washed with PBS-T. Next, the horseradish peroxidase-labeled anti-M13 antibody (manufactured by GE Healthcare) was diluted with PBS-T containing 10% Block Ace (manufactured by Dai Nippon Pharmaceutical Co., Ltd.), and the resulting solution was added to each well and incubated at room temperature for 30 minutes.

After washing the microplate three times with PBS-T, TMB chromogenic substrate solution (DAKO) was added and incubated at room temperature. 0.5M sulfuric acid was added to each well to terminate the chromogenic reaction. The absorbance at 450 nm (reference wavelength 570 nm) was measured using a microplate reader (Molecular Devices). The results are shown in Figure 1.

As shown in Figure 1, it can be confirmed that all three phage clones bind to rMOG-FLAG Fc. On the other hand, none of the phage clones bind to FLAG Fc (data not shown).

Sequence analysis was performed on clones that bound to rMOG-FLAG_Fc to obtain the anti-MOG antibody phage vectors pCANTAB_MOG01, pCANTAB_MOG09, and pCANTAB_MOG14.

In the following text, the anti-MOG scFv antibodies displayed by phages expressed using pCANTAB_MOG01, pCANTAB_MOG09, and pCANTAB_MOG14 will be referred to as MOG01 antibody, MOG09 antibody, and MOG14 antibody, respectively. The VH or VL base sequences encoding each anti-MOG scFv antibody, and the amino acid sequences deduced from these base sequences, are shown in Table 1.

[Table 1]

Sequence information of anti-MOG scFv antibodies (MOG01 antibody, MOG09 antibody, and MOG14 antibody)

clone name MOG01 MOG09 MOG14 The base sequence encoding VH (including the signal sequence) Serial Number 1 Serial Number 13 Serial Number 25 The amino acid sequence of VH (containing the signal sequence) Serial Number 2 Serial Number 14 Serial Number 26 The amino acid sequence of VH (excluding the signal sequence) Serial Number 3 Serial Number 15 Serial Number 27 amino acid sequence of HCDR1 Serial Number 4 Serial Number 16 Serial Number 28 amino acid sequence of HCDR2 Serial Number 5 Serial Number 17 Serial Number 29 amino acid sequence of HCDR3 Serial Number 6 Serial Number 18 Serial Number 30 The base sequence encoding VL (containing the signal sequence) Serial Number 7 Serial Number 19 Serial Number 31 The amino acid sequence of VL (containing the signal sequence) Serial Number 8 Serial Number 20 Serial Number 32 The amino acid sequence of VL (excluding the signal sequence) Serial Number 9 Serial Number 21 Serial Number 33 The amino acid sequence of LCDR1 Serial Number 10 Serial Number 22 Serial Number 34 The amino acid sequence of LCDR2 Serial Number 11 Serial Number 23 Serial Number 35 The amino acid sequence of LCDR3 Serial Number 12 Serial Number 24 Serial Number 36

(2) Obtaining antibodies using alpaca antibody libraries

As an immunogen, alpacas were immunized with an emulsion of rMOG-FLAG_Fc with a complete adjuvant in the first immunization, and with an emulsion of rMOG-FLAG_Fc with an incomplete adjuvant in the second and third immunizations.

Lymphocytes (2 × ^10⁷ cells) were collected from the blood (50 mL) of immunized alpacas, and RNA was extracted from the cells using RNAIsoPlus (manufactured by TAKARA). cDNA was then synthesized via reverse transcription using the SuperScript III first-strand synthesis system (manufactured by Invitrogen) on RT-PC. The VHH gene was then amplified using primers specific to alpaca IgG2 (short hinge-heavy chain antibody) and IgG3 (long hinge-heavy chain antibody).

The VHH gene fragment was inserted into the phage vector pKSTV-02 (described in Miyazaki et al, J. Biochem. 2015; 1), and E. coli TG1 was transformed using a MicroPulser electroporator (manufactured by BioRad) (the titer of the transformant was 2.6 × ^10⁷ for IgG2 and 3.2 × ^10⁷ for IgG3).

The resulting transformants were infected with M13KO7 helper phage (manufactured by Invitrogen) to obtain an alpaca antibody M13 phage library containing the VHH gene.

Using the alpaca antibody M13 phage library, anti-MOG antibodies were obtained using the following biopanning method. rMOG-GST (4 μg/2 mL) was immobilized in an immunotherapy tube, and sites not bound to rMOG-GST were blocked using 0.5% BSA.

The alpaca antibody M13 phage library was reacted in the tube at room temperature for 1 hour. After washing with PBS-T, the phage was eluted with 0.1M Gly-HCl (pH 2.7). The eluent was neutralized by adding Tris-HCl (pH 9.1). The eluted phage was then used to infect *E. coli* TG1 to amplify the phage. The phage was then reacted again with rMOG-GST immobilized in the immunotube for washing and elution.

This operation was repeated three times for IgG2 and twice for IgG3, resulting in the concentration of phages exhibiting VHH that specifically bind to rMOG-GST. From the concentrated phages, 96 phage clones exhibiting VHH for both IgG2 and IgG3 were monocloned, and clones binding to rMOG-GST were selected using ELISA.

In the ELISA, rMOG-GST was immobilized (50 ng/50 μL) on a MAXISORP (manufactured by NUNC), and unbound sites were blocked using 0.5% BSA. Phage clones were added to each well, and after incubation at room temperature for 1 hour, each well was washed 5 times with PBS-T.

Next, 50 μL of biotinylated anti-M13 phage antibody (manufactured by Abcam) and horseradish peroxidase-labeled streptavidin (manufactured by Vector) were added to each well and incubated at room temperature for 1 hour.

After washing the microplate with PBS-T, add TMB chromogenic substrate solution (CALBIOCHEM) to each well and incubate at room temperature. Add 1M hydrochloric acid to each well to terminate the chromogenic reaction, and measure the absorbance at 450nm (reference wavelength 570nm) using a microplate reader (Model 680XR, BioRad).

Sequence analysis was performed on clones that bound rMOG-GST to obtain the anti-MOG VHH antibody iMOG-3Rim1-S32. The base sequence encoding the VHH of the iMOG-3Rim1-S32 antibody and the amino acid sequence deduced from this base sequence are shown in Table 2.

[Table 2]

Sequence information of anti-MOG VHH antibody (iMOG-3Rim1-S32 antibody)

clone name iMOG-3Rim1-S32 antibody The base sequence encoding VHH (including the signal sequence) Serial Number 37 The amino acid sequence of VHH (containing the signal sequence) Serial Number 38 The amino acid sequence of VHH (excluding the signal sequence) Serial Number 39 CDR1 amino acid sequence Serial Number 40 The amino acid sequence of CDR2 Serial Number 41 The amino acid sequence of CDR3 Serial Number 42

[Example 2] Construction of antibody expression vector

(1) Construction of expression vector for anti-MOG antibody

To produce human IgG type anti-MOG antibodies, various anti-MOG antibody expression vectors were prepared using the method described below. These vectors contained DNA sequences encoding the amino acid sequences of each variable region of an anti-MOG scFv antibody derived from the human antibody phage library obtained in Example 1.

The base sequence encoding the constant region of the λ chain of human IgG was synthesized and inserted into the BglII-EcoRI site of the N5KG4PE vector (described in International Publication No. 2002/088186) to prepare the N5LG4PE vector.

Expression vectors encoding the MOG01 and MOG09 antibodies by inserting the amino acid sequences of the VH and VL values into the N5LG4PE vector were named N5LG4PE_MOG01 and N5LG4PE_MOG09, respectively. Similarly, an expression vector encoding the MOG14 antibody by inserting the amino acid sequences of the VH and VL values into the N5KG4PE vector was named N5KG4PE_MOG14.

(1-1) MOG01 antibody expression vector N5LG4PE_MOG01

Using the phage vector pCANTAB_MOG01 as a template, the gene fragment in the VL region was amplified by PCR using primers 1 (serial number 43) and 2 (serial number 44) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.). In the PCR, 30 cycles of a reaction consisting of 94°C for 30 seconds, 58°C for 30 seconds, and 68°C for 45 seconds were performed. Unless otherwise specified, the PCR described in Example 2 was performed under the above conditions.

Using the PCR product as a template, primers 3 (serial number 45) and 2 (serial number 44) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.) were used to attach a signal sequence to the gene fragment in the VL region by PCR.

The obtained gene fragment was inserted into the BglII-BlpI site of the N5LG4PE vector to obtain N5LG4PE_MOG01VL. Next, using pCANTAB_MOG01 as a template, the gene fragment in the VH region was amplified by PCR using primers 4 (serial number 46) and 5 (serial number 47) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

Using the PCR product as a template, a signal sequence was added to the gene fragment in the VH region by PCR using primers 6 (serial number 48) and 5 (serial number 47) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.). The obtained gene fragment was then inserted into the SalI-NheI site of the N5LG4PE_MOG01VL vector to obtain N5LG4PE_MOG01.

(1-2) MOG09 antibody expression vector N5LG4PE_MOG09

N5LG4PE_MOG09 was prepared using the same method as described in (1-1) above. Using the phage vector pCANTAB_MOG09 as a template, primers 7 (SEQ ID NO: 49) and 8 (SEQ ID NO: 50) were used to amplify the gene fragment in the VL region. Primers 3 (SEQ ID NO: 45) and 8 (SEQ ID NO: 50) were used to attach the signal sequence to the gene fragment in the VL region. Primers 9 (SEQ ID NO: 51) and 10 (SEQ ID NO: 52) were used to amplify the gene fragment in the VH region. Primers 6 (SEQ ID NO: 48) and 10 (SEQ ID NO: 52) were used to attach the signal sequence to the gene fragment in the VH region.

(1-3) MOG14 antibody expression vector N5KG4PE_MOG14

N5KG4PE_MOG14 was prepared using the same method as described in (1-1). Using the phage vector pCANTAB_MOG14 as a template, primers 11 (SEQ ID NO. 53) and 12 (SEQ ID NO. 54) were used to amplify the gene fragment in the VL region. Primers 3 (SEQ ID NO. 45) and 12 (SEQ ID NO. 54) were used to attach the signal sequence to the gene fragment in the VL region. The resulting gene fragment in the VL region with the attached signal sequence was inserted into the BglII-BsiWI site of the N5KG4PE vector to obtain N5KG4PE_MOG14VL.

Next, using pCANTAB_MOG14 as a template, primers 13 (SEQ ID NO: 55) and 14 (SEQ ID NO: 56) were used to amplify the gene fragment in the VH region. Primers 6 (SEQ ID NO: 48) and 14 (SEQ ID NO: 56) were used to attach the signal sequence to the gene fragment in the VH region. The resulting gene fragment in the VH region with the attached signal sequence was inserted into the SalI-NheI site of N5KG4PE_MOG14VL, yielding N5KG4PE_MOG14.

(1-4) iMOG-3Rim1-S32 antibody expression vector N5G4PEFc_iMOG-3Rim1-S32

A sequence with a signal sequence attached to the gene encoding the Fc region of human IgG4PE was synthesized, and the gene fragment of the human Fc region was amplified by PCR using primers 25 (serial number 79) and 26 (serial number 80) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, a reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 60 seconds at 68°C was performed for 30 cycles. The resulting Fc gene fragment was inserted into the BglII-BamHI site of the N5KG4PE vector to create the N5G4PEFc vector.

The expression vector, named N5G4PEFc_iMOG-3Rim1-S32, was created by inserting the amino acid sequence encoding VHH (iMOG-3Rim1-S32) into N5G4PEFc. The VHH-Fc expression vector was prepared using the method described below.

The VHH base sequence of iMOG-3Rim1-S32 was synthesized. The VHH gene fragment was amplified by PCR using primers 15 (SEQ ID NO: 57) and 16 (SEQ ID NO: 58) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.). The PCR reaction consisted of 30 cycles of 94°C for 30 seconds, 58°C for 30 seconds, and 68°C for 60 seconds. The obtained VHH gene fragment was inserted into the EcoRI-BglII site of the N5G4PEFc vector to obtain N5G4PEFc_iMOG-3Rim1-S32.

(2) Expression vector for anti-ivemectin antibody N5LG4PE_AVM

A chimeric anti-avermectin (AVM) antibody was prepared as a negative control antibody using the same method as described in (1-1). The expression vector formed by inserting the base sequences encoding the amino acid sequences VH and VL of the AVM antibody into N5LG4PE was named N5LG4PE_AVM.

AVMs were immunized in SD rats, and anti-AVM antibodies were generated to produce hybridomas using standard methods. The variable region of the anti-AVM antibody derived from this hybridoma was used as a template. Primers 29 (SEQ ID NO: 83) and 30 (SEQ ID NO: 84) were used to amplify the gene fragment in the VL region, while primers 3 (SEQ ID NO: 45) and 30 (SEQ ID NO: 84) were used to attach a signal sequence to the gene fragment in the VL region.

To amplify the gene fragment in the VH region, primers 31 (serial number 85) and 32 (serial number 86) were used. When attaching the signal sequence to the gene fragment in the VH region, primers 6 (serial number 48) and 32 (serial number 86) were used.

(3) Expression vector for anti-rat transferrin receptor antibody OX26: N5KG4PE(R409K)_OX26

As a positive control antibody against rat transferrin receptor antibody, the OX26 antibody against rat transferrin receptor antibody described in [Protein Engineering, 12, 787-796, 1999] was prepared. An expression vector encoding the VH and VL amino acid sequences of the OX26 antibody was prepared by inserting the base sequence of the OX26 antibody into N5KG4PE(R409K) (described in International Publication No. 2002/088186) using the same method as described in (1-1). This vector was named N5KG4PE(R409K)_OX26.

The gene encoding the amino acid sequence of VL of the OX26 antibody was used as a template. Primers 40 (Sequence No. 94) and 41 (Sequence No. 95) were used to amplify the gene fragment in the VL region, and primers 42 (Sequence No. 96) and 43 (Sequence No. 97) were used to amplify the gene fragment in the VH region.

[Example 3] Construction of expression vector for bispecific antibodies

(1) Preparation of expression vectors for bispecific antibodies that bind to Her2 and MOG

The expression vector pCI-Trastuzumab-hKG4PE(R409K)_MOG01scFv, which is a bispecific antibody binding to HER2 and MOG, was prepared using the following method. This bispecific antibody is formed by fusing the scFv of an anti-MOG antibody to the C-terminus of both H chains of an anti-HER2 antibody IgG.

Using the synthetic gene of the heavy chain constant region as a template, the gene fragment of the CH1-hinge-CH2-CH3-connector region was amplified by PCR using primers 17 (serial number 59) and 18 (serial number 60) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, the reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 2 minutes at 68°C was performed for 30 cycles. Using the phage vector pCANTAB_MOG01 as a template, primers 19 (serial number 61) and 20 (serial number 62) and KODplus DNA polymerase (manufactured by Toyobo Co., Ltd.) were used to amplify the gene fragment of the scFv region (hereinafter referred to as MOG01scFv) by PCR.

In the PCR, the reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 90 seconds at 68°C was performed for 30 cycles. Then, using the CH1-hinge-CH2-CH3 region and the MOG01scFv region described above as templates, the CH1-hinge-CH2-CH3-MOG01scFv region was amplified using primers 17 (SEQ ID NO. 59) and 20 (SEQ ID NO. 62) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, a reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 2 minutes at 68°C was performed for 30 cycles. The resulting gene fragment was inserted into a pCI vector (manufactured by Promega) to create the pCI-hG4PE(R409K)_MOG01scFv vector.

Using the gene encoding the amino acid sequence of VL of the anti-HER2 antibody (trastuzumab) (described in International Publication No. 1999/57134) as a template, the gene fragment of the VL region was amplified by PCR using primers 21 (Sequence No. 63) and 22 (Sequence No. 64) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, the reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 45 seconds at 68°C was performed for 30 cycles. Using the N5KG4PE vector (described in International Publication No. 2002/088186) as a template, primers 27 (Sequence No. 81) and 28 (Sequence No. 82) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.) were used to amplify the gene fragment in the CL region by PCR.

In the PCR, the reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 45 seconds at 68°C was performed for 30 cycles. Using the obtained gene fragments VL and CL as templates, the gene fragments were amplified by PCR using primers 21 (serial number 63) and 28 (serial number 82) and KODplus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, a reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 90 seconds at 68°C was performed for 30 cycles. The resulting gene fragment was inserted into pCI-hG4PE(R409K)_MOG01scFv to obtain pCI-TrastuzumabVL-hKG4PE(R409K)_MOG01scFv.

Next, using the gene encoding the amino acid sequence of VH of trastuzumab as a template, the gene fragment of the VH region was amplified by PCR using primers 23 (SEQ ID NO: 65) and 24 (SEQ ID NO: 66) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.). The PCR reaction consisted of 30 cycles of 94°C for 30 seconds, 58°C for 30 seconds, and 68°C for 45 seconds.

The obtained gene fragment was inserted into pCI-TrastuzumabVL-hKG4PE(R409K)_MOG01scFv to obtain pCI-Trastuzumab-hKG4PE(R409K)_MOG01scFv.

(2) Preparation of expression vectors for bispecific antibodies binding to AVM and MOG

In addition, the expression vector pCI-AVM-hLG4PE(R409K)_MOG01scFv, which contains a bispecific antibody binding to AVM and MOG, was prepared using the method described below. This bispecific antibody was prepared by fusing an anti-MOG antibody scFv to the C-terminus of an anti-AVM antibody IgG.

Using N5LG4PE_AVM as a template, the gene fragment of the light chain region of AVM was amplified by PCR using primers 33 (serial number 87) and 34 (serial number 88) and KODplus DNA polymerase (manufactured by Toyobo Co., Ltd.). The PCR reaction consisted of 30 cycles of 94°C for 30 seconds, 58°C for 30 seconds, and 68°C for 60 seconds.

Using N5LG4PE_AVM as a template, the gene fragment of the AVM VH region was amplified by PCR using primers 35 (serial number 89) and 32 (serial number 86) and KODplus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, a reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 45 seconds at 68°C was performed for 30 cycles. The resulting gene fragment was inserted into the prepared pCI-hG4PE(R409K)_MOG01scFv to obtain pCI-AVM-hLG4PE(R409K)_MOG01scFv.

(3) Preparation of an antibody expression vector by fusing an anti-AVM antibody scFv to the C-terminus of an anti-AVM antibody IgG.

The expression vector of the antibody, which is formed by fusing the anti-AVM antibody scFv to the C-terminus of the anti-AVM antibody IgG as a negative control antibody, is named pCI-AVM-hLG4PE(R409K)_AVM scFv.

Using the synthetic gene of the heavy chain constant region as a template, the gene fragment of the CH1-hinge-CH2-CH3-connector region was amplified by PCR using primers 36 (serial number 90) and 37 (serial number 91) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, 30 cycles of a reaction consisting of 30 seconds at 94°C, 30 seconds at 58°C, and 2 minutes at 68°C were performed. Using the synthetic gene of AVM scFv as a template, the gene fragment of the scFv region was amplified by PCR using primers 38 (serial number 92) and 39 (serial number 93) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.).

In the PCR, 30 cycles of a reaction consisting of 94°C for 30 seconds, 58°C for 30 seconds, and 68°C for 90 seconds were performed. Next, using the CH1-hinge-CH2-CH3 region and the AVM scFv region described above as templates, primers 36 (SEQ ID NO: 90) and 39 (SEQ ID NO: 93) and KOD plus DNA polymerase (manufactured by Toyobo Co., Ltd.) were used to amplify the CH1-hinge-CH2-CH3-AVM scFv region. In the PCR, 30 cycles of a reaction consisting of 94°C for 30 seconds, 58°C for 30 seconds, and 68°C for 2 minutes were performed.

The obtained gene fragment was inserted into the NheI-BamHI site of the above pCI-AVM-hLG4PE(R409K)_MOG01scFv to obtain pCI-AVM-hLG4PE(R409K)_AVM scFv.

[Example 4]

Preparation of soluble MOG antigen and soluble HER2 antigen

(1) Preparation of extracellular domain proteins of FLAG-Fc-binding rat MOG

The extracellular domain protein of the rat MOG, with FLAG-Fc appended to the C-terminus, was prepared using the method described below. The base sequence encoding rMOG is shown in Serial No. 67, and the amino acid sequence deduced from this base sequence is shown in Serial No. 68.

The gene sequence for the extracellular domain of MOG was synthesized and inserted into the BglII-XbaI site of the INPEP4 (manufactured by IDEC) vector containing FLAG-Fc, thereby creating the plasmid vector INPEP4_rMOG-FLAG-Fc expressing the extracellular domain of MOG with FLAG-Fc appended to the C-terminus. The base sequence of rMOG-FLAG-Fc is shown in SEQ ID NO: 69, and the amino acid sequence deduced from this base sequence is shown in SEQ ID NO: 70.

INPEP4_rMOG-FLAG-Fc was introduced into suspension 293 cells using the Expi293 (trademarked) expression system (manufactured by Thermo Fisher Scientific) and cultured to express the protein in a transient expression system. Four days after vector introduction, the culture supernatant was collected and filtered through a 0.22 μm membrane filter (manufactured by MillIPORE).

The MOG-FLAG-Fc protein in the culture supernatant was purified by affinity using protein A resin (MabSelect SuRe, manufactured by GE Healthcare Bioscience). Phosphate buffer was used as the washing solution.

The protein adsorbed on protein A was eluted using 20 mM sodium citrate and 50 mM NaCl buffer (pH 3.4) and recovered into a tube containing 1 M Tris-HCl buffer (pH 8.0).

Next, the solvent in the eluent was replaced with PBS using ultrafiltration with a VIVASPIN (Sartrius Stealin) and a NAP column (GE Healthcare Bioscience), followed by filtration and sterilization using a 0.22 μm membrane filter (Millex-GV, MillIPORE). The concentrations of the purified MOG-FLAG-Fc proteins in the solution were determined by absorbance at 280 nm.

(2) Production of GST-binding MOG extracellular domain proteins

The extracellular domain protein of the rat MOG, with GST appended to the C-terminus, was prepared using the method described below.

The gene sequence for the extracellular domain of MOG was synthesized and inserted into the BglII-KpnI site of an N5 vector (manufactured by IDEC) containing GST, thereby creating the plasmid vector N5_rMOG-GST expressing the extracellular domain of MOG with GST attached to the C-terminus. The base sequence of rMOG-GST is shown in Serial No. 71, and the amino acid sequence deduced from this base sequence is shown in Serial No. 72.

The extracellular domain protein of HER2, which is a soluble antigen of human HER2 with a GST appended to its C-terminus, was prepared using the method described below. The gene sequence of the extracellular domain of HER2 was synthesized and inserted into the BglII-KpnI site of an N5 vector (manufactured by IDEC) containing a GST appended, thereby creating the plasmid vector N5_hHER2-GST expressing the extracellular domain of HER2 with a GST appended to its C-terminus. The base sequence of hHER2-GST is shown in Serial No. 71, and the amino acid sequence deduced from this base sequence is shown in Serial No. 72.

Using the Expi293 (trademarked) expression system (manufactured by Thermo Fisher Scientific), N5_rMOG-GST and N5_hHER2-GST were introduced into suspension 293 cells and cultured for protein expression in a transient expression system. Four days after vector introduction, the culture supernatant was collected and filtered through a 0.22 μm membrane filter (manufactured by MillIPORE).

Proteins in the culture supernatant were purified by affinity using glutathione agarose 4B (manufactured by GE Healthcare Bioscience). Phosphate buffer was used as the washing solution. Proteins adsorbed on glutathione agarose 4B were eluted with 50 mM Tris-HCl and 10 mM reduced glutathione (pH 8.0).

Next, after replacing the solvent in the solution with PBS using ultrafiltration with a VIVASPIN (Sartrius Stealin) and an NAP column (GE Healthcare Bioscience), the solution was sterilized by filtration through a 0.22 μm membrane filter (Millex-GV, MillIPORE). The concentrations of the purified rMOG-GST and hHER2-GST proteins in the solution were determined by absorbance at 280 nm.

[Example 5] Preparation of membrane-type MOG antigen expression vector

The full-length gene sequences of rat MOG (rMOG), mouse MOG (mMOG), monkey MOG (cMOG), and human MOG (hMOG) were synthesized. Each gene sequence was inserted into the BamHI-NotI site of the pEF6/V5-His vector (manufactured by Thermo Fisher Scientific) to create membrane expression plasmid vectors for various MOGs, namely pEF6_rMOG, pEF6_mMOG, pEF6_cMOG, and pEF6_hMOG.

The base sequence encoding mMOG is shown in Serial No. 73, and the amino acid sequence deduced from this base sequence is shown in Serial No. 74. The base sequence encoding cMOG is shown in Serial No. 75, and the amino acid sequence deduced from this base sequence is shown in Serial No. 76. The base sequence encoding hMOG is shown in Serial No. 77, and the amino acid sequence deduced from this base sequence is shown in Serial No. 78.

[Example 6] Preparation of various antibodies

The antibody expression plasmid vectors prepared in Examples 2 and 3 were introduced into suspension 293 cells and cultured using the Expi293 ^™ expression system (manufactured by Thermo Fisher Scientific) to express the antibodies in a transient expression system.

Four days after vector introduction, the culture supernatant was recovered and filtered using a 0.22 μm membrane filter (MILLIPORE). Protein A resin (MabSelect SuRe, GE Healthcare Bioscience) was used for affinity purification of the protein in the culture supernatant. Phosphate buffer was used as the washing buffer. The antibody adsorbed onto protein A was eluted using 20 mM sodium citrate and 50 mM NaCl buffer (pH 3.4) and recovered into a tube containing 1 M Tris-HCl buffer (pH 8.0).

Next, the solvent of the eluent was replaced with PBS using ultrafiltration with a VIVASPIN (Sartrius Stealin) and a NAP column (GE Healthcare Bioscience), followed by filtration and sterilization using a 0.22 μm membrane filter (Millex-GV, MillIPORE). The absorbance of the antibody solution at 280 nm was measured, and the concentration of 1 mg/mL was converted to an optical density of 1.40 to calculate the concentration of the purified antibody.

The anti-MOG human IgG antibodies expressed using the expression vectors N5LG4PE_MOG01, N5LG4PE_MOG09, N5KG4PE_MOG14, and N5G4PEFc_iMOG-3Rim1-S32 described in Example 2 are respectively labeled as MOG01 antibody, MOG09 antibody, MOG14 antibody, and iMOG-3Rim1-S32 antibody.

The antibodies obtained by expressing the various bispecific antibody expression vectors pCI-AVM-hLG4PE(R409K)_AVMscFv, pCI-AVM-hLG4PE(R409K)_MOG01scFv and pCI-Trastuzumab-hKG4PE(R409K)_MOG01scFv prepared in Example 3 were named AVM IgG4PE(R409K)_AVM dscFv antibody, AVM IgG4PE(R409K)_MOG01dscFv antibody and Trastuzumab IgG4PE(R409K)_MOG01scFv antibody, respectively.

[Example 7] Evaluation of the binding affinity of anti-MOG antibodies to MOG using flow cytometry

The binding of the anti-MOG antibodies MOG01, MOG09, MOG14 and iMOG-3Rim1-S32 obtained in Example 6 to MOG was evaluated using fluorescence activated cell sorting (FACS) following the steps described below.

The various membrane-type MOG antigen expression vectors prepared in Example 5 were introduced into suspension 293 cells and cultured using the FreeStyle (trademark) 293 expression system (manufactured by Thermo Fisher Scientific), allowing the membrane-type antigens to be expressed in a transient expression system. Using these cells, the reactivity to anti-MOG antibodies was analyzed using the methods described below.

rMOG/HEK293F, mMOG/HEK293F, cMOG/HEK293, and hMOG/HEK293 cells were suspended at a concentration of 5 × ^10⁵ cells/mL in staining buffer (SB) containing 0.1% _NaN³⁺ and 1% FBS, and then dispensed into 96-well round-bottom plates (manufactured by BD).

After centrifugation (2000 rpm, 4°C, 2 min), the supernatant was removed, and 10 μg/mL of each antibody obtained in Example 6 was added to the clump and the mixture was resuspended. The mixture was then incubated at ice temperature for 30 min. Further centrifugation (2000 rpm, 4°C, 2 min) was performed, the supernatant was removed, and the clump was washed with SB. Then, 1 μg/mL of RPE fluorescently labeled goat anti-human antibody (manufactured by Southern Bioblot) was added, and the mixture was incubated at ice temperature for 30 min.

After washing with SB, the cells were resuspended in SB, and the fluorescence intensity of each cell was measured using a flow cytometer FACS CANTO II (manufactured by BD). The results are shown in Figure 2. It should be noted that an anti-AVM antibody was used as a negative control.

As shown in Figure 2, the MOG01, MOG09, MOG14, and iMOG-3Rim1-S32 antibodies, acting as anti-MOG antibodies, all exhibited binding activity against both rMOG/HEK293F and mMOG/HEK293F cells. Furthermore, the MOG01 and MOG14 antibodies also showed binding activity against both cMOG/HEK293 and hMOG/HEK293 cells.

This indicates that the anti-MOG human IgG antibodies MOG01 and MOG14 not only recognize and bind to MOGs in rats and mice, but also recognize and bind to all MOGs in cynomolgus monkeys and humans.

[Example 8] Evaluation of the binding affinity of anti-MOG antibodies to MOG using surface plasmon resonance detection

The affinity of the anti-MOG antibodies MOG01, MOG09, MOG14 and iMOG-3Rim1-S32 obtained in Example 6 for rat MOG was determined using a Biacore T-100 (GE Healthcare).

Antibodies were immobilized on the CM5 sensor chip using a human antibody capture kit, and the binding affinity was evaluated using rMOG-GST prepared in Example 4 as the analyte. The obtained sensor maps were analyzed using BIA evaluation software to calculate the dissociation constant (KD value). The results are shown in Table 3.

[Table 3]

As shown in Table 3, the dissociation constants (KD values) of each anti-MOG antibody range from 2.1 × ^10¹¹ (M) to 4.0 × ^10⁻⁸ (M), indicating that all antibodies exhibit good affinity. For the MOG09 antibody, the dissociation rate constant kd is outside the measurement range of the equipment, and the KD value cannot be uniquely determined.

[Example 9] Evaluation of brain migration of anti-MOG antibody in rats

After administering the antibody via tail vein (i.v.) to rats, blood was collected from the tail vein. On the same day as blood collection, the rats underwent systemic perfusion under pentobarbital anesthesia, and brain tissue was recovered and its weight measured. Additionally, a buffer solution was added to the recovered brain tissue, homogenized, and centrifuged. The antibody solution dissolved in the supernatant was recovered. Its volume was measured, and the antibody concentration was determined using an AlphaLISA (manufactured by PerkinElmer) to calculate the amount of antibody per unit brain weight.

For the anti-MOG antibodies MOG01, MOG14, iMOG-3Rim1-S32, and the negative control anti-AVM antibody, the MOG01 and MOG14 antibodies were administered at a dose of 1 mg/kg body weight, and the iMOG-3Rim1-S32 antibody was administered at a dose of 5 mg/kg body weight. The antibody concentration in serum and the amount of antibody per unit brain weight in brain tissue after 4 days are shown in Figures 3(A) and (B).

As shown in Figures 3(A) and (B), the serum antibody concentration of any one of the anti-MOG antibodies did not change compared to the negative control (AVM), but the amount of antibodies in the brain increased by 5-10 times.

Anti-MOG antibody MOG01 antibody, anti-transferrin receptor antibody OX26 antibody, and negative control anti-AVM antibody were administered at a dose of 5 mg/kg body weight. The serum antibody concentrations and the amount of antibody per unit brain weight in brain tissue after 4 and 10 days are shown in Figures 4(A) and (B).

As shown in Figure 4(A), with the OX26 antibody, the serum antibody concentration was the lowest among the evaluated antibodies after 4 days, and below the detection sensitivity after 10 days, indicating poor serum antibody dynamics. With the MOG01 antibody, an anti-MOG antibody, there were no significant changes in serum antibody concentration after 4 and 10 days of administration, remaining at the same level as the negative control. Therefore, it can be said that the serum half-life of the MOG01 antibody is similar to that of the negative control.

In addition, as shown in Figure 4(B), regarding the amount of antibodies in the brain, the negative control had the lowest antibody level among the evaluated antibodies 4 days after administration, and the antibody level further decreased slightly after 10 days.

The antibody levels of OX26 decreased rapidly between 4 and 10 days after administration, and the antibody levels after 10 days of administration were below those of the negative control. In contrast, the antibody levels of MOG01 increased between 4 and 10 days after administration, with the antibody levels after 4 days of administration reaching approximately 2.5 times that of the negative control and after 10 days of administration reaching approximately 10 times that of the negative control.

As shown above, the anti-MOG antibody MOG01 showed the same antibody concentration in serum as the negative control, but the antibody level in the brain increased to about 2.5 times that of the negative control after 4 days of administration and to about 10 times that of the negative control and OX26 antibody after 10 days of administration.

[Example 10] Evaluation of the binding affinity of bispecific antibodies to MOG or HER2 using flow cytometry

The binding of the bispecific antibodies Trastuzumab IgG4PE(R409K)_MOG01dscFv (binding to MOG and Her2), AVM IgG4PE(R409K)_MOG01dscFv (binding to MOG and AVM), and AVM IgG4PE(R409K)_AVM dscFv (binding to AVM) obtained in Example 6 to MOG or HER2 was evaluated using fluorescence activated cell sorting (FACS) as described below.

The membrane-type MOG antigen expression vector prepared in Example 5 was introduced into suspension 293 cells and cultured using the FreeStyle 293 expression system (manufactured by Thermo Fisher Scientific) to express the membrane-type antigen in a transient expression system.

HEK293F cells, rMOG/HEK293F cells, hMOG/HEK293F cells, and human breast cancer cell line SK-BR-3 cells were suspended at a concentration of 5 × ^10⁵ cells/mL in staining buffer (SB) containing 0.1% _NaN₃ and 1% FBS, and then dispensed into 96-well round-bottom plates (manufactured by BD).

After centrifugation (2000 rpm, 4°C, 2 min), the supernatant was removed, and 10 μg/mL of each antibody obtained in Example 6 was added to the clump and the mixture was resuspended. The mixture was then incubated at ice temperature for 30 min. Further centrifugation (2000 rpm, 4°C, 2 min) was performed, the supernatant was removed, and the clump was washed with SB. Then, 1 μg/mL of RPE fluorescently labeled goat anti-human antibody (manufactured by Southern Bioblot) was added, and the mixture was incubated at ice temperature for 30 min.

After washing with SB, the cells were resuspended in SB, and the fluorescence intensity of each cell was measured using a flow cytometer FACS CANTO II (manufactured by BD). It should be noted that 10 μg/mL of anti-AVM antibody was used as a negative control. The results of binding analysis on HEK293F cells, rMOG/HEK293F cells, and hMOG/HEK293 cells are shown in Figure 5.

As shown in Figure 5, binding to rMOG/HEK293F cells and hMOG/HEK293 cells was observed for AVM IgG4PE(R409K)_MOG01dscFv antibody and Trastuzumab IgG4PE(R409K)_MOG01dscFv antibody, thus demonstrating that even in the form of bispecific antibodies, binding to rat MOG and human MOG is maintained.

The results of binding analysis on the human breast cancer cell line SK-BR-3 are shown in Figure 6. HER2 is known to be expressed in this cell line.

As shown in Figure 6, the Trastuzumab IgG4PE(R409K)_MOG01dscFv antibody, even in the form of a bispecific antibody, maintains its binding to HER2.

[Example 11] Evaluation of MOG binding affinity of bispecific antibodies to MOG using surface plasmon resonance detection

The affinity of the bispecific antibody for MOG was determined using the same method as in Example 8, and the results are shown in Table 4.

[Table 4]

As shown in Table 4, the dissociation constant (KD value) of each bispecific antibody is 2.0 × ^10⁻⁷ (M) for AVM IgG4PE(R409K)_MOG01dscFv antibody and 1.0 × ^10⁻⁷ (M) for Trastuzumab IgG4PE(R409K)_MOG01dscFv antibody, indicating that bispecific antibodies of any MOG exhibit good affinity.

In the case of Trastuzumab IgG4PE(R409K)_MOG01dscFv antibody, the binding rate constant ka and dissociation rate constant kd are outside the measurement range of the device, and the KD value cannot be uniquely determined.

[Example 12] Evaluation of the binding affinity of MOG bispecific antibodies to HER2 using surface plasmon resonance detection

The affinity of the bispecific antibody Trastuzumab IgG4PE(R409K)_MOG01dscFv for HER2 was determined using a Biacore T-100 (GE Healthcare).

Antibodies were immobilized on the CM5 sensor chip using a human antibody capture kit. The binding affinity of the MOG-Her2 bispecific antibody was evaluated using HER2-GST prepared in Example 4 as the analyte. The obtained sensor maps were analyzed using BIA evaluation software to calculate the dissociation constant (KD value). The results are shown in Table 5.

[Table 5]

As shown in Table 5, the dissociation constant (KD value) of Trastuzumab IgG4PE(R409K)_MOG01dscFv antibody against HER2 is 3.7× ^10⁻⁹ (M), indicating that it is an antibody with good affinity.

[Example 13] Evaluation of the brain migration of MOG bispecific antibodies in rats

The brain migration of the bispecific antibodies AVM IgG4PE(R409K)_MOG01dscFv, Trastuzumab IgG4PE(R409K)_MOG01dscFv, and AVM IgG4PE(R409K)_AVM dscFv in rats was evaluated using the same method as in Example 9. Antibodies were administered at a dose of 5 mg/kg body weight, and the serum antibody concentration and antibody amount per unit brain weight after 10 days are shown in Figures 7(A) and (B).

As shown in Figure 7(A), the serum antibody concentrations of AVM IgG4PE(R409K)_MOG01dscFv antibody and Trastuzumab IgG4PE(R409K)_MOG01dscFv antibody were not different from those of AVM IgG4PE(R409K)_AVM dscFv antibody, which served as a negative control for bispecific antibodies.

On the other hand, as shown in Figure 7(B), the amount of antibodies in the brain was increased by about 10 times compared with the AVM IgG4PE(R409K)_AVM dscFv antibody, which served as a negative control for bispecific antibodies, with the AVM IgG4PE(R409K)_AVM dscFv antibody.

As shown above, compared with bispecific antibodies that do not bind to MOG, bispecific antibodies that bind to MOG can increase the amount of antibodies in the brain by about 10 times, while the blood half-life remains unchanged.

[Example 14] Evaluation of mouse brain migration of anti-MOG01 antibody

(1) Antibody level determination

Mice were administered the antibody via tail vein (i.v.) at a dose of 35 nmol/kg, and blood was collected from the tail vein several days later. On the same day as blood collection, the brain tissue was perfused under pentobarbital anesthesia, and its weight was measured. Buffer solution was added to the recovered brain tissue, homogenized, and centrifuged. The antibody solution dissolved in the supernatant was recovered. Its volume was measured, and the antibody concentration was determined using an AlphaLISA (PerkinElmer) to calculate the amount of antibody per unit brain weight.

The serum antibody concentration and the amount of antibody per unit brain weight in brain tissue after 3, 6, 10, 14, 21, and 28 days following antibody administration of the anti-MOG01 human IgG antibody and the negative control anti-AVM human IgG antibody are shown in Figure 8(A) and (B), respectively.

As shown in Figure 8(A), the serum antibody concentration of anti-MOG01 human IgG antibody was not different from that of the negative control. On the other hand, as shown in Figure 8(B), it was shown that the antibody level in the brain increased by several tens of times over a 28-day period.

(2) Imaging analysis

Anti-MOG01 human IgG antibody and negative control anti-AVM human IgG antibody were labeled using the Alexa FluorR 488 protein labeling kit (manufactured by Molecular Probes). The labeled antibodies were designated as AF488-MOG01IgG4PE antibody and AF488-AVMIgG4PE antibody, respectively.

The labeled antibody was administered to mice via tail vein (i.v.) at a dose of 10 mg/kg. Several days later, tomato lectin was administered, and blood was collected from the cheek. After blood collection, the mice were anesthetized with pentobarbital and then perfused systemically. Brain tissue was then recovered, and fluorescence intensity was measured using IVIS Spectrum (PerkinElmer). Brain imaging images after 6 days are shown in Figure 9(A), and brain imaging images after 14 days are shown in Figure 9(B). The amount of fluorescence in the brain after correction for the fluorescence intensity of the administered antibody is shown in Figure 9(C).

As shown in Figures 9(A) to (C), the amount of anti-MOG01 antibody was increased by tens of times throughout the brain compared to the negative control.

[Example 15] Construction of various bispecific antibody expression vectors

Bispecific antibody expression vectors binding to AVM and MOG, having the structures shown in Figures 10(A)–(C) and Figures 11(A) and (B), were prepared using the method described below. The names of the bispecific antibodies and the antibody expression vectors are shown in Table 6, and the names of the antibody expression vectors, the base sequences of the antibodies, and the amino acid sequences deduced from the base sequences are shown in Table 7.

[Table 6]

[Table 7]

(1) Construction of bispecific antibody expression vector related to the structure in Figure 10(A)

(1-1) Construction of pCI-AVM-hLG4PE(R409K/S354C/T366W)-FLAG tag carrier

Using the synthesized gene as a template, the gene fragment in the CH1-hinge-CH2-CH3 (R409K/S354C/T366W) region was amplified by PCR and inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVMscFv to create the pCI-AVM-hLG4PE(R409K)(R354C/T366W)-FLAG tag vector.

(1-2) Construction of pCI-MOG01-hLG4PE(R409K/Y349C/T366S/L368A/Y407V)-His tag vector

Using the synthesized gene as a template, the gene fragment of the CH1-hinge-CH2-CH3(R409K/Y349C/T366S/L368A/Y407V)-His tag region was amplified by PCR. Additionally, using N5LG4PE_MOG01 as a template, the gene fragments of the MOG01 light chain region and the MOG01VH region were amplified by PCR. The resulting gene fragments were inserted into a pCI vector (manufactured by Promega) to create the pCI-MOG01-hLG4PE(R409K/Y349C/T366S/L368A/Y407V)-His tag vector.

(2) Construction of bispecific antibody expression vector related to the structure in Figure 10(B)

(2-1) Construction of pCI-AVM-hLG4PE(R409K)-connector-MOG01VL-CL carrier

Using the synthesized gene as a template, the gene fragment in the CH1-hinge-CH2-CH3-linker-MOG01VL-CL region was amplified by PCR and inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVMscFv to create the pCI-AVM-hLG4PE(R409K)-linker-MOG01VL-CL vector.

(2-2) Construction of pCI-MOG01VH-CH vector

Using the synthetic gene as a template, the gene fragment of the MOG01VH-CH region was amplified by PCR and inserted into the pCI vector (manufactured by Promega) to create the pCI-MOG01VH-CH vector.

(3) Construction of bispecific antibody expression vector related to the structure in Figure 10(C)

(3-1) Construction of pCI-AVM-hLG4PE(R409K/S354C/T366W)-Connector-MOG01scFv-FLAG Tag Carrier

Using the synthesized gene as a template, the gene fragment of the CH1-hinge-CH2-CH3(R409K/S354C/T366W)-linker region was amplified by PCR. Additionally, using MOG01scFv as a template, the gene fragment of the linker-MOG01scFv region was amplified by PCR. Then, using the PCR product as a template, the gene fragment of the linker-MOG01scFv-FLAG tag region was amplified by PCR. The gene fragments of the CH1-hinge-CH2-CH3(R409K/S354C/T366W)-linker region and the linker-MOG01scFv-FLAG tag region were inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVM scFv to create the pCI-AVM-hLG4PE(R409K/S354C/T366W)-linker-MOG01scFv-FLAG tag vector.

(3-2) Construction of pCI-AVM-hLG4PE(R409K/Y349C/T366S/L368A/Y407V)-Hi tag vector

Using the synthesized gene as a template, the gene fragment of the CH1-hinge-CH2-CH3(R409K/Y349C/T366S/L368A/Y407V)-His tag region was amplified by PCR and inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVMscFv to create the pCI-AVM-hLG4PE(R409K/Y349C/T366S/L368A/Y407V)-His tag vector.

(4) Construction of bispecific antibody expression vectors with the structures shown in Figure 11(A) and (B)

(4-1) Construction of the pCI-AVM-hLG4PE(R409K)_MOG01scFv vector

Using the synthesized gene as a template, the gene fragment of the CH1-hinge-CH2-CH3-connector region was amplified by PCR. Additionally, using MOG01scFv as a template, the gene fragments of the VH and VL regions of MOG01 were amplified by PCR. The gene fragments of the CH1-hinge-CH2-CH3-connector region and the gene fragments of the VH and VL regions of MOG01 were inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVMscFv to create the pCI-AVM-hLG4PE(R409K)_MOG01scFv2 vector.

Similarly, the following vectors were prepared: pCI-AVM-hLG4PE(R409K)_MOG01scFv3, pCI-AVM-hLG4PE(R409K)_MOG01scFv4, pCI-AVM-hLG4PE(R409K)_MOG01scFv5, pCI-AVM-hLG4PE(R409K)_MOG01scFv6, and pCI-AVM-hLG4PE(R409K)_MOG01scFv6. The vectors pCI-AVM-hLG4PE(R409K)_MOG01scFv7, pCI-AVM-hLG4PE(R409K)_MOG01scFv8, pCI-AVM-hLG4PE(R409K)_MOG01scFv9, pCI-AVM-hLG4PE(R409K)_MOG01scFv10, and pCI-AVM-hLG4PE(R409K)_MOG01scFv11 are listed.

(5) Construction of the expression vector for the antibody as a negative control

An antibody was prepared as a negative control using the method described below. The name of the antibody and the name of the antibody expression vector are shown in Table 8, and the name of the antibody expression vector, the base sequence of the antibody, and the amino acid sequence deduced from the base sequence are shown in Table 9.

[Table 8]

[Table 9]

(2-1) Fabrication of pCI-AVM-hLG4PE(R409K) vector

Using the synthetic gene as a template, gene fragments of the VH region, VL region, and antibody constant region of AVM were amplified by PCR and inserted into the pCI vector (manufactured by Promega) to create the pCI-AVM-hLG4PE(R409K) vector.

(2-2) Fabrication of pCI-AVM-hLG4PE(R409K)-Connector-AVMVL-CL Carrier

Using the synthesized gene as a template, the gene fragment in the CH1-hinge-CH2-CH3-linker-AVMVL-CL region was amplified by PCR and inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVM scFv to create the pCI-AVM-hLG4PE(R409K)-linker-AVMVL-CL vector.

(2-3) Construction of pCI-AVMVH-CH vector

Using the synthetic gene as a template, the gene fragment in the AVMVH-CH region was amplified by PCR and inserted into the pCI vector (manufactured by Promega) to create the pCI-AVMVH-CH vector.

(2-4) Construction of pCI-AVM-hLG4PE(R409K/S354C/T366W)-Connector-AVMscFv-FLAG Tag Carrier

Using the synthesized gene as a template, the gene fragment of the CH1-hinge-CH2-CH3(R409K/S354C/T366W)-linker region was amplified by PCR. Additionally, using N5LG4PE_AVM as a template, the gene fragment of the linker-AVMscFv-FLAG tag region was amplified by PCR. The gene fragments of the CH1-hinge-CH2-CH3(R409K/S354C/T366W)-linker region and the linker-AVMscFv-FLAG tag region were inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVMscFv to create the pCI-AVM-hLG4PE(R409K/S354C/T366W)-linker-AVMscFv-FLAG tag vector.

(2-5) Construction of pCI-AVM-hLG4PE(R409K)_AVMscFv vector

Using the synthesized gene as a template, the gene fragment of the CH1-hinge-CH2-CH3-connector region was amplified by PCR. Additionally, using N5LG4PE_AVM as a template, the gene fragments of the VH and VL regions of the AVM were amplified by PCR. The gene fragments of the CH1-hinge-CH2-CH3-connector region and the gene fragments of the VH and VL regions of the AVM were inserted into the NheI-BamHI site of pCI-AVM-hLG4PE(R409K)_AVMscFv to create the pCI-AVM-hLG4PE(R409K)_AVMscFv3 and pCI-AVM-hLG4PE(R409K)_AVMscFv5 vectors.

[Example 16] Preparation of various bispecific antibodies

Using the method described in Example 6, the following antibodies were prepared: AVM IgG4PE(R409K)_MOG01Fab antibody, AVM IgG4PE(R409K)_MOG01dscFv2 antibody, AVM IgG4PE(R409K)_MOG01dscFv3 antibody, AVM IgG4PE(R409K)_MOG01dscFv4 antibody, AVM IgG4PE(R409K)_MOG01dscFv5 antibody, AVM IgG4PE(R409K)_MOG01dscFv6 antibody, and AVM IgG4PE(R409K)_MOG01 dscFv6 antibody. The following antibodies were used: dscFv7 antibody, AVM IgG4PE(R409K)_MOG01dscFv8 antibody, AVM IgG4PE(R409K)_MOG01dscFv9 antibody, AVM IgG4PE(R409K)_MOG01dscFv10 antibody, AVM IgG4PE(R409K)_MOG01dscFv11 antibody, AVM IgG4PE(R409K)_AVM Fab antibody, AVM IgG4PE(R409K)_AVMdscFv3 antibody, and AVM IgG4PE(R409K)_AVMdscFv5 antibody.

AVM-MOG01 IgG4PE(R409K) antibody, AVM IgG4PE(R409K)_MOG01sscFv antibody, and AVMIgG4PE(R409K)_AVMsscFv antibody were prepared using the methods described below. The antibody expression plasmid vector was introduced into suspension 293 cells using the Expi293 (trademark) expression system (manufactured by Thermo Fisher Scientific) and cultured to express the antibody in a transient expression system.

Four days after vector introduction, the culture supernatant was recovered and filtered using a 0.22 μm membrane filter (MILLIPORE). Proteins in the culture supernatant were purified using His-tag affinity purification with Ni agarose resin (GE Healthcare Bioscience). 20 mM imidazole-phosphate buffer was used as the washing buffer.

The antibody adsorbed on Ni agarose resin was eluted using 500 mM imidazole-phosphate buffer. Then, the solvent in the eluent was replaced with PBS using ultrafiltration with a VIVASPIN (Sartrius Stealin) and a NAP column (GE Healthcare Bioscience).

The His-tagged protein was purified using a FLAG antibody affinity gel (Sigma-Aldrich). Phosphate buffer was used as the washing solution. The antibody adsorbed on the FLAG antibody affinity gel was eluted with 20 mM sodium citrate and 50 mM NaCl buffer (pH 3.4) and recovered into a tube containing 1 M Tris-HCl buffer (pH 8.0).

Next, the solvent of the eluent was replaced with PBS using ultrafiltration with a VIVASPIN (Sartrius Stealin) and a NAP column (GE Healthcare Bioscience), followed by filtration and sterilization using a 0.22 μm membrane filter (Millex-GV, MillIPORE). The absorbance of the antibody solution at 280 nm was measured to calculate the concentration of the purified antibody.

[Example 17] Evaluation of the binding affinity of various bispecific antibodies to MOG using flow cytometry

The binding of various bispecific antibodies and negative control antibodies obtained in Examples 6 and 16 to MOG was evaluated using fluorescence activated cell sorting (FACS) following the steps described below.

The pEF6_hMOG obtained in Example 5 was introduced into L929 fibroblasts derived from mouse connective tissue using HilyMax (manufactured by Tongren Chemical Co., Ltd.) [American Center for Type Culture Collection (ATCC) No.: CCL-1]. After screening the gene-introduced cells using blastcin (manufactured by Invitrogen) as an antibiotic, cloning was performed using the limiting dilution method. The reactivity of various bispecific antibodies was analyzed using L929 cells expressing hMOG on their cell surface (hereinafter referred to as hMOG/L929) using the methods described below.

hMOG/L929 cells were suspended in staining buffer (SB) containing 0.1% _NaN3 and 1% FBS in PBS and aliquoted into 96-well round-bottom plates (manufactured by BD). After centrifugation (2000 rpm, 4°C, 2 min), the supernatant was removed, and the various MOG01 bispecific antibodies obtained in Examples 6 and 16 were added to the clumps and resuspended. The clumps were then incubated at ice for 30 min. Further centrifugation (2000 rpm, 4°C, 2 min) was performed, the supernatant was removed, and the clumps were washed with SB. 1 μg/mL of RPE-labeled goat anti-human antibody (manufactured by Southern Biotech) was added, and the clumps were incubated at ice for 30 min. After washing with SB, the clumps were resuspended in SB, and the fluorescence intensity of each cell was measured using a flow cytometer FACS CANTO II (manufactured by BD). The results are shown in Figures 12(A)–(C) and Figures 13(A) and (B).

As shown in Figures 12(A)–(C) and 13(A) and (B), various bispecific antibodies were confirmed to bind to MOG. Furthermore, it was shown that, particularly with the AVM IgG4PE(R409K)_MOG01Fab antibody [Figure 10(B), Figure 12(C)], AVM IgG4PE(R409K)_MOG01dscFv3 antibody, AVM IgG4PE(R409K)_MOG01dscFv5 antibody, AVM IgG4PE(R409K)_MOG01dscFv6 antibody, and AVM IgG4PE(R409K)_ High binding activity was observed with the following antibodies: MOG01dscFv7, AVM IgG4PE(R409K)_MOG01dscFv8, AVM IgG4PE(R409K)_MOG01dscFv9, AVM IgG4PE(R409K)_MOG01dscFv10, and AVM IgG4PE(R409K)_MOG01dscFv11 [Figure 11(B), Figure 13(B)].

[Example 18] Evaluation of the binding affinity of various bispecific antibodies to MOG using surface plasmon resonance detection

The binding of the various bispecific antibodies obtained in Examples 6 and 16 to MOG was evaluated using the same method as in Example 8. The results are shown in Tables 10 and 11.

[Table 10]

[Table 11]

As shown in Tables 10 and 11, the dissociation constants (KD values) of the bispecific antibodies for each MOG range from 1.2 × ^10⁻⁸ (M) to 2.0 × ^10⁻⁷ (M), indicating that any one of the antibodies exhibits good affinity.

Furthermore, it was shown that the binding was particularly high with AVM-MOG01 IgG4PE(R409K) antibody [Fig. 10(A)], AVM IgG4PE(R409K)_MOG01Fab antibody [Fig. 10(B)], AVM IgG4PE(R409K)_MOG01dscFv3 antibody, and AVM IgG4PE(R409K)_MOG01dscFv5 antibody [Fig. 11(B)].

[Example 19] Evaluation of the brain migration properties of various bispecific antibodies in mice

The mouse brain migration of various bispecific antibodies and negative control antibodies obtained in Examples 6 and 16 was evaluated using the method of Example 14.

For AVM-MOG01 IgG4PE(R409K) antibody, AVM IgG4PE(R409K)_MOG01sscFv antibody and AVMIgG4PE(R409K)_MOG01Fab antibody, the antibody was administered at 5 mg/kg. The serum antibody concentration and the amount of antibody per unit brain weight in brain tissue after 10 days are shown in Figures 14(A) to 16(B).

As shown in Figures 14(A), 15(A), and 16(A), the serum antibody concentrations of any MOG01 altered antibody were not different from those of the negative control. On the other hand, as shown in Figures 14(B), 15(B), and 16(B), compared with the negative control, the amount of antibody in the brain increased by approximately 8-fold in the case of AVM-MOG01 IgG4PE(R409K) antibody, by approximately 12-fold in the case of AVMIgG4PE(R409K)_MOG01sscFv antibody, and by approximately 30-fold in the case of AVM IgG4PE(R409K)_MOG01Fab antibody.

As shown above, compared with the negative control antibody that does not bind to MOG, various bispecific antibodies that bind to MOG can increase the amount of antibodies in the brain, while the blood half-life remains unchanged.

For AVM IgG4PE(R409K)_MOG01dscFv antibody, AVM IgG4PE(R409K)_MOG01dscFv3 antibody and AVM IgG4PE(R409K)_MOG01dscFv5 antibody, the antibody was administered at 5 mg/kg. The serum antibody concentration and the amount of antibody per unit brain weight in brain tissue after 10 days and 28 days are shown in Figures 17(A) to (D).

As shown in Figures 17(A) and (C), the serum antibody concentration of any bispecific antibody was not different from that of the negative control antibody. However, as shown in Figures 17(B) and (D), the AVM IgG4PE(R409K)_MOG01dscFv antibody, AVM IgG4PE(R409K)_MOG01dscFv3 antibody, and AVM IgG4PE(R409K)_MOG01dscFv5 antibody increased the amount of antibody in the brain by several tens of times over 28 days. Furthermore, as shown in Figure 17(D), the bispecific antibodies with higher antibody levels in the brain after 28 days also showed higher binding to MOG (Table 11), indicating that MOG binding activity is correlated with the amount of antibody in the brain.

[Example 20] Obtaining a novel MOG antibody exhibiting stronger MOG binding than the anti-MOG01 antibody.

(1) Preparation of extracellular domain proteins of soluble human MOG antigen and soluble mouse MOG antigen bound with FLAG-Fc

Plasmid vectors INPEP4_hMOG-FLAG-Fc and INPEP4_mMOG-FLAG-Fc, expressing soluble antigens of human and mouse MOGs with FLAG-Fc appended to the C-terminus, were prepared using the method described in Example 4. The base sequence of hMOG-FLAG-Fc is shown in Serial No. 100, the amino acid sequence deduced from this base sequence is shown in Serial No. 101, the base sequence of mMOG-FLAG-Fc is shown in Serial No. 102, and the amino acid sequence deduced from this base sequence is shown in Serial No. 103. The FLAG-Fc-bound MOG extracellular domain protein was obtained by transient expression and purification using the method described in Example 4.

(2) Production of GST-binding MOG extracellular domain proteins

Plasmid vectors N5_hMOG-GST and N5_mMOG-GST, expressing soluble antigens of human and mouse MOGs and containing GST appended to the C-terminus, were prepared using the method described in Example 4. The base sequence of hMOG-GST is shown in Serial No. 104, the amino acid sequence deduced from this base sequence is shown in Serial No. 105, the base sequence of mMOG-GST is shown in Serial No. 106, and the amino acid sequence deduced from this base sequence is shown in Serial No. 107. The GST-bound MOG extracellular domain protein was obtained by transient expression and purification using the method described in Example 4.

(3) Obtaining anti-MOG antibodies from human antibody-producing mice

hMOG-GST and mMOG-GST were mixed with pertussis vaccine and gum arabic and administered intraperitoneally or intradermally to human antibody-producing mice (Ishida & Lonberg, IBC’s 11th Antibody Engineering, Abstract 2000; Ishida, I. et al., Cloning & Stem Cells 4, 85-96 (2002); Ishida Isao (2002) Experimental Medicine 20, 6, 846-851).

Following the initial immunization, three doses of hMOG-GST and mAMOG-GST were administered. Four days after the final immunization, spleens were dissected and harvested from individuals who received intraperitoneal administration. Red blood cells were removed using a red blood cell removal reagent (SIGMA) and the spleens were frozen in Cellbanker 1 (Zenyasu Kogyo Co., Ltd., Japan). Axillary lymph nodes were dissected and harvested from individuals who received intradermal administration. Red blood cells were removed using a red blood cell removal reagent and the nodes were frozen in Cellbanker 1. RNA was extracted from the spleen cells and axillary lymph node cells using the RNeasy Plus mini-extraction kit (QIAGEN), and cDNA was synthesized using the SMARTer RACE cDNA amplification kit (Clontech). Using the synthesized cDNA, a phage library derived from human antibody-producing mice was constructed using the method described in Example 1.

Using a phage library derived from human antibody-producing mice, anti-human MOG monoclonal antibodies were obtained via phage display. Phage display and cloning ELISA were performed using hMOG-FLAG_Fc and mMOG-FLAG_Fc according to the methods described in Example 1.

Sequence analysis was performed on clones that bound hMOG-FLAG_Fc, mMOG-FLAG_Fc, and hMOG/Expi293F cells to obtain the anti-MOG antibody phagemid vectors pCANTAB_MOG301, pCANTAB_MOG303, pCANTAB_MOG307, pCANTAB_MOG310, pCANTAB_MOG312, pCANTAB_MOG326, pCANTAB_MOG329, pCANTAB_MOG446, pCANTAB_MOG456, and pCANTAB_MOG473.

In the following text, the names of the anti-MOG scFv antibodies displayed by phages expressed using pCANTAB_MOG301, pCANTAB_MOG303, pCANTAB_MOG307, pCANTAB_MOG310, pCANTAB_MOG312, pCANTAB_MOG326, pCANTAB_MOG329, pCANTAB_MOG446, pCANTAB_MOG456, and pCANTAB_MOG473 will be recorded as MOG301 antibody, MOG303 antibody, MOG307 antibody, MOG310 antibody, MOG312 antibody, MOG326 antibody, MOG329 antibody, MOG446 antibody, MOG456 antibody, and MOG473 antibody, respectively.

The base sequences encoding the VH or VL of various anti-MOG antibodies, and the amino acid sequences deduced from these base sequences, are shown in Table 12.

In addition, using phage display with MOG binding as an indicator, clones with similar sequences of 91-93% homology to MOG301 antibody (MOG426, MOG428), 85-95% homology to MOG303 antibody (MOG313, MOG314, MOG315, MOG331, MOG357, MOG476), and 97-99% homology to MOG307 antibody (MOG323, MOG476) were obtained. Clones with sequences highly homologous to the MOG310 antibody (MOG341, MOG354, MOG355), MOG308, MOG316, MOG319, MOG320, MOG338, MOG352, MOG359, MOG478), MOG329 (MOG470), and MOG456 (MOG418) were identified. These similar clones were confirmed to bind to hMOG-FLAG_Fc, mMOG-FLAG_Fc, and hMOG/Expi293F cells, indicating that antibody clones with high amino acid sequence homology to each antibody clone also possess MOG-binding activity.

Table 13 shows the base sequences encoding similar clones VH or VL, and the amino acid sequences deduced from these base sequences. Figures 18 to 22(B) show a comparison of the amino acid sequences of similar clones.

[Example 21] Preparation of anti-MOG scFv-Fc antibody

Using the phage vector pCANTAB_MOG01 as a template, the gene fragment in the scFv region was amplified by PCR. Using the synthetic gene of the heavy chain constant region as a template, the gene fragment in the hinge -CH2-CH3 region was amplified by PCR. The obtained gene fragments were inserted into the N5KG4PE vector (described in International Publication No. 2002/088186) to prepare the N5-MOG01scFv-hG4PE vector.

Using the phage vector pCANTAB_MOG301 as a template, the gene fragment in the scFv region was amplified by PCR. Using the synthetic gene of the heavy chain constant region as a template, the gene fragment in the hinge -CH2-CH3 region was amplified by PCR. The obtained gene fragments were inserted into the pCI vector (manufactured by Promega) to create the pCI-MOG301scFv-hG4PE(R409K) vector.

Antibody expression vectors containing gene fragments inserted into the scFv regions of various anti-MOG antibodies shown in Table 12 were prepared using the same method and named pCI-MOG303scFv-hG4PE(R409K), pCI-MOG307scFv-hG4PE(R409K), pCI-MOG310scFv-hG4PE(R409K), pCI-MOG312scFv-hG4PE(R409K), pCI-MOG326scFv-hG4PE(R409K), pCI-MOG329scFv-hG4PE(R409K), pCI-MOG446scFv-hG4PE(R409K), pCI-MOG456scFv-hG4PE(R409K), and pCI-MOG473scFv-hG4PE(R409K).

The expression vector for the anti-MOG antibody was prepared using the method described in Example 6. The expression vectors for anti-MOG antibodies, pCI-MOG301scFv-hG4PE(R409K), pCI-MOG303scFv-hG4PE(R409K), pCI-MOG307scFv-hG4PE(R409K), pCI-MOG310scFv-hG4PE(R409K), pCI-MOG312scFv-hG4PE(R409K), pCI-MOG326scFv-hG4PE(R409K), pCI-MOG329scFv-hG4PE(R409K), pCI-MOG446scFv-hG4PE(R409K), pCI-MOG456scFv-hG4PE(R409K), and pCI-MOG473scFv-hG4PE(R409K), were developed. G4PE(R409K) was expressed to obtain the following antibodies: MOG301scFv-hG4PE(R409K), MOG303scFv-hG4PE(R409K), MOG307scFv-hG4PE(R409K), MOG310scFv-hG4PE(R409K), MOG312scFv-hG4PE(R409K), MOG326scFv-hG4PE(R409K), MOG329scFv-hG4PE(R409K), MOG446scFv-hG4PE(R409K), MOG456scFv-hG4PE(R409K), and MOG473scFv-hG4PE(R409K).

[Example 22] Evaluation of the binding affinity of anti-MOG antibodies to MOG using flow cytometry

The binding of the anti-MOG antibody obtained in Example 21 to MOG was evaluated using the same method as in Example 7. The results are shown in Figures 23-25.

As shown in Figures 23-25, MOG01scFv-hG4PE, MOG301scFv-hG4PE(R409K), MOG303scFv-hG4PE(R409K), MOG307scFv-hG4PE(R409K), MOG310scFv-hG4PE(R409K), MOG312scFv-hG4PE(R409K), and MOG326s cFv-hG4PE(R409K), MOG329scFv-hG4PE(R409K), MOG446scFv-hG4PE(R409K), MOG456scFv-hG4PE(R409K), and MOG473scFv-hG4PE(R409K) all showed binding activity against hMOG/Expi293F and mMOG/Expi293F cells.

[Example 23] Evaluation of the binding affinity of anti-MOG antibodies to MOG using surface plasmon resonance detection

The binding of MOG01scFv-hG4PE, MOG301scFv-hG4PE(R409K), MOG303scFv-hG4PE(R409K), MOG307scFv-hG4PE(R409K), MOG329scFv-hG4PE(R409K), MOG446scFv-hG4PE(R409K), MOG456scFv-hG4PE(R409K), and MOG473scFv-hG4PE(R409K) to human and mouse MOGs was evaluated using the same method as in Example 8. The analytes used were hMOG-GST and mMOG-GST. The results of the binding evaluation for human MOGs are shown in Table 14, and the results of the binding evaluation for mouse MOGs are shown in Table 15.

[Table 14]

Reactivity to human MOGs

MOG01's ka is outside the measurement range.

[Table 15]

responsiveness to mouse MOG

As shown in Tables 14 and 15, the dissociation constants (KD values) of each anti-MOG antibody for human MOG range from 1.0 × ^10⁻¹⁰ (M) to 3.6 × ^10⁻⁹ (M), and for mouse MOG range from 1.9 × ^10⁻¹⁰ (M) to 6.9 × ^10⁻⁹ (M), indicating that each antibody exhibits good affinity. In the case of MOG01scFv-hG4PE, the binding rate constant ka is outside the measurement range of the equipment, making it impossible to uniquely determine the KD value.

[Example 24] Preparation of enzyme fusion antibody

Enzyme fusion antibodies containing acid sphingomyelinase (ASM) fused to the C-terminus of anti-MOG01 IgG and anti-AVM IgG antibodies were prepared using the methods described below. The expression vector for the anti-MOG01 IgG antibody with ASM fused to the C-terminus was named pCI-MOG01-hLG4PE(R409K)_ASM, and the expression vector for the anti-AVM IgG antibody with ASM fused to the C-terminus was named pCI-AVM-hLG4PE(R409K)_ASM.

Using the synthetic gene of ASM (sequence number 150) as a template, the gene fragment of the adapter-ASM region was amplified by PCR. Additionally, using the synthetic gene as a template, the gene fragment of the CH1-hinge-CH2-CH3(R409K) region was synthesized by PCR. Using N5LG4PE_MOG01 as a template, the gene fragments of the MOG01 light chain region and the MOG01 VH region were amplified by PCR.

The obtained gene fragment was inserted into a pCI vector (manufactured by Promega) to create the pCI-MOG01-hLG4PE(R409K)_ASM vector. Using the synthesized gene as a template, the gene fragment in the CH2-CH3 region was amplified by PCR. The gene fragments in the CH2-CH3 region and the adapter-ASM region were then inserted into the PmlI-BamHI site of the pCI-AVM-hLG4PE(R409K) vector to create pCI-AVM-hLG4PE(R409K)_ASM.

pCI-MOG01-hLG4PE(R409K)_ASM and pCI-AVM-hLG4PE(R409K)_ASM were expressed and purified using the method shown in Example 6. The antibody obtained by expressing pCI-MOG01-hLG4PE(R409K)_ASM was named MOG01 IgG4PE(R409K)-ASM, and the antibody obtained by expressing pCI-AVM-hLG4PE(R409K)_ASM was named AVM IgG4PE(R409K)-ASM.

[Example 25] Activity evaluation of enzyme fusion antibody

The binding affinity of MOG01 IgG4PE(R409K)-ASM to MOG-expressing cells was confirmed using the same method as in Example 23, and the results are shown in Figure 26. Furthermore, the binding affinity to the soluble MOG antigen was confirmed using the same method as in Example 8. The dissociation constant (KD value) of MOG01 IgG4PE(R409K)-ASM was 2.9 × ^10⁻⁹ (M), indicating good affinity.

The binding of the anti-ASM antibody (manufactured by LSBio) to the prepared MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM was confirmed using the ELISA method shown below.

In the ELISA, MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM were immobilized (100 ng/50 μL) on a MAXISORP (NUNC). Sites not bound to MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM were blocked using SuperBlock Blocking Buffer (Thermo). As a negative control, plates immobilized (50 ng/50 μL) with anti-MOG01 IgG antibody and anti-AVM IgG antibody were also prepared. Anti-ASM antibody diluted to 0.2, 1, and 5 μg/mL with PBS-T was added to each well. After reacting for 1 hour at room temperature, each well was washed with PBS-T.

Next, a solution of horseradish peroxidase-labeled anti-mouse immunoglobulin antibody (manufactured by Dako) diluted with PBS-T was added to each well, and the reaction was carried out at room temperature for 1 hour. TMB chromogenic substrate solution (manufactured by Dako) was added, and the mixture was incubated at room temperature. 2M hydrochloric acid was added to each well to terminate the chromogenic reaction, and the absorbance was measured at a wavelength of 450 nm (reference wavelength 570 nm). The results are shown in Figure 27.

Figure 27 shows that the prepared ASM fusion antibody was recognized and bound by the anti-ASM antibody. Furthermore, the sphingomyelinase activities of the prepared MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM were measured using a sphingomyelinase activity assay kit (manufactured by Echelon Biosciences), confirming that the prepared ASM fusion antibody possesses enzymatic activity.

The above results confirm that the antigen-binding activity and enzyme activity of the enzyme fusion antibody formed by fusing the enzyme with the MOG antibody were both maintained.

[Example 26] Evaluation of mouse brain migration of enzyme fusion antibody

The mouse brain migration of the ASM fusion antibody obtained in Example 24 was evaluated using the same method as in Example 14. The ASM fusion antibody was administered at 5 mg/kg, and the serum antibody concentration and the amount of antibody per unit brain weight in brain tissue after 10 days are shown in Figure 28.

As shown in Figure 28, there was no difference in serum antibody concentration between MOG01 IgG4PE(R409K)-ASM and AVM IgG4PE(R409K)-ASM. On the other hand, it was shown that in the case of MOG01 IgG4PE(R409K)-ASM, the amount of antibody in the brain increased by approximately 58 times compared with AVM IgG4PE(R409K)-ASM.

The present invention has been described in detail using specific methods, but it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the intent and scope of the invention. It should be noted that this application is based on Japanese Patent Application No. 2016-251106, filed on December 26, 2016, the entire contents of which are incorporated herein by reference.

Sequence List Free Text

Description of Serial Number 3 - Artificial Sequence: The amino acid sequence of VH of MOG01 without the signal sequence.

Description of Serial Number 4 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG01

Description of Serial Number 5 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG01

Description of Serial Number 6 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG01

Description of Serial Number 9 - Artificial Sequence: The amino acid sequence of VL of MOG01 without the signal sequence.

Description of Serial Number 10 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG01.

Description of Serial Number 11 - Artificial Sequence: Amino acid sequence of LCDR2 of MOG01.

Description of Serial Number 12 - Artificial Sequence: Amino acid sequence of LCDR3 of MOG01.

Description of Serial Number 15 - Artificial Sequence: The amino acid sequence of VH of MOG09 without the signal sequence.

Description of Serial Number 16 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG09.

Description of Serial Number 17 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG09.

Description of Serial Number 18 - Artificial Sequence: Amino acid sequence of HCDR3 of MOG09.

Description of Serial Number 21 - Artificial Sequence: The amino acid sequence of MOG09 VL without the signal sequence.

Description of Serial Number 22 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG09.

Description of Serial Number 23 - Artificial Sequence: Amino acid sequence of LCDR2 of MOG09.

Description of Serial Number 24 - Artificial Sequence: Amino acid sequence of LCDR3 of MOG09.

Description of Serial Number 27 - Artificial Sequence: The amino acid sequence of VH of MOG14 without the signal sequence.

Description of Serial Number 28 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG14

Description of Serial Number 29 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG14.

Description of Serial Number 30 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG14.

Description of Serial Number 33 - Artificial Sequence: The amino acid sequence of VL of MOG14 without the signal sequence.

Description of Serial Number 34 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG14.

Description of Serial Number 35 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG14.

Description of Serial Number 36 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG14.

Description of Serial Number 37 - Artificial Sequence: VHH base sequence of iMOG_3Rim1_S32 containing the signal sequence.

Description of Serial Number 38 - Artificial Sequence: Amino acid sequence of the synthetic construct.

Description of Serial Number 39 - Artificial Sequence: The amino acid sequence of VHH in iMOG_3Rim1_S32 that does not contain the signal sequence.

Description of Serial Number 40 - Artificial Sequence: Amino acid sequence of CDR1 in iMOG_3Rim1_S32

Description of Serial Number 41 - Artificial Sequence: Amino acid sequence of CDR2 of iMOG_3Rim1_S32

Description of Serial Number 42 - Artificial Sequence: Amino acid sequence of CDR3 in iMOG_3Rim1_S32

Description of Serial Number 43 - Artificial Sequence: Base sequence of Primer 1

Description of sequence number 44 - artificial sequence: base sequence of primer 2

Description of sequence number 45 - artificial sequence: Base sequence of primer 3

Description of sequence number 46 - artificial sequence: Base sequence of primer 4

Description of Serial Number 47 - Artificial Sequence: Base sequence of Primer 5

Description of sequence number 48 - artificial sequence: Base sequence of primer 6

Description of sequence number 49 - artificial sequence: Base sequence of primer 7

Description of sequence number 50 - artificial sequence: Base sequence of primer 8

Description of sequence number 51 - artificial sequence: Base sequence of primer 9

Description of Serial Number 52 - Artificial Sequence: Base sequence of primer 10

Description of Serial Number 53 - Artificial Sequence: Base sequence of primer 11

Description of sequence number 54 - artificial sequence: Base sequence of primer 12

Description of sequence number 55 - artificial sequence: Base sequence of primer 13

Description of sequence number 56 - artificial sequence: Base sequence of primer 14

Description of sequence number 57 - artificial sequence: Base sequence of primer 15

Description of sequence number 58 - artificial sequence: Base sequence of primer 16

Description of sequence number 59 - artificial sequence: Base sequence of primer 17

Description of sequence number 60 - artificial sequence: Base sequence of primer 18

Description of sequence number 61 - artificial sequence: Base sequence of primer 19

Description of sequence number 62 - artificial sequence: Base sequence of primer 20

Description of sequence number 63 - artificial sequence: Base sequence of primer 21

Description of sequence number 64 - artificial sequence: base sequence of primer 22

Description of sequence number 65 - artificial sequence: Base sequence of primer 23

Description of sequence number 66 - artificial sequence: Base sequence of primer 24

Description of sequence number 69 - artificial sequence: base sequence of rMOG-FLAG-Fc

Description of Serial Number 70 - Artificial Sequence: Amino acid sequence of the synthetic construct.

Description of Serial Number 71 - Artificial Sequence: Base sequence of rMOG-GST

Description of Serial Number 72 - Artificial Sequence: Amino acid sequence of the synthetic construct

Description of sequence number 79 - artificial sequence: Base sequence of primer 25

Description of sequence number 80 - artificial sequence: Base sequence of primer 26

Description of sequence number 81 - artificial sequence: Base sequence of primer 27

Description of sequence number 82 - artificial sequence: Base sequence of primer 28

Description of sequence number 83 - artificial sequence: Base sequence of primer 29

Description of sequence number 84 - artificial sequence: Base sequence of primer 30

Description of sequence number 85 - artificial sequence: Base sequence of primer 31

Description of sequence number 86 - artificial sequence: Base sequence of primer 32

Description of sequence number 87 - artificial sequence: Base sequence of primer 33

Description of sequence number 88 - artificial sequence: Base sequence of primer 34

Description of sequence number 89 - artificial sequence: Base sequence of primer 35

Description of sequence number 90 - artificial sequence: Base sequence of primer 36

Description of sequence number 91 - artificial sequence: Base sequence of primer 37

Description of sequence number 92 - artificial sequence: Base sequence of primer 38

Description of sequence number 93 - artificial sequence: Base sequence of primer 39

Description of sequence number 94 - artificial sequence: Base sequence of primer 40

Description of sequence number 95 - artificial sequence: Base sequence of primer 41

Description of sequence number 96 - artificial sequence: Base sequence of primer 42

Description of sequence number 97 - artificial sequence: Base sequence of primer 43

Description of Serial Number 98 - Artificial Sequence: Base sequence of hHER2-GST

Description of Serial Number 99 - Artificial Sequence: Amino acid sequence of the synthetic construct.

Description of sequence number 100 - artificial sequence: base sequence of hMOG-FLAG-Fc (including signal sequence).

Description of sequence number 101 - artificial sequence: amino acid sequence of hMOG-FLAG-Fc (including signal sequence).

Description of sequence number 102 - artificial sequence: base sequence of mMOG-FLAG-Fc (including signal sequence).

Description of sequence number 103 - artificial sequence: amino acid sequence of mMOG-FLAG-Fc (including signal sequence).

Description of sequence number 104 - artificial sequence: base sequence of hMOG-GST (including signal sequence).

Description of Serial Number 105 - Artificial Sequence: Amino acid sequence of hMOG-GST (including signal sequence)

Description of sequence number 106 - artificial sequence: base sequence of mMOG-GST (including signal sequence).

Description of sequence number 107 - artificial sequence: amino acid sequence of mMOG-GST (including signal sequence).

Description of Serial Number 108 - Artificial Sequence: The base sequence of the antibody sequence with the pCI-AVM-hLG4PE(R409K/S354C/T366W)-FLAG tag (excluding the signal sequence).

Description of Serial Number 109 - Artificial Sequence: Amino acid sequence of the antibody sequence with the pCI-AVM-hLG4PE(R409K/S354C/T366W)-FLAG tag (excluding the signal sequence).

Description of Serial Number 110 - Artificial Sequence: pCI-MOG01-hLG4PE(R409K/Y349C/T366S/L368A/Y407V) - Base sequence of the His-tagged antibody sequence (excluding the signal sequence).

Description of Serial Number 111 - Artificial Sequence: pCI-MOG01-hLG4PE(R409K/Y349C/T366S/L368A/Y407V) - Amino acid sequence of the His-tagged antibody sequence (excluding the signal sequence).

Description of sequence number 112 - artificial sequence: Base sequence of the antibody sequence of pCI-AVM-hLG4PE(R409K)-linker-MOG01VL-CL (excluding signal sequence).

Description of Serial Number 113 - Artificial Sequence: Amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)-Linker-MOG01VL-CL (excluding signal sequence).

Description of sequence number 114 - artificial sequence: Base sequence of the antibody sequence pCI-MOG01VH-CH (excluding signal sequence).

Description of sequence number 115 - artificial sequence: amino acid sequence of the antibody sequence pCI-MOG01VH-CH (excluding signal sequence).

Description of sequence number 116 - artificial sequence: pCI-AVM-hLG4PE(R409K/S354C/T366W) - adapter - MOG01scFv - the base sequence of the FLAG-tagged antibody sequence (excluding the signal sequence).

Description of Serial Number 117 - Artificial Sequence: Amino acid sequence of the antibody sequence with the pCI-AVM-hLG4PE(R409K/S354C/T366W)-Connector-MOG01scFv-FLAG tag (excluding signal sequence).

Description of sequence number 118 - artificial sequence: pCI-AVM-hLG4PE(R409K/Y349C/T366S/L368A/Y407V) - the base sequence of the His-tagged antibody sequence (excluding the signal sequence).

Description of Serial Number 119 - Artificial Sequence: pCI-AVM-hLG4PE(R409K/Y349C/T366S/L368A/Y407V) - Amino acid sequence of the His-tagged antibody sequence (excluding the signal sequence).

Description of sequence number 120 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv2 (excluding signal sequence).

Description of sequence number 121 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv2 (excluding signal sequence).

Description of sequence number 122 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv3 (excluding signal sequence).

Description of sequence number 123 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv3 (excluding signal sequence).

Description of sequence number 124 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv4 (excluding signal sequence).

Description of sequence number 125 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv4 (excluding signal sequence).

Description of sequence number 126 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv5 (excluding signal sequence).

Description of sequence number 127 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv5 (excluding signal sequence).

Description of sequence number 128 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv6 (excluding signal sequence).

Description of sequence number 129 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv6 (excluding signal sequence).

Description of sequence number 130 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv7 (excluding signal sequence).

Description of sequence number 131 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv7 (excluding signal sequence).

Description of sequence number 132 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv8 (excluding signal sequence).

Description of sequence number 133 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv8 (excluding signal sequence).

Description of sequence number 134 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv9 (excluding signal sequence).

Description of sequence number 135 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv9 (excluding signal sequence).

Description of sequence number 136 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv10 (excluding signal sequence).

Description of sequence number 137 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv10 (excluding signal sequence).

Description of sequence number 138 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv11 (excluding signal sequence).

Description of sequence number 139 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_MOG01scFv11 (excluding signal sequence).

Description of sequence number 140 - artificial sequence: Base sequence of the antibody sequence of pCI-AVM-hLG4PE(R409K)-linker-AVMVL-CL (excluding signal sequence).

Description of sequence number 141 - artificial sequence: amino acid sequence of the antibody sequence of pCI-AVM-hLG4PE(R409K)-linker-AVMVL-CL (excluding signal sequence).

Description of sequence number 142 - artificial sequence: Base sequence of the antibody sequence pCI-AVMVH-CH (excluding the signal sequence).

Description of sequence number 143 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVMVH-CH (excluding signal sequence).

Sequence number 144 - Description of the artificial sequence: pCI-AVM-hLG4PE(R409K/S354C/T366W) - adapter - AVMscFv - the base sequence of the FLAG-tagged antibody sequence (excluding the signal sequence).

Description of sequence number 145 - artificial sequence: amino acid sequence of antibody sequence with linker - AVMscFv - FLAG tag (excluding signal sequence) pCI-AVM-hLG4PE(R409K/S354C/T366W) - linker - FLAG tag (excluding signal sequence).

Description of sequence number 146 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_AVMscFv3 (excluding signal sequence).

Description of sequence number 147 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_AVMscFv3 (excluding signal sequence).

Description of sequence number 148 - artificial sequence: Base sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_AVMscFv5 (excluding signal sequence).

Description of sequence number 149 - artificial sequence: amino acid sequence of the antibody sequence pCI-AVM-hLG4PE(R409K)_AVMscFv5 (excluding signal sequence).

Description of Serial Number 150 - Artificial Sequence: Base sequence of acid sphingomyelinase (ASM).

Description of Serial Number 151 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG301.

Description of Serial Number 152 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG301.

Description of Serial Number 153 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG301.

Description of Serial Number 154 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG301.

Description of Serial Number 155 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG301.

Description of Serial Number 156 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG301.

Description of Serial Number 157 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG301.

Description of Serial Number 158 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG301.

Description of Serial Number 159 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG301.

Description of Serial Number 160 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG301.

Description of Serial Number 161 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG303.

Description of Serial Number 162 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG303.

Description of Serial Number 163 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG303.

Description of Serial Number 164 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG303.

Description of sequence number 165 - artificial sequence: amino acid sequence of HCDR3 in MOG303.

Description of Serial Number 166 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG303.

Description of Serial Number 167 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG303.

Description of Serial Number 168 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG303.

Description of Serial Number 169 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG303.

Description of Serial Number 170 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG303.

Description of Serial Number 171 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG307.

Description of Serial Number 172 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG307.

Description of Serial Number 173 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG307.

Description of sequence number 174 - artificial sequence: amino acid sequence of HCDR2 in MOG307.

Description of Serial Number 175 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG307.

Description of Serial Number 176 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG307.

Description of Serial Number 177 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG307.

Description of Serial Number 178 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG307.

Description of Serial Number 179 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG307.

Description of Serial Number 180 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG307.

Description of Serial Number 181 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG310.

Description of Serial Number 182 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG310.

Description of Serial Number 183 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG310.

Description of Serial Number 184 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG310.

Description of Serial Number 185 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG310.

Description of Serial Number 186 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG310.

Description of Serial Number 187 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG310.

Description of Serial Number 188 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG310.

Description of Serial Number 189 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG310.

Description of Serial Number 190 - Artificial Sequence: Amino acid sequence of LCDR3 of MOG310.

Description of Serial Number 191 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG312.

Description of sequence number 192 - artificial sequence: amino acid sequence of VH (excluding signal sequence) of MOG312.

Description of sequence number 193 - artificial sequence: amino acid sequence of HCDR1 in MOG312.

Description of sequence number 194 - artificial sequence: amino acid sequence of HCDR2 in MOG312.

Description of sequence number 195 - artificial sequence: amino acid sequence of HCDR3 in MOG312.

Description of sequence number 196 - artificial sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG312.

Description of sequence number 197 - artificial sequence: amino acid sequence of VL (excluding signal sequence) of MOG312.

Description of sequence number 198 - artificial sequence: amino acid sequence of LCDR1 in MOG312.

Description of sequence number 199 - artificial sequence: amino acid sequence of LCDR2 of MOG312.

Description of Serial Number 200 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG312.

Description of Serial Number 201 - Artificial Sequence: The base sequence encoding the VH (excluding the signal sequence) of MOG326.

Description of Serial Number 202 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG326.

Description of Serial Number 203 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG326.

Description of Serial Number 204 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG326.

Description of Serial Number 205 - Artificial Sequence: The amino acid sequence of HCDR3 in MOG326.

Description of Serial Number 206 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG326.

Description of Serial Number 207 - Artificial Sequence: The amino acid sequence of the VL (excluding the signal sequence) of MOG326.

Description of sequence number 208 - artificial sequence: amino acid sequence of LCDR1 of MOG326.

Description of sequence number 209 - artificial sequence: amino acid sequence of LCDR2 of MOG326.

Description of Serial Number 210 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG326.

Description of Serial Number 211 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG329.

Description of Serial Number 212 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG329.

Description of Serial Number 213 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG329.

Description of Serial Number 214 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG329.

Description of Serial Number 215 - Artificial Sequence: The amino acid sequence of HCDR3 in MOG329.

Description of Serial Number 216 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG329.

Description of Serial Number 217 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG329.

Description of Serial Number 218 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG329.

Description of Serial Number 219 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG329.

Description of Serial Number 220 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG329.

Description of Serial Number 221 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG446.

Description of Serial Number 222 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG446.

Description of Serial Number 223 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG446.

Description of Serial Number 224 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG446.

Description of Serial Number 225 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG446.

Description of Serial Number 226 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG446.

Description of Serial Number 227 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG446.

Description of Serial Number 228 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG446.

Description of Serial Number 229 - Artificial Sequence: Amino acid sequence of LCDR2 of MOG446.

Description of Serial Number 230 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG446.

Description of Serial Number 231 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG456.

Description of Serial Number 232 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG456.

Description of sequence number 233 - artificial sequence: amino acid sequence of HCDR1 in MOG456.

Description of sequence number 234 - artificial sequence: amino acid sequence of HCDR2 in MOG456.

Description of sequence number 235 - artificial sequence: amino acid sequence of HCDR3 of MOG456.

Description of Serial Number 236 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG456.

Description of Serial Number 237 - Artificial Sequence: Amino acid sequence of VL (excluding signal sequence) of MOG456.

Description of Serial Number 238 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG456.

Description of Serial Number 239 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG456.

Description of sequence number 240 - artificial sequence: amino acid sequence of LCDR3 of MOG456.

Description of Serial Number 241 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG473.

Description of Serial Number 242 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG473.

Description of Serial Number 243 - Artificial Sequence: Amino acid sequence of HCDR1 in MOG473.

Description of Serial Number 244 - Artificial Sequence: Amino acid sequence of HCDR2 in MOG473.

Description of Serial Number 245 - Artificial Sequence: Amino acid sequence of HCDR3 in MOG473.

Description of Serial Number 246 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG473.

Description of Serial Number 247 - Artificial Sequence: Amino acid sequence of VL (excluding signal sequence) of MOG473.

Description of Serial Number 248 - Artificial Sequence: Amino acid sequence of LCDR1 in MOG473.

Description of Serial Number 249 - Artificial Sequence: Amino acid sequence of LCDR2 in MOG473.

Description of Serial Number 250 - Artificial Sequence: Amino acid sequence of LCDR3 in MOG473.

Description of Serial Number 251 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG426.

Description of Serial Number 252 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG426.

Description of Serial Number 253 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG426.

Description of Serial Number 254 - Artificial Sequence: Amino acid sequence of VL (excluding signal sequence) of MOG426.

Description of Serial Number 255 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG428.

Description of Serial Number 256 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG428.

Description of Serial Number 257 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG428.

Description of Serial Number 258 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG428.

Description of Serial Number 259 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG313.

Description of Serial Number 260 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG313.

Description of Serial Number 261 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG313.

Description of Serial Number 262 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG313.

Description of Serial Number 263 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG314.

Description of Serial Number 264 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG314.

Description of Serial Number 265 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG314.

Description of Serial Number 266 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG314.

Description of Serial Number 267 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG315.

Description of Serial Number 268 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG315.

Description of Serial Number 269 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG315.

Description of Serial Number 270 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG315.

Description of Serial Number 271 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG331.

Description of Serial Number 272 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG331.

Description of Serial Number 273 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG331.

Description of Serial Number 274 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG331.

Description of Serial Number 275 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG357.

Description of Serial Number 276 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG357.

Description of Serial Number 277 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG357.

Description of Serial Number 278 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG357.

Description of Serial Number 279 - Artificial Sequence: The base sequence encoding the VH (excluding the signal sequence) of MOG476.

Description of Serial Number 280 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG476.

Description of Serial Number 281 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG476.

Description of Serial Number 282 - Artificial Sequence: Amino acid sequence of VL (excluding signal sequence) of MOG476.

Description of Serial Number 283 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG323.

Description of Serial Number 284 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG323.

Description of Serial Number 285 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG323.

Description of Serial Number 286 - Artificial Sequence: Amino acid sequence of VL (excluding signal sequence) of MOG323.

Description of Serial Number 287 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG341.

Description of Serial Number 288 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG341.

Description of Serial Number 289 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG341.

Description of Serial Number 290 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG341.

Description of Serial Number 291 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG354.

Description of Serial Number 292 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG354.

Description of Serial Number 293 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG354.

Description of Serial Number 294 - Artificial Sequence: Amino acid sequence of VL (excluding signal sequence) of MOG354.

Description of Serial Number 295 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG355.

Description of sequence number 296 - artificial sequence: amino acid sequence of VH (excluding signal sequence) of MOG355.

Description of Serial Number 297 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG355.

Description of Serial Number 298 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG355.

Description of Serial Number 299 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG308.

Description of Serial Number 300 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG308.

Description of Serial Number 301 - Artificial Sequence: The base sequence encoding the VL (excluding the signal sequence) of MOG308.

Description of Serial Number 302 - Artificial Sequence: The amino acid sequence of the VL (excluding the signal sequence) of MOG308.

Description of Serial Number 303 - Artificial Sequence: The base sequence encoding the VH (excluding the signal sequence) of MOG316.

Description of Serial Number 304 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG316.

Description of Serial Number 305 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG316.

Description of Serial Number 306 - Artificial Sequence: The amino acid sequence of the VL (excluding the signal sequence) of MOG316.

Description of Serial Number 307 - Artificial Sequence: The base sequence encoding the VH (excluding the signal sequence) of MOG319.

Description of Serial Number 308 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG319.

Description of Serial Number 309 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG319.

Description of Serial Number 310 - Artificial Sequence: The amino acid sequence of the VL (excluding the signal sequence) of MOG319.

Description of Serial Number 311 - Artificial Sequence: The base sequence encoding the VH (excluding the signal sequence) of MOG320.

Description of Serial Number 312 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG320.

Description of Serial Number 313 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG320.

Description of Serial Number 314 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG320.

Description of Serial Number 315 - Artificial Sequence: The base sequence encoding the VH (excluding the signal sequence) of MOG338.

Description of Serial Number 316 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG338.

Description of Serial Number 317 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG338.

Description of Serial Number 318 - Artificial Sequence: The amino acid sequence of the VL (excluding the signal sequence) of MOG338.

Description of Serial Number 319 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG352.

Description of Serial Number 320 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG352.

Description of Serial Number 321 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG352.

Description of Serial Number 322 - Artificial Sequence: Amino acid sequence of the VL (excluding signal sequence) of MOG352.

Description of Serial Number 323 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG359.

Description of Serial Number 324 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG359.

Description of Serial Number 325 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG359.

Description of Serial Number 326 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG359.

Description of Serial Number 327 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG478.

Description of Serial Number 328 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG478.

Description of Serial Number 329 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG478.

Description of Serial Number 330 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG478.

Description of Serial Number 331 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG470.

Description of Serial Number 332 - Artificial Sequence: Amino acid sequence of VH (excluding signal sequence) of MOG470.

Description of Serial Number 333 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG470.

Description of Serial Number 334 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG470.

Description of Serial Number 335 - Artificial Sequence: Base sequence encoding the VH (excluding the signal sequence) of MOG418.

Description of Serial Number 336 - Artificial Sequence: The amino acid sequence of VH (excluding the signal sequence) of MOG418.

Description of Serial Number 337 - Artificial Sequence: Base sequence encoding the VL (excluding the signal sequence) of MOG418.

Description of Serial Number 338 - Artificial Sequence: Amino acid sequence of the VL (excluding the signal sequence) of MOG418.

sequence list

<110> Kyowa Hakko Kirin Co., Ltd.

Kagoshima University (National University Corporation)

<120> Antibody binding to Myelin-oligodendrocyte glycoprotein

<130> W524019

<140> JP2016-251106

<141> 2016-12-26

<160> 338

<170> PatentIn version 3.5

<210> 1

<211> 438

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(438)

<400> 1

atg aac ctc ggg ctc agt ttg att ttc ctt gcc ctc att tta aaa ggt 48

Met Asn Leu Gly Leu Ser Leu Ile Phe Leu Ala Leu Ile Leu Lys Gly

1 5 10 15

gtc cag tgt cag gta cag ctg cag cag tca ggc gca gga tta ttg aag 96

Val Gln Cys Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys

20 25 30

cct tcg gag acc ctt tcc ctc acc tgc gct gtg tct ggt ggg tcc ttc 144

Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe

35 40 45

agt ggt tac tac tgg acc tgg atc cgc cag cgc cca ggg aag ggg ctg 192

Ser Gly Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu

50 55 60

gag tgg att gga gaa atc aat cat cgt gga agc acc gat tac aac ccg 240

Glu Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro

65 70 75 80

tcc ctc aag agt cga gtc acc atg tca ata gac acg tcc aag agc cag 288

Ser Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln

85 90 95

ttc tcc ctg aat ttg aaa tct gtg acc gcc gcg gac acg gct gtg tat 336

Phe Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Ala Asp Thr Ala Val Tyr

100 105 110

tac tgt gcg aga gcc gcc tgg ggg tct tgt tat gat ggg acc tgc tac 384

Tyr Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr

115 120 125

ccc gct gaa tac ttc caa tac tgg ggc cag gga acc ctg gtc acc gtc 432

Pro Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

130 135 140

tcc tca 438

Ser Ser

145

<210> 2

<211> 146

<212> PRT

<213> Homo sapiens

<400> 2

Met Asn Leu Gly Leu Ser Leu Ile Phe Leu Ala Leu Ile Leu Lys Gly

1 5 10 15

Val Gln Cys Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys

20 25 30

Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe

35 40 45

Ser Gly Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu

50 55 60

Glu Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro

65 70 75 80

Ser Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln

85 90 95

Phe Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Ala Asp Thr Ala Val Tyr

100 105 110

Tyr Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr

115 120 125

Pro Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

130 135 140

Ser Ser

145

<210> 3

<211> 127

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VH

of MOG01 excluding signal sequence

<400> 3

Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu

65 70 75 80

Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu

100 105 110

Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 4

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR1 of MOG01

<400> 4

Gly Tyr Tyr Trp Thr

1 5

<210> 5

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR2 of MOG01

<400> 5

Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 6

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR3 of MOG01

<400> 6

Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu Tyr

1 5 10 15

Phe Gln Tyr

<210> 7

<211> 390

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(390)

<400> 7

atg aag ttg cct gtt agg ctg ttg gtg ctg atg ttc tgg att cct gct 48

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala

1 5 10 15

tcc agc agt cag tct gcc ctg act cag cct gcc tcc gtg tct ggg tct 96

Ser Ser Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser

20 25 30

cct gga cag tcg atc acc atc tcc tgc act gga acc agc cgt gac gtt 144

Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val

35 40 45

ggt ggt tat aac tat gtc tcc tgg tac caa caa cac cca ggc aaa gcc 192

Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala

50 55 60

ccc aaa ctc atg att tat gat gtc aat aat cgg ccc tca ggg gtt tct 240

Pro Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser

65 70 75 80

aat cgg ttc tct ggc tcc aag tct ggc aac acg gcc tcc ctg acc atc 288

Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile

85 90 95

tct ggg ctc cag gct gag gac gag gct gat tat ttc tgc agc tca tat 336

Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr

100 105 110

aca agc agt agc acc cct gtg gta ttc ggc ggt ggg acc aag ctg acc 384

Thr Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr

115 120 125

GTC CTA 390

Val Leu

130

<210> 8

<211> 130

<212> PRT

<213> Homo sapiens

<400> 8

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala

1 5 10 15

Ser Ser Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser

20 25 30

Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val

35 40 45

Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala

50 55 60

Pro Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser

65 70 75 80

Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile

85 90 95

Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr

100 105 110

Thr Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr

115 120 125

Val Leu

130

<210> 9

<211> 111

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VL

of MOG01 excluding signal sequence

<400> 9

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser

85 90 95

Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 10

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR1 of MOG01

<400> 10

Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn Tyr Val Ser

1 5 10

<210> 11

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR2 of MOG01

<400> 11

Asp Val Asn Asn Arg Pro Ser

1 5

<210> 12

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR3 of MOG01

<400> 12

Ser Ser Tyr Thr Pro Ser Ser Ser Thr Pro Val Val

1 5 10

<210> 13

<211> 423

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(423)

<400> 13

atg aac ctc ggg ctc agt ttg att ttc ctt gcc ctc att tta aaa ggt 48

Met Asn Leu Gly Leu Ser Leu Ile Phe Leu Ala Leu Ile Leu Lys Gly

1 5 10 15

gtc cag tgt cag gtg cag ctg cag gag tcg ggc cca gga ctg gtg aag 96

Val Gln Cys Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys

20 25 30

tct tcg gag acc ctg tcc ctc acc tgc gct gtc tct ggt cac tcc atc 144

Ser Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly His Ser Ile

35 40 45

agc agt gct tac tac tgg ggc tgg atc cgg cag ccc cca ggg aag ggg 192

Ser Ser Ala Tyr Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly

50 55 60

ctg gag tgg ctt ggg agt att tat cat agt ggg aac acc tac tac aac 240

Leu Glu Trp Leu Gly Ser Ile Tyr His Ser Gly Asn Thr Tyr Tyr Asn

65 70 75 80

ccg tcc ctc aag agt cga gtc acc ata tca gta gac acg tcc aag aac 288

Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn

85 90 95

cag ttc tcc ctg agg ctg acc tct gtg acc gcc gca gac acg gcc gtg 336

Gln Phe Ser Leu Arg Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val

100 105 110

tat tac tgt gcg aga ggg cgt gga tat agt ggc tac gat agc ggt atg 384

Tyr Tyr Cys Ala Arg Gly Arg Gly Tyr Ser Gly Tyr Asp Ser Gly Met

115 120 125

gac gtc tgg ggc caa ggg acc acg gtc acc gtc tcc tca 423

Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

130 135 140

<210> 14

<211> 141

<212> PRT

<213> Homo sapiens

<400> 14

Met Asn Leu Gly Leu Ser Leu Ile Phe Leu Ala Leu Ile Leu Lys Gly

1 5 10 15

Val Gln Cys Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys

20 25 30

Ser Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly His Ser Ile

35 40 45

Ser Ser Ala Tyr Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly

50 55 60

Leu Glu Trp Leu Gly Ser Ile Tyr His Ser Gly Asn Thr Tyr Tyr Asn

65 70 75 80

Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn

85 90 95

Gln Phe Ser Leu Arg Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val

100 105 110

Tyr Tyr Cys Ala Arg Gly Arg Gly Tyr Ser Gly Tyr Asp Ser Gly Met

115 120 125

Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

130 135 140

<210> 15

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VH

of MOG09 excluding signal sequence

<400> 15

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Ser Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly His Ser Ile Ser Ser Ala

20 25 30

Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

35 40 45

Leu Gly Ser Ile Tyr His Ser Gly Asn Thr Tyr Tyr Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Arg Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Cys

85 90 95

Ala Arg Gly Arg Gly Tyr Ser Gly Tyr Asp Ser Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 16

<211> 6

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR1 of MOG09

<400> 16

Ser Ala Tyr Tyr Trp Gly

1 5

<210> 17

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR2 of MOG09

<400> 17

Ser Ile Tyr His Ser Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 18

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR3 of MOG09

<400> 18

Gly Arg Gly Tyr Ser Gly Tyr Asp Ser Gly Met Asp Val

1 5 10

<210> 19

<211> 387

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(387)

<400> 19

atg aag ttg cct gtt agg ctg ttg gtg ctg atg ttc tgg att cct gct 48

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala

1 5 10 15

tcc agc agt tcc tat gtg ctg act cag cca ccc tca gcg tct ggg acc 96

Ser Ser Ser Ser Tyr Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr

20 25 30

ccc ggg cag agg gtc acc atc tct tgt tct gga acc agc tcc aac atc 144

Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Thr Ser Ser Asn Ile

35 40 45

gga atc aat agt gta aac tgg tat caa cag ctc cca gga atg gcc ccc 192

Gly Ile Asn Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Met Ala Pro

50 55 60

aaa ctc gtc atc tac agt agg gat cag cgg ccc tca ggg gtc cct gac 240

Lys Leu Val Ile Tyr Ser Arg Asp Gln Arg Pro Ser Gly Val Pro Asp

65 70 75 80

cga ttc tct ggc tcc cag tct ggc acc tca gcc tcc ctg gcc atc aat 288

Arg Phe Ser Gly Ser Gln Ser Gly Thr Ser Ala Ser Leu Ala Ile Asn

85 90 95

ggc ctc cag tct gag gat gag gct gat tat tgg tgt tca aca tgg gat 336

Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Trp Cys Ser Thr Trp Asp

100 105 110

gac agc ctg aat ggt tgg gtg ttc ggc gga ggg acc aag ctg acc gtc 384

Asp Ser Leu Asn Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val

115 120 125

cta 387

Leu

<210> 20

<211> 129

<212> PRT

<213> Homo sapiens

<400> 20

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala

1 5 10 15

Ser Ser Ser Ser Tyr Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr

20 25 30

Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Thr Ser Ser Asn Ile

35 40 45

Gly Ile Asn Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Met Ala Pro

50 55 60

Lys Leu Val Ile Tyr Ser Arg Asp Gln Arg Pro Ser Gly Val Pro Asp

65 70 75 80

Arg Phe Ser Gly Ser Gln Ser Gly Thr Ser Ala Ser Leu Ala Ile Asn

85 90 95

Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Trp Cys Ser Thr Trp Asp

100 105 110

Asp Ser Leu Asn Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val

115 120 125

Leu

<210> 21

<211> 110

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VL

of MOG09 excluding signal sequence

<400> 21

Ser Tyr Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Thr Ser Ser Asn Ile Gly Ile Asn

20 25 30

Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Met Ala Pro Lys Leu Val

35 40 45

Ile Tyr Ser Arg Asp Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gln Ser Gly Thr Ser Ala Ser Leu Ala Ile Asn Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Trp Cys Ser Thr Trp Asp Asp Ser Leu

85 90 95

Asn Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 22

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR1 of MOG09

<400> 22

Ser Gly Thr Ser Ser Asn Ile Gly Ile Asn Ser Val Asn

1 5 10

<210> 23

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR2 of MOG09

<400> 23

Ser Arg Asp Gln Arg Pro Ser

1 5

<210> 24

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR3 of MOG09

<400> 24

Ser Thr Trp Asp Asp Ser Leu Asn Gly Trp Val

1 5 10

<210> 25

<211> 414

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(414)

<400> 25

atg aac ctc ggg ctc agt ttg att ttc ctt gcc ctc att tta aaa ggt 48

Met Asn Leu Gly Leu Ser Leu Ile Phe Leu Ala Leu Ile Leu Lys Gly

1 5 10 15

gtc cag tgt cag gtg cag ctg gtg caa tct ggg gct gag gtg aag aag 96

Val Gln Cys Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys

20 25 30

cct ggg gcc tca gtg aag gtc tcc tgc cag gct tct gga tac acg ttc 144

Pro Gly Ala Ser Val Lys Val Ser Cys Gln Ala Ser Gly Tyr Thr Phe

35 40 45

acc ggc gac tat att cac tgg gtg cga cag gcc cct gga caa ggg ctg 192

Thr Gly Asp Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu

50 55 60

gaa tac ttg gga tgg atc aac cct gac agg ggt ttc aca tac tat aca 240

Glu Tyr Leu Gly Trp Ile Asn Pro Asp Arg Gly Phe Thr Tyr Tyr Thr

65 70 75 80

cag aag ttt cag ggc agg gtc acc atg acc cgg gac acg tcc agc aac 288

Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Asn

85 90 95

aca gcc tac atg gag ctg agc agc ctg aga tct gac gac acg gcc atg 336

Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Met

100 105 110

tat tac tgt acg aga gag aac cct cgc gcg tac ttc ttt gac ctc tgg 384

Tyr Tyr Cys Thr Arg Glu Asn Pro Arg Ala Tyr Phe Phe Asp Leu Trp

115 120 125

ggc cag gga acc ctg gtc acc gtc tcc tca 414

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

130 135

<210> 26

<211> 138

<212> PRT

<213> Homo sapiens

<400> 26

Met Asn Leu Gly Leu Ser Leu Ile Phe Leu Ala Leu Ile Leu Lys Gly

1 5 10 15

Val Gln Cys Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys

20 25 30

Pro Gly Ala Ser Val Lys Val Ser Cys Gln Ala Ser Gly Tyr Thr Phe

35 40 45

Thr Gly Asp Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu

50 55 60

Glu Tyr Leu Gly Trp Ile Asn Pro Asp Arg Gly Phe Thr Tyr Tyr Thr

65 70 75 80

Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Asn

85 90 95

Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Met

100 105 110

Tyr Tyr Cys Thr Arg Glu Asn Pro Arg Ala Tyr Phe Phe Asp Leu Trp

115 120 125

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

130 135

<210> 27

<211> 119

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VH

of MOG14 excluding signal sequence

<400> 27

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Gln Ala Ser Gly Tyr Thr Phe Thr Gly Asp

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Tyr Leu

35 40 45

Gly Trp Ile Asn Pro Asp Arg Gly Phe Thr Tyr Tyr Tyr Thr Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ser Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Glu Asn Pro Arg Ala Tyr Phe Phe Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 28

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR1 of MOG14

<400> 28

Gly Asp Tyr Ile His

1 5

<210> 29

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR2 of MOG14

<400> 29

Trp Ile Asn Pro Asp Arg Gly Phe Thr Tyr Tyr Tyr Thr Gln Lys Phe Gln

1 5 10 15

Gly

<210> 30

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

HCDR3 of MOG14

<400> 30

Glu Asn Pro Arg Ala Tyr Phe Phe Asp Leu

1 5 10

<210> 31

<211> 381

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(381)

<400> 31

atg aag ttg cct gtt agg ctg ttg gtg ctg atg ttc tgg att cct gct 48

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala

1 5 10 15

tcc agc agt gaa ata gtg ttg acg cag tct cca ggc acc ctg tct ttg 96

Ser Ser Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu

20 25 30

tct cca ggg gaa aga gcc act ctc tcc tgc agg gcc agt cag agt att 144

Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile

35 40 45

agc ggc agc tac gtg acc tgg tac cag cag aag cct ggc cag gct ccc 192

Ser Gly Ser Tyr Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

50 55 60

agg ctc ctc atc tat gct aca tcc aat agg gcc att ggc atc cca gac 240

Arg Leu Leu Ile Tyr Ala Thr Ser Asn Arg Ala Ile Gly Ile Pro Asp

65 70 75 80

aag ttc agt ggc ggt ggg tct ggg aga gac ttc act ctc acc atc aac 288

Lys Phe Ser Gly Gly Gly Ser Gly Arg Asp Phe Thr Leu Thr Ile Asn

85 90 95

aga ctg gag cct gaa gat ttt gca gtg tat tac tgt cag cag agt gtt 336

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Val

100 105 110

agt tct ccg tac act ttt ggc cag ggg acc aag gtg gaa atc aaa 381

Ser Ser Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

115 120 125

<210> 32

<211> 127

<212> PRT

<213> Homo sapiens

<400> 32

Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala

1 5 10 15

Ser Ser Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu

20 25 30

Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile

35 40 45

Ser Gly Ser Tyr Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

50 55 60

Arg Leu Leu Ile Tyr Ala Thr Ser Asn Arg Ala Ile Gly Ile Pro Asp

65 70 75 80

Lys Phe Ser Gly Gly Gly Ser Gly Arg Asp Phe Thr Leu Thr Ile Asn

85 90 95

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Val

100 105 110

Ser Ser Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

115 120 125

<210> 33

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VL

of MOG14 excluding signal sequence

<400> 33

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Gly Ser

20 25 30

Tyr Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Ala Thr Ser Asn Arg Ala Ile Gly Ile Pro Asp Lys Phe Ser

50 55 60

Gly Gly Gly Ser Gly Arg Asp Phe Thr Leu Thr Ile Asn Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Val Ser Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 34

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR1 of MOG14

<400> 34

Arg Ala Ser Gln Ser Ile Ser Gly Ser Tyr Val Thr

1 5 10

<210> 35

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR2 of MOG14

<400> 35

Ala Thr Ser Asn Arg Ala Ile

1 5

<210> 36

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

LCDR3 of MOG14

<400> 36

Gln Gln Ser Val Ser Ser Pro Tyr Thr

1 5

<210> 37

<211> 411

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: sequence of VHH of

iMOG_3Rim1_S32 including signal sequence

<220>

<221> CDS

<222> (1)..(411)

<400> 37

atg tac agg atg caa ctc ctg tct tgc att gca cta agt ctt gca ctt 48

Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu

1 5 10 15

gtc acg aat tcg cag gtg cag ctc gtg gag tct ggg gga ggc ttg gtg 96

Val Thr Asn Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

cag act ggg ggg tct ctg aga ctc tcc tgt gca gcc tct gga agc atg 144

Gln Thr Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Met

35 40 45

ttc agt acc atg ggc tgg ttc cgc cag gct cca ggg aac cag cgc gag 192

Phe Ser Thr Met Gly Trp Phe Arg Gln Ala Pro Gly Asn Gln Arg Glu

50 55 60

ttg gtc gcc att atg tca tcc ggt ggt acc gca aac tat gca gac tct 240

Leu Val Ala Ile Met Ser Ser Gly Gly Thr Ala Asn Tyr Ala Asp Ser

65 70 75 80

gtg aag ggc cga ttc acc atc tcc gga gac aac gtc aag aac acg gtg 288

Val Lys Gly Arg Phe Thr Ile Ser Gly Asp Asn Val Lys Asn Thr Val

85 90 95

act ctc caa atg aac agc ctg aat cca gag gac aca gcc gtc tat tat 336

Thr Leu Gln Met Asn Ser Leu Asn Pro Glu Asp Thr Ala Val Tyr Tyr

100 105 110

tgt agg ttt acc ggt tgg gtc aag agt tcg ttc tct acg tac tgg ggc 384

Cys Arg Phe Thr Gly Trp Val Lys Ser Ser Phe Ser Thr Tyr Tyr Trp Gly

115 120 125

cag ggg acc cag gtc acc gtc tcc tca 411

Gln Gly Thr Gln Val Thr Val Ser Ser

130 135

<210> 38

<211> 137

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic Construct

<400> 38

Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu

1 5 10 15

Val Thr Asn Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Thr Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Met

35 40 45

Phe Ser Thr Met Gly Trp Phe Arg Gln Ala Pro Gly Asn Gln Arg Glu

50 55 60

Leu Val Ala Ile Met Ser Ser Gly Gly Thr Ala Asn Tyr Ala Asp Ser

65 70 75 80

Val Lys Gly Arg Phe Thr Ile Ser Gly Asp Asn Val Lys Asn Thr Val

85 90 95

Thr Leu Gln Met Asn Ser Leu Asn Pro Glu Asp Thr Ala Val Tyr Tyr

100 105 110

Cys Arg Phe Thr Gly Trp Val Lys Ser Ser Phe Ser Thr Tyr Tyr Trp Gly

115 120 125

Gln Gly Thr Gln Val Thr Val Ser Ser

130 135

<210> 39

<211> 117

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

VHH of iMOG_3Rim1_S32 excluding signal sequence

<400> 39

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Met Phe Ser Thr Met

20 25 30

Gly Trp Phe Arg Gln Ala Pro Gly Asn Gln Arg Glu Leu Val Ala Ile

35 40 45

Met Ser Ser Gly Gly Thr Ala Asn Tyr Ala Asp Ser Val Lys Gly Arg

50 55 60

Phe Thr Ile Ser Gly Asp Asn Val Lys Asn Thr Val Thr Leu Gln Met

65 70 75 80

Asn Ser Leu Asn Pro Glu Asp Thr Ala Val Tyr Tyr Cys Arg Phe Thr

85 90 95

Gly Trp Val Lys Ser Ser Ser Phe Ser Thr Tyr Tyr Trp Gly Gln Gly Thr Gln

100 105 110

Val Thr Val Ser Ser

115

<210> 40

<211> 3

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

CDR1 of iMOG_3Rim1_S32

<400> 40

Thr Met Gly

1

<210> 41

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

CDR2 of iMOG_3Rim1_S32

<400> 41

Ile Met Ser Ser Gly Gly Thr Ala Asn Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

<210> 42

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

CDR3 of iMOG_3Rim1_S32

<400> 42

Thr Gly Trp Val Lys Ser Ser Phe Ser Thr Tyr

1 5 10

<210> 43

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer1

<400> 43

ggattcctgc ttccagcagt cagtctgccc tgactc 36

<210> 44

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer2

<400> 44

ggcggccttg ggctgaccta ggacggtcag cttggt 36

<210> 45

<211> 80

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer3

<400> 45

acgccatcac agatctgcct cttcaaaatg aagttgcctg ttaggctgtt ggtgctgatg 60

ttctggattc ctgcttccag 80

<210> 46

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer4

<400> 46

aaaaggtgtc cagtgtcagg tacagctgca gcagtc 36

<210> 47

<211> 32

<212> DNA

<213> Artificial Sequence

<220>

<223> primer5

<400> 47

gccccttggt gctagctgag gagacggtga cc 32

<210> 48

<211> 80

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer6

<400> 48

acacagaccc gtcgacccct caccatgaac ctcgggctca gtttgatttt ccttgccctc 60

attttaaaag gtgtccagtg 80

<210> 49

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer7

<400> 49

ggattcctgc ttccagcagt tcctatgtgc tgactc 36

<210> 50

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer8

<400> 50

ggcggccttg ggctgaccta ggacggtcag cttggt 36

<210> 51

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer9

<400> 51

aaaaggtgtc cagtgtcagg tgcagctgca ggagtc 36

<210> 52

<211> 32

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer10

<400> 52

gccccttggt gctagctgag gagacggtga cc 32

<210> 53

<211> 40

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer11

<400> 53

ggattcctgc ttccagcagt gaaatagtgt tgacgcagtc 40

<210> 54

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer12

<400> 54

gtgcagccac cgtacgtttg atttccacct tggtcc 36

<210> 55

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer13

<400> 55

aaaaggtgtc cagtgtcagg tgcagctggt gcaatc 36

<210> 56

<211> 32

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer14

<400> 56

gccccttggt gctagctgag gagacggtga cc 32

<210> 57

<211> 40

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer15

<400> 57

tgcacttgtc acgaattcgc aggtgcagct cgtggagtct 40

<210> 58

<211> 45

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer16

<400> 58

accatatttg gactcagatc tggccgctga ggagacggtg acctg 45

<210> 59

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer17

<400> 59

acagtctcct cagctagcac caaggggcca 30

<210> 60

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer18

<400> 60

ctgctgcagc tgtacctggg accctcctcc tccgga 36

<210> 61

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer19

<400> 61

ggaggaggag ggtcccaggt acagctgcag cag 33

<210> 62

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer20

<400> 62

aagcggccgc ctggatcctc ataggacggt cag 33

<210> 63

<211> 39

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer21

<400> 63

tcagtcataa tgtctagagg agacatccag atgacccag 39

<210> 64

<211> 37

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer22

<400> 64

gggcggcctt gggctgacct ttgatctcca ccttggt 37

<210> 65

<211> 41

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer23

<400> 65

cagcctttcc tggtatactt agtgaggtgc agttggtgga g 41

<210> 66

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer24

<400> 66

tggccccttg gtgctagccg aggagacggt gaccag 36

<210> 67

<211> 738

<212> DNA

<213> Rattus norvegicus

<220>

<221> CDS

<222> (1)..(738)

<400> 67

atg gcc ggt gtg tgg agc ctt tct ctg ccc agc tgc ctc ctg tcc ctg 48

Met Ala Gly Val Trp Ser Leu Ser Leu Pro Ser Cys Leu Leu Ser Leu

1 5 10 15

ctc ctc ctc ctc cag ttg tca cgc agc tac gca gga cag ttc aga gtg 96

Leu Leu Leu Leu Gln Leu Ser Arg Ser Tyr Ala Gly Gln Phe Arg Val

20 25 30

ata ggg cca ggg cat ccc atc cgg gct tta gtt ggg gat gaa gca gaa 144

Ile Gly Pro Gly His Pro Ile Arg Ala Leu Val Gly Asp Glu Ala Glu

35 40 45

ctg ccg tgc cgt ata tct cct ggg aag aat gcc acg ggc atg gag gtg 192

Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met Glu Val

50 55 60

ggg tgg tac cgt tct ccc ttt tca aga gtg gtt cat ctg tac cga aat 240

Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr Arg Asn

65 70 75 80

ggc aag gac caa gac gca gag caa gcg cct gaa tac cgg gga cgc aca 288

Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly Arg Thr

85 90 95

gag ctt ctg aaa gag tct atc ggc gag gga aag gtt gcc ctc agg atc 336

Glu Leu Leu Lys Glu Ser Ile Gly Glu Gly Lys Val Ala Leu Arg Ile

100 105 110

cag aac gtg agg ttc tcg gat gaa gga ggc tac aca tgc ttc ttc aga 384

Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe Phe Arg

115 120 125

gac cac tcc tac caa gaa gaa gcc gcc gtg gag ttg aaa gta gaa gat 432

Asp His Ser Tyr Gln Glu Glu Ala Ala Val Glu Leu Lys Val Glu Asp

130 135 140

ccc ttc tac tgg atc aac cct ggc gtg ctg gct ctc att gcc ctt gtg 480

Pro Phe Tyr Trp Ile Asn Pro Gly Val Leu Ala Leu Ile Ala Leu Val

145 150 155 160

cct atg ctg ctc ctg cag gtc tct gta ggc ctt gta ttc ctc ttc ctg 528

Pro Met Leu Leu Leu Gln Val Ser Val Gly Leu Val Phe Leu Phe Leu

165 170 175

cag cac aga ctg aga gga aaa ctc cgt gca gaa gtc gag aat ctc cat 576

Gln His Arg Leu Arg Gly Lys Leu Arg Ala Glu Val Glu Asn Leu His

180 185 190

cgg act ttt gat cct cac ttc ctg aga gtg ccc tgc tgg aag ata aca 624

Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys Ile Thr

195 200 205

ctg ttt gtt att gtc cct gtt ctt gga ccc ctg gtt gct ttg atc atc 672

Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu Ile Ile

210 215 220

tgc tac aac tgg ctg cac cga aga ctg gca gga cag ttt ctt gaa gag 720

Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu Glu Glu

225 230 235 240

cta aga aac ccc ttt tga 738

Leu Arg Asn Pro Phe

245

<210> 68

<211> 245

<212> PRT

<213> Rattus norvegicus

<400> 68

Met Ala Gly Val Trp Ser Leu Ser Leu Pro Ser Cys Leu Leu Ser Leu

1 5 10 15

Leu Leu Leu Leu Gln Leu Ser Arg Ser Tyr Ala Gly Gln Phe Arg Val

20 25 30

Ile Gly Pro Gly His Pro Ile Arg Ala Leu Val Gly Asp Glu Ala Glu

35 40 45

Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met Glu Val

50 55 60

Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr Arg Asn

65 70 75 80

Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly Arg Thr

85 90 95

Glu Leu Leu Lys Glu Ser Ile Gly Glu Gly Lys Val Ala Leu Arg Ile

100 105 110

Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe Phe Arg

115 120 125

Asp His Ser Tyr Gln Glu Glu Ala Ala Val Glu Leu Lys Val Glu Asp

130 135 140

Pro Phe Tyr Trp Ile Asn Pro Gly Val Leu Ala Leu Ile Ala Leu Val

145 150 155 160

Pro Met Leu Leu Leu Gln Val Ser Val Gly Leu Val Phe Leu Phe Leu

165 170 175

Gln His Arg Leu Arg Gly Lys Leu Arg Ala Glu Val Glu Asn Leu His

180 185 190

Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys Ile Thr

195 200 205

Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu Ile Ile

210 215 220

Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu Glu Glu

225 230 235 240

Leu Arg Asn Pro Phe

245

<210> 69

<211> 1167

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: sequence of rMOG-FLAG-Fc

<220>

<221> CDS

<222> (1)..(1167)

<400> 69

atg gcc ggt gtg tgg agc ctt tct ctg ccc agc tgc ctc ctg tcc ctg 48

Met Ala Gly Val Trp Ser Leu Ser Leu Pro Ser Cys Leu Leu Ser Leu

1 5 10 15

ctc ctc ctc ctc cag ttg tca cgc agc tac gca gga cag ttc aga gtg 96

Leu Leu Leu Leu Gln Leu Ser Arg Ser Tyr Ala Gly Gln Phe Arg Val

20 25 30

ata ggg cca ggg cat ccc atc cgg gct tta gtt ggg gat gaa gca gaa 144

Ile Gly Pro Gly His Pro Ile Arg Ala Leu Val Gly Asp Glu Ala Glu

35 40 45

ctg ccg tgc cgt ata tct cct ggg aag aat gcc acg ggc atg gag gtg 192

Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met Glu Val

50 55 60

ggg tgg tac cgt tct ccc ttt tca aga gtg gtt cat ctg tac cga aat 240

Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr Arg Asn

65 70 75 80

ggc aag gac caa gac gca gag caa gcg cct gaa tac cgg gga cgc aca 288

Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly Arg Thr

85 90 95

gag ctt ctg aaa gag tct atc ggc gag gga aag gtt gcc ctc agg atc 336

Glu Leu Leu Lys Glu Ser Ile Gly Glu Gly Lys Val Ala Leu Arg Ile

100 105 110

cag aac gtg agg ttc tcg gat gaa gga ggc tac aca tgc ttc ttc aga 384

Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe Phe Arg

115 120 125

gac cac tcc tac caa gaa gaa gcc gcc gtg gag ttg aaa gta gaa gat 432

Asp His Ser Tyr Gln Glu Glu Ala Ala Val Glu Leu Lys Val Glu Asp

130 135 140

ccc ttc tac tgg tct aga gca gac tac aag gac gac gat gac aag act 480

Pro Phe Tyr Trp Ser Arg Ala Asp Tyr Lys Asp Asp Asp Asp Lys Thr

145 150 155 160

agt gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc ctg 528

Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

165 170 175

ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc ctc 576

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

180 185 190

atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg agc 624

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

195 200 205

cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag 672

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

210 215 220

gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc acg 720

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

225 230 235 240

tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aat 768

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

245 250 255

ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc ccc 816

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

260 265 270

atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca cag 864

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

275 280 285

gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag gtc 912

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

290 295 300

agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc gtg 960

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

305 310 315 320

gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct 1008

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

325 330 335

ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc acc 1056

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

340 345 350

gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc gtg 1104

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

355 360 365

atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc ctg 1152

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

370 375 380

tct ccg ggt aaa tga 1167

Ser Pro Gly Lys

385

<210> 70

<211> 388

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic Construct

<400> 70

Met Ala Gly Val Trp Ser Leu Ser Leu Pro Ser Cys Leu Leu Ser Leu

1 5 10 15

Leu Leu Leu Leu Gln Leu Ser Arg Ser Tyr Ala Gly Gln Phe Arg Val

20 25 30

Ile Gly Pro Gly His Pro Ile Arg Ala Leu Val Gly Asp Glu Ala Glu

35 40 45

Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met Glu Val

50 55 60

Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr Arg Asn

65 70 75 80

Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly Arg Thr

85 90 95

Glu Leu Leu Lys Glu Ser Ile Gly Glu Gly Lys Val Ala Leu Arg Ile

100 105 110

Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe Phe Arg

115 120 125

Asp His Ser Tyr Gln Glu Glu Ala Ala Val Glu Leu Lys Val Glu Asp

130 135 140

Pro Phe Tyr Trp Ser Arg Ala Asp Tyr Lys Asp Asp Asp Asp Lys Thr

145 150 155 160

Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

165 170 175

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

180 185 190

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

195 200 205

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

210 215 220

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

225 230 235 240

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

245 250 255

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

260 265 270

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

275 280 285

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

290 295 300

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

305 310 315 320

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

325 330 335

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

340 345 350

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

355 360 365

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

370 375 380

Ser Pro Gly Lys

385

<210> 71

<211> 1137

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: sequence of rMOG-GST

<220>

<221> CDS

<222> (1)..(1137)

<400> 71

atg gcc ggt gtg tgg agc ctt tct ctg ccc agc tgc ctc ctg tcc ctg 48

Met Ala Gly Val Trp Ser Leu Ser Leu Pro Ser Cys Leu Leu Ser Leu

1 5 10 15

ctc ctc ctc ctc cag ttg tca cgc agc tac gca gga cag ttc aga gtg 96

Leu Leu Leu Leu Gln Leu Ser Arg Ser Tyr Ala Gly Gln Phe Arg Val

20 25 30

ata ggg cca ggg cat ccc atc cgg gct tta gtt ggg gat gaa gca gaa 144

Ile Gly Pro Gly His Pro Ile Arg Ala Leu Val Gly Asp Glu Ala Glu

35 40 45

ctg ccg tgc cgt ata tct cct ggg aag aat gcc acg ggc atg gag gtg 192

Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met Glu Val

50 55 60

ggg tgg tac cgt tct ccc ttt tca aga gtg gtt cat ctg tac cga aat 240

Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr Arg Asn

65 70 75 80

ggc aag gac caa gac gca gag caa gcg cct gaa tac cgg gga cgc aca 288

Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly Arg Thr

85 90 95

gag ctt ctg aaa gag tct atc ggc gag gga aag gtt gcc ctc agg atc 336

Glu Leu Leu Lys Glu Ser Ile Gly Glu Gly Lys Val Ala Leu Arg Ile

100 105 110

cag aac gtg agg ttc tcg gat gaa gga ggc tac aca tgc ttc ttc aga 384

Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe Phe Arg

115 120 125

gac cac tcc tac caa gaa gaa gcc gcc gtg gag ttg aaa gta gaa gat 432

Asp His Ser Tyr Gln Glu Glu Ala Ala Val Glu Leu Lys Val Glu Asp

130 135 140

ccc ttc tac tgg ggt acc ctg gaa gtt ctg ttc cag ggg ccc atg tcc 480

Pro Phe Tyr Trp Gly Thr Leu Glu Val Leu Phe Gln Gly Pro Met Ser

145 150 155 160

cct ata cta ggt tat tgg aaa att aag ggc ctt gtg caa ccc act cga 528

Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu Val Gln Pro Thr Arg

165 170 175

ctt ctt ttg gaa tat ctt gaa gaa aaa tat gaa gag cat ttg tat gag 576

Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu

180 185 190

cgc gat gaa ggt gat aaa tgg cga aac aaa aag ttt gaa ttg ggt ttg 624

Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu Gly Leu

195 200 205

gag ttt ccc aat ctt cct tat tat att gat ggt gat gtt aaa tta aca 672

Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly Asp Val Lys Leu Thr

210 215 220

cag tct atg gcc atc ata cgt tat ata gct gac aag cac aac atg ttg 720

Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp Lys His Asn Met Leu

225 230 235 240

ggt ggt tgt cca aaa gag cgt gca gag att tca atg ctt gaa gga gcg 768

Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser Met Leu Glu Gly Ala

245 250 255

gtt ttg gat att aga tac ggt gtt tcg aga att gca tat agt aaa gac 816

Val Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr Ser Lys Asp

260 265 270

ttt gaa act ctc aaa gtt gat ttt ctt agc aag cta cct gaa atg ctg 864

Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu Met Leu

275 280 285

aaa atg ttc gaa gat cgt tta tgt cat aaa aca tat tta aat ggt gat 912

Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn Gly Asp

290 295 300

cat gta acc cat cct gac ttc atg ttg tat gac gct ctt gat gtt gtt 960

His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp Val Val

305 310 315 320

tta tac atg gac cca atg tgc ctg gat gcg ttc cca aaa tta gtt tgt 1008

Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu Val Cys

325 330 335

ttt aaa aaa cgt att gaa gct atc cca caa att gat aag tac ttg aaa 1056

Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile Asp Lys Tyr Leu Lys

340 345 350

tcc agc aag tat ata gca tgg cct ttg cag ggc tgg caa gcc acg ttt 1104

Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly Trp Gln Ala Thr Phe

355 360 365

ggt ggt ggc gac cat cct cca aaa tcg gat tga 1137

Gly Gly Gly Asp His Pro Pro Lys Ser Asp

370 375

<210> 72

<211> 378

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic Construct

<400> 72

Met Ala Gly Val Trp Ser Leu Ser Leu Pro Ser Cys Leu Leu Ser Leu

1 5 10 15

Leu Leu Leu Leu Gln Leu Ser Arg Ser Tyr Ala Gly Gln Phe Arg Val

20 25 30

Ile Gly Pro Gly His Pro Ile Arg Ala Leu Val Gly Asp Glu Ala Glu

35 40 45

Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met Glu Val

50 55 60

Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr Arg Asn

65 70 75 80

Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly Arg Thr

85 90 95

Glu Leu Leu Lys Glu Ser Ile Gly Glu Gly Lys Val Ala Leu Arg Ile

100 105 110

Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe Phe Arg

115 120 125

Asp His Ser Tyr Gln Glu Glu Ala Ala Val Glu Leu Lys Val Glu Asp

130 135 140

Pro Phe Tyr Trp Gly Thr Leu Glu Val Leu Phe Gln Gly Pro Met Ser

145 150 155 160

Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu Val Gln Pro Thr Arg

165 170 175

Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu

180 185 190

Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu Gly Leu

195 200 205

Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly Asp Val Lys Leu Thr

210 215 220

Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp Lys His Asn Met Leu

225 230 235 240

Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser Met Leu Glu Gly Ala

245 250 255

Val Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr Ser Lys Asp

260 265 270

Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu Met Leu

275 280 285

Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn Gly Asp

290 295 300

His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp Val Val

305 310 315 320

Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu Val Cys

325 330 335

Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile Asp Lys Tyr Leu Lys

340 345 350

Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly Trp Gln Ala Thr Phe

355 360 365

Gly Gly Gly Asp His Pro Pro Lys Ser Asp

370 375

<210> 73

<211> 744

<212> DNA

<213> Mus musculus

<220>

<221> CDS

<222> (1)..(744)

<400> 73

atg gcc tgt ttg tgg agc ttc tct tgg ccc agc tgc ttc ctc tcc ctt 48

Met Ala Cys Leu Trp Ser Phe Ser Trp Pro Ser Cys Phe Leu Ser Leu

1 5 10 15

ctc ctc ctc ctt ctc ctc cag ttg tca tgc agc tat gca gga caa ttc 96

Leu Leu Leu Leu Leu Leu Gln Leu Ser Cys Ser Tyr Ala Gly Gln Phe

20 25 30

aga gtg ata gga cca ggg tat ccc atc cgg gct tta gtt ggg gat gaa 144

Arg Val Ile Gly Pro Gly Tyr Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

gca gag ctg ccg tgc cgc atc tct cct ggg aaa aat gcc acg ggc atg 192

Ala Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

gag gtg ggt tgg tac cgt tct ccc ttc tca aga gtg gtt cac ctc tac 240

Glu Val Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

cga aat ggc aag gac caa gat gca gag caa gca cct gaa tac cgg gga 288

Arg Asn Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly

85 90 95

cgc aca gag ctt ctg aaa gag act atc agt gag gga aag gtt acc ctt 336

Arg Thr Glu Leu Leu Lys Glu Thr Ile Ser Glu Gly Lys Val Thr Leu

100 105 110

agg att cag aac gtg aga ttc tca gat gaa gga ggc tac acc tgc ttc 384

Arg Ile Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe

115 120 125

ttc aga gac cac tct tac caa gaa gag gca gca atg gag ttg aaa gtg 432

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

gaa gat ccc ttc tat tgg gtc aac ccc ggt gtg ctg act ctc atc gca 480

Glu Asp Pro Phe Tyr Trp Val Asn Pro Gly Val Leu Thr Leu Ile Ala

145 150 155 160

ctt gtg cct acg atc ctc ctg cag gtc tct gta ggc ctt gta ttc ctc 528

Leu Val Pro Thr Ile Leu Leu Gln Val Ser Val Gly Leu Val Phe Leu

165 170 175

ttc ctg cag cac aga ctg aga gga aaa ctt cgt gca gaa gta gag aat 576

Phe Leu Gln His Arg Leu Arg Gly Lys Leu Arg Ala Glu Val Glu Asn

180 185 190

ctc cat cgg act ttt gat cct cac ttc ctg agg gtg ccc tgc tgg aag 624

Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys

195 200 205

ata aca ctg ttt gtt att gtg cct gtt ctt gga ccc ctg gtt gcc ttg 672

Ile Thr Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu

210 215 220

atc atc tgc tac aac tgg ctg cac cga aga ctg gca gga cag ttt ctt 720

Ile Ile Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu

225 230 235 240

gaa gag cta aga aac ccc ttt tga 744

Glu Glu Leu Arg Asn Pro Phe

245

<210> 74

<211> 247

<212> PRT

<213> Mus musculus

<400> 74

Met Ala Cys Leu Trp Ser Phe Ser Trp Pro Ser Cys Phe Leu Ser Leu

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Leu Ser Cys Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Gly Tyr Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Ala Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Glu Thr Ile Ser Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Asn Pro Gly Val Leu Thr Leu Ile Ala

145 150 155 160

Leu Val Pro Thr Ile Leu Leu Gln Val Ser Val Gly Leu Val Phe Leu

165 170 175

Phe Leu Gln His Arg Leu Arg Gly Lys Leu Arg Ala Glu Val Glu Asn

180 185 190

Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys

195 200 205

Ile Thr Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu

210 215 220

Ile Ile Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu

225 230 235 240

Glu Glu Leu Arg Asn Pro Phe

245

<210> 75

<211> 744

<212> DNA

<213> Macaca fascicularis

<220>

<221> CDS

<222> (1)..(744)

<400> 75

atg gca agc tta tca aga ccc tct ctg ccc agc tgc ctc tgc tcc ttc 48

Met Ala Ser Leu Ser Arg Pro Ser Leu Pro Ser Cys Leu Cys Ser Phe

1 5 10 15

ctc ctc ctc ctg ctc ctc caa gtg tct tcc agc tac gca gga cag ttc 96

Leu Leu Leu Leu Leu Leu Gln Val Ser Ser Ser Tyr Ala Gly Gln Phe

20 25 30

aga gtg ata gga cca aga caa cct atc cgg gct ctg gtc ggt gat gaa 144

Arg Val Ile Gly Pro Arg Gln Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

gtg gaa ttg cca tgt cgc ata tct cct ggg aag aac gct aca ggc atg 192

Val Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

gaa gtg gga tgg tac cgg ccc ccc ttc tct agg gtg gtt cat ctc tac 240

Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

aga aat ggc agg gac caa gat gga gag caa gca cct gaa tat cgg ggc 288

Arg Asn Gly Arg Asp Gln Asp Gly Glu Gln Ala Pro Glu Tyr Arg Gly

85 90 95

cgg aca gag ctg ctg aaa gac gct att ggt gag gga aag gtg act ctc 336

Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys Val Thr Leu

100 105 110

agg atc cgg aat gta agg ttc tca gat gaa gga ggt ttc acc tgc ttc 384

Arg Ile Arg Asn Val Arg Phe Ser Asp Glu Gly Gly Phe Thr Cys Phe

115 120 125

ttc cga gat cat tct tac caa gag gag gca gca ata gaa ttg aaa gtg 432

Phe Arg Asp His Ser Tyr Gln Glu Glu Ala Ala Ile Glu Leu Lys Val

130 135 140

gaa gat ccc ttc tac tgg gtc agc cct gca gtg ctg gtt ctc ctc gcg 480

Glu Asp Pro Phe Tyr Trp Val Ser Pro Ala Val Leu Val Leu Leu Ala

145 150 155 160

gtg ctg cct gtg ctc ctt ctg cag atc act gtc ggc ctc gtc ttc ctc 528

Val Leu Pro Val Leu Leu Leu Gln Ile Thr Val Gly Leu Val Phe Leu

165 170 175

tgc ctg cag tat aga ctg aga gga aaa ctt cga gca gag ata gag aat 576

Cys Leu Gln Tyr Arg Leu Arg Gly Lys Leu Arg Ala Glu Ile Glu Asn

180 185 190

ctc cac cgg act ttt gat ccc cac ttt ctg agg gtg ccc tgc tgg aag 624

Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys

195 200 205

ata acc ctg ttt gta att gtg ccg gtt ctt gga ccc ctg gtt gcc ttg 672

Ile Thr Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu

210 215 220

atc atc tgc tac aac tgg cta cat cga aga cta gca ggg caa ttc ctt 720

Ile Ile Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu

225 230 235 240

gaa gag cta aga aac cct ttc tga 744

Glu Glu Leu Arg Asn Pro Phe

245

<210> 76

<211> 247

<212> PRT

<213> Macaca fascicularis

<400> 76

Met Ala Ser Leu Ser Arg Pro Ser Leu Pro Ser Cys Leu Cys Ser Phe

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Val Ser Ser Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Arg Gln Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Val Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Arg Asp Gln Asp Gly Glu Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Arg Asn Val Arg Phe Ser Asp Glu Gly Gly Phe Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Glu Ala Ala Ile Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Ser Pro Ala Val Leu Val Leu Leu Ala

145 150 155 160

Val Leu Pro Val Leu Leu Leu Gln Ile Thr Val Gly Leu Val Phe Leu

165 170 175

Cys Leu Gln Tyr Arg Leu Arg Gly Lys Leu Arg Ala Glu Ile Glu Asn

180 185 190

Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys

195 200 205

Ile Thr Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu

210 215 220

Ile Ile Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu

225 230 235 240

Glu Glu Leu Arg Asn Pro Phe

245

<210> 77

<211> 744

<212> DNA

<213> Homo sapiens

<220>

<221> CDS

<222> (1)..(744)

<400> 77

atg gca agc tta tca aga ccc tct ctg ccc agc tgc ctc tgc tcc ttc 48

Met Ala Ser Leu Ser Arg Pro Ser Leu Pro Ser Cys Leu Cys Ser Phe

1 5 10 15

ctc ctc ctc ctc ctc ctc caa gtg tct tcc agc tat gca ggg cag ttc 96

Leu Leu Leu Leu Leu Leu Gln Val Ser Ser Ser Tyr Ala Gly Gln Phe

20 25 30

aga gtg ata gga cca aga cac cct atc cgg gct ctg gtc ggg gat gaa 144

Arg Val Ile Gly Pro Arg His Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

gtg gaa ttg cca tgt cgc ata tct cct ggg aag aac gct aca ggc atg 192

Val Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

gag gtg ggg tgg tac cgc ccc ccc ttc tct agg gtg gtt cat ctc tac 240

Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

aga aat ggc aag gac caa gat gga gac cag gca cct gaa tat cgg ggc 288

Arg Asn Gly Lys Asp Gln Asp Gly Asp Gln Ala Pro Glu Tyr Arg Gly

85 90 95

cgg aca gag ctg ctg aaa gat gct att ggt gag gga aag gtg act ctc 336

Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys Val Thr Leu

100 105 110

agg atc cgg aat gta agg ttc tca gat gaa gga ggt ttc acc tgc ttc 384

Arg Ile Arg Asn Val Arg Phe Ser Asp Glu Gly Gly Phe Thr Cys Phe

115 120 125

ttc cga gat cat tct tac caa gag gag gca gca atg gaa ttg aaa gta 432

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

gaa gat cct ttc tac tgg gtg agc cct gga gtg ctg gtt ctc ctc gcg 480

Glu Asp Pro Phe Tyr Trp Val Ser Pro Gly Val Leu Val Leu Leu Ala

145 150 155 160

gtg ctg cct gtg ctc ctc ctg cag atc act gtt ggc ctc gtc ttc ctc 528

Val Leu Pro Val Leu Leu Leu Gln Ile Thr Val Gly Leu Val Phe Leu

165 170 175

tgc ctg cag tac aga ctg aga gga aaa ctt cga gca gag ata gag aat 576

Cys Leu Gln Tyr Arg Leu Arg Gly Lys Leu Arg Ala Glu Ile Glu Asn

180 185 190

ctc cac cgg act ttt gat ccc cac ttt ctg agg gtg ccc tgc tgg aag 624

Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys

195 200 205

ata acc ctg ttt gta att gtg ccg gtt ctt gga ccc ttg gtt gcc ttg 672

Ile Thr Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu

210 215 220

atc atc tgc tac aac tgg cta cat cga aga cta gca ggg caa ttc ctt 720

Ile Ile Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu

225 230 235 240

gaa gag cta cga aat ccc ttc tga 744

Glu Glu Leu Arg Asn Pro Phe

245

<210> 78

<211> 247

<212> PRT

<213> Homo sapiens

<400> 78

Met Ala Ser Leu Ser Arg Pro Ser Leu Pro Ser Cys Leu Cys Ser Phe

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Val Ser Ser Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Arg His Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Val Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Lys Asp Gln Asp Gly Asp Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Arg Asn Val Arg Phe Ser Asp Glu Gly Gly Phe Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Ser Pro Gly Val Leu Val Leu Leu Ala

145 150 155 160

Val Leu Pro Val Leu Leu Leu Gln Ile Thr Val Gly Leu Val Phe Leu

165 170 175

Cys Leu Gln Tyr Arg Leu Arg Gly Lys Leu Arg Ala Glu Ile Glu Asn

180 185 190

Leu His Arg Thr Phe Asp Pro His Phe Leu Arg Val Pro Cys Trp Lys

195 200 205

Ile Thr Leu Phe Val Ile Val Pro Val Leu Gly Pro Leu Val Ala Leu

210 215 220

Ile Ile Cys Tyr Asn Trp Leu His Arg Arg Leu Ala Gly Gln Phe Leu

225 230 235 240

Glu Glu Leu Arg Asn Pro Phe

245

<210> 79

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer25

<400> 79

gagacgccat cacagatcat cccaccatgt aca 33

<210> 80

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer26

<400> 80

gtaaccgtta acggatcctc atttacccag aga 33

<210> 81

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer27

<400> 81

accaaggtgg agatcaaacg tacggtggct gcacca 36

<210> 82

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer28

<400> 82

cggccacacg ttgaattctc aacactctcc cctgtt 36

<210> 83

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer29

<400> 83

gattcctgct tccagcagtc agtttgtgct ttctca 36

<210> 84

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer30

<400> 84

ggcggccttg ggctgaccta ggacagtgag cttggt 36

<210> 85

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer31

<400> 85

ttaaaaggtg tccagtgtga ggtgcagctg gtggaa 36

<210> 86

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer32

<400> 86

tggccccttg gtgctagctg aggagactgt gaccat 36

<210> 87

<211> 38

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer33

<400> 87

tcagtcataa tgtctagagg acagtttgtg ctttctca 38

<210> 88

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer34

<400> 88

cggccacacg ttgaattctc atgaacattc tgtagg 36

<210> 89

<211> 41

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer35

<400> 89

cagcctttcc tggtatactt agtgaggtgc agctggtgga a 41

<210> 90

<211> 30

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer36

<400> 90

acagtctcct cagctagcac caaggggcca 30

<210> 91

<211> 35

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer37

<400> 91

tccaccagct gcacctcgga ccctcctcct ccgga 35

<210> 92

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer38

<400> 92

ggaggaggag ggtccgaggt gcagctggtg gaa 33

<210> 93

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer39

<400> 93

aagcggccgc ctggatcctc ataggacagt gagctt 36

<210> 94

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer40

<400> 94

agacgccatc agatctgc ctcttcaaaa tga 33

<210> 95

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer41

<400> 95

tggtgcagcc accgtacgtt tgatttccag cttggt 36

<210> 96

<211> 33

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer42

<400> 96

gaacacagac ccgtcgaccc ctcaccatga acc 33

<210> 97

<211> 36

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: primer43

<400> 97

tggccccttg gtgctagcgg aggagactgt gagagt 36

<210> 98

<211> 2640

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: sequence of hHER2-GST

<220>

<221> CDS

<222> (1)..(2640)

<400> 98

atg gag ctg gcg gcc ttg tgc cgc tgg ggg ctc ctc ctc gcc ctc ttg 48

Met Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu Ala Leu Leu

1 5 10 15

ccc ccc gga gcc gcg agc acc caa gtg tgc acc ggc aca gac atg aag 96

Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly Thr Asp Met Lys

20 25 30

ctg cgg ctc cct gcc agt ccc gag acc cac ctg gac atg ctc cgc cac 144

Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg His

35 40 45

ctc tac cag ggc tgc cag gtg gtg cag gga aac ctg gaa ctc acc tac 192

Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr

50 55 60

ctg ccc acc aat gcc agc ctg tcc ttc ctg cag gat atc cag gag gtg 240

Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val

65 70 75 80

cag ggc tac gtg ctc atc gct cac aac caa gtg agg cag gtc cca ctg 288

Gln Gly Tyr Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu

85 90 95

cag agg ctg cgg att gtg cga ggc acc cag ctc ttt gag gac aac tat 336

Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr

100 105 110

gcc ctg gcc gtg cta gac aat gga gac ccg ctg aac aat acc acc cct 384

Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro

115 120 125

gtc aca ggg gcc tcc cca gga ggc ctg cgg gag ctg cag ctt cga agc 432

Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser

130 135 140

ctc aca gag atc ttg aaa gga ggg gtc ttg atc cag cgg aac ccc cag 480

Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln

145 150 155 160

ctc tgc tac cag gac acg att ttg tgg aag gac atc ttc cac aag aac 528

Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys Asn

165 170 175

aac cag ctg gct ctc aca ctg ata gac acc aac cgc tct cgg gcc tgc 576

Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys

180 185 190

cac ccc tgt tct ccg atg tgt aag ggc tcc cgc tgc tgg gga gag agt 624

His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser

195 200 205

tct gag gat tgt cag agc ctg acg cgc act gtc tgt gcc ggt ggc tgt 672

Ser Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys

210 215 220

gcc cgc tgc aag ggg cca ctg ccc act gac tgc tgc cat gag cag tgt 720

Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln Cys

225 230 235 240

gct gcc ggc tgc acg ggc ccc aag cac tct gac tgc ctg gcc tgc ctc 768

Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu

245 250 255

cac ttc aac cac agt ggc atc tgt gag ctg cac tgc cca gcc ctg gtc 816

His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val

260 265 270

acc tac aac aca gac acg ttt gag tcc atg ccc aat ccc gag ggc cgg 864

Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg

275 280 285

tat aca ttc ggc gcc agc tgt gtg act gcc tgt ccc tac aac tac ctt 912

Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu

290 295 300

tct acg gac gtg gga tcc tgc acc ctc gtc tgc ccc ctg cac aac caa 960

Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn Gln

305 310 315 320

gag gtg aca gca gag gat gga aca cag cgg tgt gag aag tgc agc aag 1008

Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys

325 330 335

ccc tgt gcc cga gtg tgc tat ggt ctg ggc atg gag cac ttg cga gag 1056

Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu

340 345 350

gtg agg gca gtt acc agt gcc aat atc cag gag ttt gct ggc tgc aag 1104

Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys

355 360 365

aag atc ttt ggg agc ctg gca ttt ctg ccg gag agc ttt gat ggg gac 1152

Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp

370 375 380

cca gcc tcc aac act gcc ccg ctc cag cca gag cag ctc caa gtg ttt 1200

Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln Val Phe

385 390 395 400

gag act ctg gaa gag atc aca ggt tac cta tac atc tca gca tgg ccg 1248

Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro

405 410 415

gac agc ctg cct gac ctc agc gtc ttc cag aac ctg caa gta atc cgg 1296

Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln Val Ile Arg

420 425 430

gga cga att ctg cac aat ggc gcc tac tcg ctg acc ctg caa ggg ctg 1344

Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu

435 440 445

ggc atc agc tgg ctg ggg ctg cgc tca ctg agg gaa ctg ggc agt gga 1392

Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly

450 455 460

ctg gcc ctc atc cac cat aac acc cac ctc tgc ttc gtg cac acg gtg 1440

Leu Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His Thr Val

465 470 475 480

ccc tgg gac cag ctc ttt cgg aac ccg cac caa gct ctg ctc cac act 1488

Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Thr

485 490 495

gcc aac cgg cca gag gac gag tgt gtg ggc gag ggc ctg gcc tgc cac 1536

Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala Cys His

500 505 510

cag ctg tgc gcc cga ggg cac tgc tgg ggt cca ggg ccc acc cag tgt 1584

Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys

515 520 525

gtc aac tgc agc cag ttc ctt cgg ggc cag gag tgc gtg gag gaa tgc 1632

Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys

530 535 540

cga gta ctg cag ggg ctc ccc agg gag tat gtg aat gcc agg cac tgt 1680

Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg His Cys

545 550 555 560

ttg ccg tgc cac cct gag tgt cag ccc cag aat ggc tca gtg acc tgt 1728

Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys

565 570 575

ttt gga ccg gag gct gac cag tgt gtg gcc tgt gcc cac tat aag gac 1776

Phe Gly Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp

580 585 590

cct ccc ttc tgc gtg gcc cgc tgc ccc agc ggt gtg aaa cct gac ctc 1824

Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu

595 600 605

tcc tac atg ccc atc tgg aag ttt cca gat gag gag ggc gca tgc cag 1872

Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln

610 615 620

cct tgc ccc atc aac tgc acc cac tcc tgt gtg gac ctg gat gac aag 1920

Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp Lys

625 630 635 640

ggc tgc ccc gcc gag cag aga gcc agc ggt acc ctg gaa gtt ctg ttc 1968

Gly Cys Pro Ala Glu Gln Arg Ala Ser Gly Thr Leu Glu Val Leu Phe

645 650 655

cag ggg ccc atg tcc cct ata cta ggt tat tgg aaa att aag ggc ctt 2016

Gln Gly Pro Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu

660 665 670

gtg caa ccc act cga ctt ctt ttg gaa tat ctt gaa gaa aaa tat gaa 2064

Val Gln Pro Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu

675 680 685

gag cat ttg tat gag cgc gat gaa ggt gat aaa tgg cga aac aaa aag 2112

Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys

690 695 700

ttt gaa ttg ggt ttg gag ttt ccc aat ctt cct tat tat att gat ggt 2160

Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly

705 710 715 720

gat gtt aaa tta aca cag tct atg gcc atc ata cgt tat ata gct gac 2208

Asp Val Lys Leu Thr Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp

725 730 735

aag cac aac atg ttg ggt ggt tgt cca aaa gag cgt gca gag att tca 2256

Lys His Asn Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser

740 745 750

atg ctt gaa gga gcg gtt ttg gat att aga tac ggt gtt tcg aga att 2304

Met Leu Glu Gly Ala Val Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile

755 760 765

gca tat agt aaa gac ttt gaa act ctc aaa gtt gat ttt ctt agc aag 2352

Ala Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys

770 775 780

cta cct gaa atg ctg aaa atg ttc gaa gat cgt tta tgt cat aaa aca 2400

Leu Pro Glu Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr

785 790 795 800

tat tta aat ggt gat cat gta acc cat cct gac ttc atg ttg tat gac 2448

Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp

805 810 815

gct ctt gat gtt gtt tta tac atg gac cca atg tgc ctg gat gcg ttc 2496

Ala Leu Asp Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe

820 825 830

cca aaa tta gtt tgt ttt aaa aaa cgt att gaa gct atc cca caa att 2544

Pro Lys Leu Val Cys Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile

835 840 845

gat aag tac ttg aaa tcc agc aag tat ata gca tgg cct ttg cag ggc 2592

Asp Lys Tyr Leu Lys Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly

850 855 860

tgg caa gcc acg ttt ggt ggt ggc gac cat cct cca aaa tcg gat tga 2640

Trp Gln Ala Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp

865 870 875

<210> 99

<211> 879

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic Construct

<400> 99

Met Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu Ala Leu Leu

1 5 10 15

Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly Thr Asp Met Lys

20 25 30

Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg His

35 40 45

Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr

50 55 60

Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val

65 70 75 80

Gln Gly Tyr Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu

85 90 95

Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr

100 105 110

Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro

115 120 125

Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser

130 135 140

Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln

145 150 155 160

Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys Asn

165 170 175

Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys

180 185 190

His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser

195 200 205

Ser Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys

210 215 220

Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln Cys

225 230 235 240

Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu

245 250 255

His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val

260 265 270

Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg

275 280 285

Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu

290 295 300

Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn Gln

305 310 315 320

Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys

325 330 335

Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu

340 345 350

Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys

355 360 365

Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp

370 375 380

Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln Val Phe

385 390 395 400

Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro

405 410 415

Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln Val Ile Arg

420 425 430

Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu

435 440 445

Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly

450 455 460

Leu Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His Thr Val

465 470 475 480

Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Thr

485 490 495

Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala Cys His

500 505 510

Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys

515 520 525

Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys

530 535 540

Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg His Cys

545 550 555 560

Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys

565 570 575

Phe Gly Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp

580 585 590

Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu

595 600 605

Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln

610 615 620

Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp Lys

625 630 635 640

Gly Cys Pro Ala Glu Gln Arg Ala Ser Gly Thr Leu Glu Val Leu Phe

645 650 655

Gln Gly Pro Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu

660 665 670

Val Gln Pro Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu

675 680 685

Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys

690 695 700

Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly

705 710 715 720

Asp Val Lys Leu Thr Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp

725 730 735

Lys His Asn Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser

740 745 750

Met Leu Glu Gly Ala Val Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile

755 760 765

Ala Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys

770 775 780

Leu Pro Glu Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr

785 790 795 800

Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp

805 810 815

Ala Leu Asp Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe

820 825 830

Pro Lys Leu Val Cys Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile

835 840 845

Asp Lys Tyr Leu Lys Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly

850 855 860

Trp Gln Ala Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp

865 870 875

<210> 100

<211> 1182

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of

hMOG-FLAG-Fc including signal sequence

<400> 100

atggcaagct tatcaagacc ctctctgccc agctgcctct gctccttcct cctcctcctc 60

ctcctccaag tgtcttccag ctatgcaggg cagttcagag tgataggacc aagacaccct 120

atccgggctc tggtcgggga tgaagtggaa ttgccatgtc gcatatctcc tgggaagaac 180

gctacaggca tggaggtggg gtggtaccgc ccccccttct ctagggtggt tcatctctac 240

agaaatggca aggaccaaga tggagaccag gcacctgaat atcggggccg gacagagctg 300

ctgaaagatg ctattggtga gggaaaggtg actctcagga tccggaatgt aaggttctca 360

gatgaaggag gtttcacctg cttcttccga gatcattctt accaagagga ggcagcaatg 420

gaattgaaag tagaagatcc tttctactgg gtgagccctg gatctagagc agactacaag 480

gacgacgatg acaagactag tgacaaaact cacacatgcc caccgtgccc agcacctgaa 540

ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc 600

tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc 660

aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag 720

gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg 780

ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag 840

aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca 900

tcccgggatg agctgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat 960

cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc 1020

acgcctcccg tgctggactc cgacggctcc ttcttcctct acaagcaagct caccgtggac 1080

aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga ggctctgcac 1140

aaccactaca cgcagaagag cctctccctg tctccgggta aa 1182

<210> 101

<211> 394

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

hMOG-FLAG-Fc including signal sequence

<400> 101

Met Ala Ser Leu Ser Arg Pro Ser Leu Pro Ser Cys Leu Cys Ser Phe

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Val Ser Ser Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Arg His Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Val Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Lys Asp Gln Asp Gly Asp Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Arg Asn Val Arg Phe Ser Asp Glu Gly Gly Phe Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Ser Pro Gly Ser Arg Ala Asp Tyr Lys

145 150 155 160

Asp Asp Asp Asp Lys Thr Ser Asp Lys Thr His Thr Cys Pro Pro Cys

165 170 175

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

180 185 190

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

195 200 205

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

210 215 220

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

225 230 235 240

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

245 250 255

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

260 265 270

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

275 280 285

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

290 295 300

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

305 310 315 320

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

325 330 335

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

340 345 350

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

355 360 365

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

370 375 380

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

385 390

<210> 102

<211> 1182

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence of

mMOG-FLAG-Fc including signal sequence

<400> 102

atggcctgtt tgtggagctt ctttggccc agctgcttcc tctcccttct cctcctcctt 60

ctcctccagt tgtcatgcag ctatgcagga caattcagag tgataggacc agggtatccc 120

atccgggctt tagttgggga tgaagcagag ctgccgtgcc gcatctctcc tgggaaaaat 180

gccacgggca tggaggtggg ttggtaccgt tctcccttct caagagtggt tcacctctac 240

cgaaatggca aggaccaaga tgcagagcaa gcacctgaat accggggacg cacagagctt 300

ctgaaagaga ctatcagtga gggaaaggtt acccttagga ttcagaacgt gagattctca 360

gatgaaggag gctacacctg cttcttcaga gaccactctt accaagaaga ggcagcaatg 420

gagttgaaag tggaagatcc cttctattgg gtcaaccccg gttctagagc agactacaag 480

gacgacgatg acaagactag tgacaaaact cacacatgcc caccgtgccc agcacctgaa 540

ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc 600

tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc 660

aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag 720

gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg 780

ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag 840

aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca 900

tcccgggatg agctgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat 960

cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc 1020

acgcctcccg tgctggactc cgacggctcc ttcttcctct acaagcaagct caccgtggac 1080

aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga ggctctgcac 1140

aaccactaca cgcagaagag cctctccctg tctccgggta aa 1182

<210> 103

<211> 394

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

mMOG-FLAG-Fc including signal sequence

<400> 103

Met Ala Cys Leu Trp Ser Phe Ser Trp Pro Ser Cys Phe Leu Ser Leu

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Leu Ser Cys Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Gly Tyr Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Ala Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Glu Thr Ile Ser Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Asn Pro Gly Ser Arg Ala Asp Tyr Lys

145 150 155 160

Asp Asp Asp Asp Lys Thr Ser Asp Lys Thr His Thr Cys Pro Pro Cys

165 170 175

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

180 185 190

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

195 200 205

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

210 215 220

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

225 230 235 240

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

245 250 255

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

260 265 270

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

275 280 285

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

290 295 300

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

305 310 315 320

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

325 330 335

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

340 345 350

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

355 360 365

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

370 375 380

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

385 390

<210> 104

<211> 1152

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of hMOG-GST

including signal sequence

<400> 104

atggcaagct tatcaagacc ctctctgccc agctgcctct gctccttcct cctcctcctc 60

ctcctccaag tgtcttccag ctatgcaggg cagttcagag tgataggacc aagacaccct 120

atccgggctc tggtcgggga tgaagtggaa ttgccatgtc gcatatctcc tgggaagaac 180

gctacaggca tggaggtggg gtggtaccgc ccccccttct ctagggtggt tcatctctac 240

agaaatggca aggaccaaga tggagaccag gcacctgaat atcggggccg gacagagctg 300

ctgaaagatg ctattggtga gggaaaggtg actctcagga tccggaatgt aaggttctca 360

gatgaaggag gtttcacctg cttcttccga gatcattctt accaagagga ggcagcaatg 420

gaattgaaag tagaagatcc tttctactgg gtgagccctg gaggtaccct ggaagttctg 480

ttccaggggc ccatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc 540

actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat 600

gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct 660

tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct 720

gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa 780

ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa 840

actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt 900

ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat 960

gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta 1020

gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc 1080

aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct 1140

ccaaaatcgg at 1152

<210> 105

<211> 384

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

hMOG-GST including signal sequence

<400> 105

Met Ala Ser Leu Ser Arg Pro Ser Leu Pro Ser Cys Leu Cys Ser Phe

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Val Ser Ser Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Arg His Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Val Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Pro Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Lys Asp Gln Asp Gly Asp Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Asp Ala Ile Gly Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Arg Asn Val Arg Phe Ser Asp Glu Gly Gly Phe Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Ser Pro Gly Gly Thr Leu Glu Val Leu

145 150 155 160

Phe Gln Gly Pro Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly

165 170 175

Leu Val Gln Pro Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr

180 185 190

Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys

195 200 205

Lys Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp

210 215 220

Gly Asp Val Lys Leu Thr Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala

225 230 235 240

Asp Lys His Asn Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile

245 250 255

Ser Met Leu Glu Gly Ala Val Leu Asp Ile Arg Tyr Gly Val Ser Arg

260 265 270

Ile Ala Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser

275 280 285

Lys Leu Pro Glu Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys

290 295 300

Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr

305 310 315 320

Asp Ala Leu Asp Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala

325 330 335

Phe Pro Lys Leu Val Cys Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln

340 345 350

Ile Asp Lys Tyr Leu Lys Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln

355 360 365

Gly Trp Gln Ala Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp

370 375 380

<210> 106

<211> 1152

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of mMOG-GST

including signal sequence

<400> 106

atggcctgtt tgtggagctt ctttggccc agctgcttcc tctcccttct cctcctcctt 60

ctcctccagt tgtcatgcag ctatgcagga caattcagag tgataggacc agggtatccc 120

atccgggctt tagttgggga tgaagcagag ctgccgtgcc gcatctctcc tgggaaaaat 180

gccacgggca tggaggtggg ttggtaccgt tctcccttct caagagtggt tcacctctac 240

cgaaatggca aggaccaaga tgcagagcaa gcacctgaat accggggacg cacagagctt 300

ctgaaagaga ctatcagtga gggaaaggtt acccttagga ttcagaacgt gagattctca 360

gatgaaggag gctacacctg cttcttcaga gaccactctt accaagaaga ggcagcaatg 420

gagttgaaag tggaagatcc cttctattgg gtcaaccccg gtggtaccct ggaagttctg 480

ttccaggggc ccatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc 540

actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat 600

gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct 660

tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct 720

gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa 780

ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa 840

actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt 900

ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat 960

gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta 1020

gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc 1080

aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct 1140

ccaaaatcgg at 1152

<210> 107

<211> 384

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

mMOG-GST including signal sequence

<400> 107

Met Ala Cys Leu Trp Ser Phe Ser Trp Pro Ser Cys Phe Leu Ser Leu

1 5 10 15

Leu Leu Leu Leu Leu Leu Gln Leu Ser Cys Ser Tyr Ala Gly Gln Phe

20 25 30

Arg Val Ile Gly Pro Gly Tyr Pro Ile Arg Ala Leu Val Gly Asp Glu

35 40 45

Ala Glu Leu Pro Cys Arg Ile Ser Pro Gly Lys Asn Ala Thr Gly Met

50 55 60

Glu Val Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu Tyr

65 70 75 80

Arg Asn Gly Lys Asp Gln Asp Ala Glu Gln Ala Pro Glu Tyr Arg Gly

85 90 95

Arg Thr Glu Leu Leu Lys Glu Thr Ile Ser Glu Gly Lys Val Thr Leu

100 105 110

Arg Ile Gln Asn Val Arg Phe Ser Asp Glu Gly Gly Tyr Thr Cys Phe

115 120 125

Phe Arg Asp His Ser Tyr Gln Glu Ala Ala Ala Met Glu Leu Lys Val

130 135 140

Glu Asp Pro Phe Tyr Trp Val Asn Pro Gly Gly Thr Leu Glu Val Leu

145 150 155 160

Phe Gln Gly Pro Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly

165 170 175

Leu Val Gln Pro Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr

180 185 190

Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys

195 200 205

Lys Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp

210 215 220

Gly Asp Val Lys Leu Thr Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala

225 230 235 240

Asp Lys His Asn Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile

245 250 255

Ser Met Leu Glu Gly Ala Val Leu Asp Ile Arg Tyr Gly Val Ser Arg

260 265 270

Ile Ala Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser

275 280 285

Lys Leu Pro Glu Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys

290 295 300

Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr

305 310 315 320

Asp Ala Leu Asp Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala

325 330 335

Phe Pro Lys Leu Val Cys Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln

340 345 350

Ile Asp Lys Tyr Leu Lys Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln

355 360 365

Gly Trp Gln Ala Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp

370 375 380

<210> 108

<211> 1371

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K / S354C / T366W) -FLAG

tag excluding signal sequence

<400> 108

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatgcca ggaggagatg 1080

accaagaacc aggtcagcct gtggtgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtaaagac tacaaggacg acgatgacaa g 1371

<210> 109

<211> 457

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K / S354C / T366W)

-FLAG tag excluding signal sequence

<400> 109

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Cys Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Lys Asp Tyr Lys Asp Asp Asp Asp Lys

450 455

<210> 110

<211> 1380

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-MOG01-hLG4PE (R409K / Y349C / T366S /

L368A / Y407V) -His tag excluding signal sequence

<400> 110

caggtacagc tgcagcagtc aggcgcagga ttattgaagc cttcggagac cctttccctc 60

acctgcgctg tgtctggtgg gtccttcagt ggttatact ggacctggat ccgccagcgc 120

ccagggaagg ggctggagtg gattggagaa atcaatcatc gtggaagcac cgattacaac 180

ccgtccctca agagtcgagt caccatgtca atagacacgt ccaagagcca gttctccctg 240

aatttgaaat ctgtgaccgc cgcggacacg gctgtgtatt actgtgcgag agccgcctgg 300

gggtcttgtt atgatgggac ctgctacccc gctgaatact tccaatactg gggccaggga 360

accctggtca ccgtctcctc agctagcacc aaggggccat ccgtcttccc cctggcgccc 420

tgctccagga gcacctccga gagcacagcc gccctgggct gcctggtcaa ggactacttc 480

cccgaaccgg tgacggtgtc gtggaactca ggcgccctga ccagcggcgt gcacaccttc 540

ccggctgtcc tacagtcctc aggactctac tccctcagca gcgtggtgac cgtgccctcc 600

agcagcttgg gcacgaagac ctacacctgc aacgtagatc acaagcccag caacaccaag 660

gtggacaaga gagttgagtc caaatatggt cccccatgcc caccatgccc agcacctgag 720

ttcgaggggg gaccatcagt cttcctgttc cccccaaaac ccaaggacac tctcatgatc 780

tcccggaccc ctgaggtcac gtgcgtggtg gtggacgtga gccaggaaga ccccgaggtc 840

cagttcaact ggtacgtgga tggcgtggag gtgcataatg ccaagacaaa gccgcgggag 900

gagcagttca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg 960

ctgaacggca aggagtacaa gtgcaaggtc tccaacaaag gcctcccgtc ctccatcgag 1020

aaaaccatct ccaaagccaa agggcagccc cgagagccac aggtgtgcac cctgccccca 1080

tcccaggagg agatgaccaa gaaccaggtc agcctgtcct gcgcggtcaa aggcttctac 1140

cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc 1200

acgcctcccg tgctggactc cgacggctcc ttcttcctcg tcagcaagct aaccgtggac 1260

aagagcaggt ggcaggaggg gaatgtcttc tcatgctccg tgatgcatga ggctctgcac 1320

aaccactaca cacagaagag cctctccctg tctctgggta aacacccacca ccaccaccac 1380

<210> 111

<211> 460

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-MOG01-hLG4PE (R409K / Y349C / T366S

/ L368A / Y407V) -His tag excluding signal sequence

<400> 111

Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu

65 70 75 80

Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu

100 105 110

Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

115 120 125

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser

130 135 140

Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

145 150 155 160

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

165 170 175

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

180 185 190

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr

195 200 205

Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg

210 215 220

Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu

225 230 235 240

Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

245 250 255

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

260 265 270

Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly

275 280 285

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn

290 295 300

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

305 310 315 320

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro

325 330 335

Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

340 345 350

Pro Gln Val Cys Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn

355 360 365

Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile

370 375 380

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

385 390 395 400

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys

405 410 415

Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys

420 425 430

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

435 440 445

Ser Leu Ser Leu Gly Lys His His His His

450 455 460

<210> 112

<211> 2043

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) -linker-MOG01 VL-CL

excluding signal sequence

<400> 112

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtaaagga ggaggagggt ccggaggagg agggtccgga 1380

ggaggagggt cccagtctgc cctgactcag cctgcctccg tgtctgggtc tcctggacag 1440

tcgatcacca tctcctgcac tggaaccagc cgtgacgttg gtggttataa ctatgtctcc 1500

tggtaccaac aacacccagg caaagccccc aaactcatga tttatgatgt caataatcgg 1560

ccctcagggg tttctaatcg gttctctggc tccaagtctg gcaacacggc ctccctgacc 1620

atctctgggc tccaggctga ggacgaggct gattatttct gcagctcata tacaagcagt 1680

agcacccctg tggtattcgg cggtgggacc aagctgaccg tcctaggtca gcccaaggcc 1740

gccccctcgg tcactctgtt cccgccctcc tctgaggagc ttcaagccaa caaggccaca 1800

ctggtgtgtc tcataagtga cttctacccg ggagccgtga cagtggcctg gaaggcagat 1860

agcagccccg tcaaggcggg agtggagacc accacaccct ccaaacaaag caacaacaag 1920

tacgcggcca gcagctacct gagcctgacg cctgagcagt ggaagtccca cagaagctac 1980

agctgccagg tcacgcatga aggagcacc gtggagaaga cagtggcccc tacagaatgt 2040

tca 2043

<210> 113

<211> 681

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) -linker-MOG01

VL-CL excluding signal sequence

<400> 113

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

465 470 475 480

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr

485 490 495

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

500 505 510

Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

515 520 525

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

530 535 540

Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser

545 550 555 560

Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

565 570 575

Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu

580 585 590

Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe

595 600 605

Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val

610 615 620

Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys

625 630 635 640

Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser

645 650 655

His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu

660 665 670

Lys Thr Val Ala Pro Thr Glu Cys Ser

675 680

<210> 114

<211> 675

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-MOG01 VH-CH excluding signal sequence

<400> 114

caggtacagc tgcagcagtc aggcgcagga ttattgaagc cttcggagac cctttccctc 60

acctgcgctg tgtctggtgg gtccttcagt ggttatact ggacctggat ccgccagcgc 120

ccagggaagg ggctggagtg gattggagaa atcaatcatc gtggaagcac cgattacaac 180

ccgtccctca agagtcgagt caccatgtca atagacacgt ccaagagcca gttctccctg 240

aatttgaaat ctgtgaccgc cgcggacacg gctgtgtatt actgtgcgag agccgcctgg 300

gggtcttgtt atgatgggac ctgctacccc gctgaatact tccaatactg gggccaggga 360

accctggtca ccgtctcctc agctagcacc aaggggccat ccgtcttccc cctggcgccc 420

tgctccagga gcacctccga gagcacagcc gccctgggct gcctggtcaa ggactacttc 480

cccgaaccgg tgacggtgtc gtggaactca ggcgccctga ccagcggcgt gcacaccttc 540

ccggctgtcc tacagtcctc aggactctac tccctcagca gcgtggtgac cgtgccctcc 600

agcagcttgg gcacgaagac ctacacctgc aacgtagatc acaagcccag caacaccaag 660

gtggacaaga gagt 675

<210> 115

<211> 225

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

antibody sequence of pCI-MOG01 VH-CH excluding signal sequence

<400> 115

Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu

65 70 75 80

Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu

100 105 110

Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

115 120 125

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser

130 135 140

Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

145 150 155 160

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

165 170 175

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

180 185 190

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr

195 200 205

Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg

210 215 220

Val

225

<210> 116

<211> 2181

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-AVM-hLG4PE (R409K / S354C / T366W) -linker-MOG01

scFv-FLAG tag excluding signal sequence

<400> 116

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatgcca ggaggagatg 1080

accaagaacc aggtcagcct gtggtgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gagggtccca ggtacagctg cagcagtcag gcgcaggatt attgaagcct 1440

tcggagaccc tttccctcac ctgcgctgtg tctggtgggt ccttcagtgg ttaactactgg 1500

acctggatcc gccagcgccc agggaagggg ctggagtgga ttggagaaat caatcatcgt 1560

ggaagcaccg attacaaccc gtccctcaag agtcgagtca ccatgtcaat agacacgtcc 1620

aagagccagt tctccctgaa tttgaaatct gtgaccgccg cggacacggc tgtgtattac 1680

tgtgcgagag ccgcctgggg gtcttgttat gatgggacct gctaccccgc tgaatacttc 1740

caatactggg gccagggaac cctggtcacc gtctcctcag gaggcggtgg cagcggtggg 1800

cgcgcctcgg gcggaggtgg ttcacagtct gccctgactc agcctgcctc cgtgtctggg 1860

tctcctggac agtcgatcac catctcctgc actggaacca gccgtgacgt tggtggttat 1920

aactatgtct cctggtacca acaacaccca ggcaaagccc ccaaactcat gattttatgat 1980

gtcaataatc ggccctcagg ggtttctaat cggttctctg gctccaagtc tggcaacacg 2040

gcctccctga ccatctctgg gctccaggct gaggacgagg ctgattattt ctgcagctca 2100

tatacaagca gtagcacccc tgtggtattc ggcggtggga ccaagctgac cgtcctagac 2160

tacaaggacg acgatgacaa g 2181

<210> 117

<211> 727

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K / S354C / T366W)

-linker-MOG01 scFv-FLAG tag excluding signal sequence

<400> 117

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Cys Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro

465 470 475 480

Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser

485 490 495

Gly Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu

500 505 510

Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser

515 520 525

Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe

530 535 540

Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

545 550 555 560

Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro

565 570 575

Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

580 585 590

Ser Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser

595 600 605

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

610 615 620

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr

625 630 635 640

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

645 650 655

Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

660 665 670

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

675 680 685

Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser

690 695 700

Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Asp

705 710 715 720

Tyr Lys Asp Asp Asp Asp Lys

725

<210> 118

<211> 1365

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K / Y349C / T366S /

L368A / Y407V) -His tag excluding signal sequence

<400> 118

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tgcaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gtcctgcgcg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctcgtcagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtaaacac caccaccacc accac 1365

<210> 119

<211> 455

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K / Y349C / T366S /

L368A / Y407V) -His tag excluding signal sequence

<400> 119

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser

355 360 365

Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Lys His His His

450 455

<210> 120

<211> 2151

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv2

excluding signal sequence

<400> 120

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtaaagga ggaggagggt ccggaggagg agggtccgga 1380

ggaggagggt cccaggtaca gctgcagcag tcaggcgcag gattattgaa gccttcggag 1440

accctttccc tcacctgcgc tgtgtctggt gggtccttca gtggttacta ctggacctgg 1500

atccgccagc gcccagggaa ggggctggag tggattggag aaatcaatca tcgtggaagc 1560

accgattaca accgtccct caagagtcga gtcaccatgt caatagacac gtccaagagc 1620

cagttctccc tgaatttgaa atctgtgacc gccgcggaca cggctgtgta ttatgtgcg 1680

agagccgcct gggggtcttg ttatgatggg acctgctacc ccgctgaata cttccaatac 1740

tggggccagg gaaccctggt caccgtctcc tcaggaggcg gtggcagcgg tgggcgcgcc 1800

tcgggcggag gtggttcaca gtctgccctg actcagcctg cctccgtgtc tgggtctcct 1860

ggacagtcga tcaccatctc ctgcactgga accagccgtg acgttggtgg ttataactat 1920

gtctcctggt acccaacaaca cccaggcaaa gcccccaaac tcatgattta tgatgtcaat 1980

aatcggccct caggggtttc taatcggttc tctggctcca agtctggcaa cacggcctcc 2040

ctgaccatct ctgggctcca ggctgaggac gaggctgatt atttctgcag ctcatataca 2100

agcagtagca cccctgtggt attcggcggt gggaccaagc tgaccgtcct a 2151

<210> 121

<211> 717

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 2

excluding signal sequence

<400> 121

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu

465 470 475 480

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr

485 490 495

Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile

500 505 510

Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys

515 520 525

Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu

530 535 540

Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

545 550 555 560

Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu

565 570 575

Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Gly

580 585 590

Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser Gln Ser

595 600 605

Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser Ile

610 615 620

Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn Tyr

625 630 635 640

Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile

645 650 655

Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser Gly

660 665 670

Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala

675 680 685

Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser Ser Ser Thr

690 695 700

Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

705 710 715

<210> 122

<211> 2157

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv3 excluding signal

sequence

<400> 122

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gaggtccca gtctgccctg actcagcctg cctccgtgtc tgggtctcct 1440

ggacagtcga tcaccatctc ctgcactgga accagccgtg acgttggtgg ttataactat 1500

gtctcctggt acccaacaaca cccaggcaaa gcccccaaac tcatgattta tgatgtcaat 1560

aatcggccct caggggtttc taatcggttc tctggctcca agtctggcaa cacggcctcc 1620

ctgaccatct ctgggctcca ggctgaggac gaggctgatt atttctgcag ctcatataca 1680

agcagtagca cccctgtggt attcggcggt gggaccaagc tgaccgtcct aggaggcggt 1740

ggcagcggtg ggcgcgcctc gggcggaggt ggttcacagg tacagctgca gcagtcaggc 1800

gcaggattat tgaagccttc ggagaccctt tccctcacct gcgctgtgtc tggtgggtcc 1860

ttcagtggtt actactggac ctggatccgc cagcgcccag ggaaggggct ggagtggatt 1920

ggagaaatca atcatcgtgg aagcaccgat tacaacccgt ccctcaagag tcgagtcacc 1980

atgtcaatag acacgtccaa gagccagttc tccctgaatt tgaaatctgt gaccgccgcg 2040

gacacggctg tgtattactg tgcgagagcc gcctgggggt cttgttatga tgggacctgc 2100

taccccgctg aatacttcca atactggggc cagggaaccc tggtcaccgt ctcctca 2157

<210> 123

<211> 719

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 3

excluding signal sequence

<400> 123

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro

465 470 475 480

Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly

485 490 495

Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro

500 505 510

Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn

515 520 525

Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser

530 535 540

Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr

545 550 555 560

Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val

565 570 575

Leu Gly Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser

580 585 590

Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu

595 600 605

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr

610 615 620

Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile

625 630 635 640

Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys

645 650 655

Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu

660 665 670

Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

675 680 685

Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu

690 695 700

Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

705 710 715

<210> 124

<211> 2175

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 4

excluding signal sequence

<400> 124

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga ggagggtccg gaggaggagg gtccggtgga 1380

ggtgggtccc aggtacagct gcagcagtca ggcgcaggat tattgaagcc ttcggagacc 1440

ctttccctca cctgcgctgt gtctggtggg tccttcagtg gttaactactg gacctggatc 1500

cgccagcgcc cagggaaggg gctggagtgg attggagaaa tcaatcatcg tggaagcacc 1560

gattacaacc cgtccctcaa gagtcgagtc accatgtcaa tagacacgtc caagagccag 1620

ttctccctga atttgaaatc tgtgaccgcc gcggacacgg ctgtgtatta ctgtgcgaga 1680

gccgcctggg ggtcttgtta tgatgggacc tgctaccccg ctgaatactt ccaatactgg 1740

ggccagggaa ccctggtcac cgtctcctca gctagcaccg gaggcggtgg cagcggagga 1800

ggagggtccg gtgggggcgg ctcgggcgga ggtggttcac agtctgcct gactcagcct 1860

gcctccgtgt ctgggtctcc tggacagtcg atcaccatct cctgcactgg aaccagccgt 1920

gacgttggtg gttataacta tgtctcctgg taccaacaac acccaggcaa agcccccaaa 1980

ctcatgattt atgatgtcaa taatcggccc tcaggggttt ctaatcggtt ctctggctcc 2040

aagtctggca acacggcctc cctgaccatc tctgggctcc aggctgagga cgaggctgat 2100

tatttctgca gctcatatac aagcagtagc acccctgtgg tattcggcgg tgggaccaag 2160

ctgaccgtcc taggt 2175

<210> 125

<211> 725

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 4

excluding signal sequence

<400> 125

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

450 455 460

Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr

465 470 475 480

Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr Tyr

485 490 495

Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile Gly

500 505 510

Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys Ser

515 520 525

Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu Asn

530 535 540

Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg

545 550 555 560

Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu Tyr

565 570 575

Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser

580 585 590

Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser

595 600 605

Gly Gly Gly Gly Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser

610 615 620

Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg

625 630 635 640

Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly

645 650 655

Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly

660 665 670

Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu

675 680 685

Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser

690 695 700

Ser Tyr Thr Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys

705 710 715 720

Leu Thr Val Leu Gly

725

<210> 126

<211> 2175

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 5 excluding

signal sequence

<400> 126

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga ggagggtccg gaggaggagg gtccggtgga 1380

ggtgggtccc agtctgccct gactcagcct gcctccgtgt ctgggtctcc tggacagtcg 1440

atcaccatct cctgcactgg aaccagccgt gacgttggtg gttataacta tgtctcctgg 1500

taccaacaac acccaggcaa agcccccaaa ctcatgattt atgatgtcaa taatcggccc 1560

tcaggggttt ctaatcggtt ctctggctcc aagtctggca acacggcctc cctgaccatc 1620

tctgggctcc aggctgagga cgaggctgat tatttctgca gctcatatac aagcagtagc 1680

acccctgtgg tattcggcgg tgggaccaag ctgaccgtcc taggtggagg cggtggcagc 1740

ggaggaggag ggtccggtgg gggcggctcg ggcggaggtg gttcacaggt acagctgcag 1800

cagtcaggcg caggattatt gaagccttcg gagaccctt ccctcacctg cgctgtgtct 1860

ggtgggtcct tcagtggtta ctactggacc tggatccgcc agcgcccagg gaaggggctg 1920

gagtggattg gagaaatcaa tcatcgtgga agcaccgatt acaacccgtc cctcaagagt 1980

cgagtcacca tgtcaataga cacgtccaag agccagttct ccctgaattt gaaatctgtg 2040

accgccgcgg acacggctgt gtattactgt gcgagagccg cctgggggtc ttgttatgat 2100

gggacctgct accccgctga atacttccaa tactggggcc agggaaccct ggtcaccgtc 2160

tcctcagcta gcacc 2175

<210> 127

<211> 725

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv5

excluding signal sequence

<400> 127

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

450 455 460

Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser

465 470 475 480

Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn

485 490 495

Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met

500 505 510

Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser

515 520 525

Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln

530 535 540

Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser Ser Ser

545 550 555 560

Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly

565 570 575

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys

595 600 605

Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe

610 615 620

Ser Gly Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu

625 630 635 640

Glu Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro

645 650 655

Ser Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln

660 665 670

Phe Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Ala Asp Thr Ala Val Tyr

675 680 685

Tyr Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr

690 695 700

Pro Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

705 710 715 720

Ser Ser Ala Ser Thr

725

<210> 128

<211> 2148

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv6

excluding signal sequence

<400> 128

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga ggagggtccg gaggaggagg gtccggtgga 1380

ggtgggtccc agtctgccct gactcagcct gcctccgtgt ctgggtctcc tggacagtcg 1440

atcaccatct cctgcactgg aaccagccgt gacgttggtg gttataacta tgtctcctgg 1500

taccaacaac acccaggcaa agcccccaaa ctcatgattt atgatgtcaa taatcggccc 1560

tcaggggttt ctaatcggtt ctctggctcc aagtctggca acacggcctc cctgaccatc 1620

tctgggctcc aggctgagga cgaggctgat tatttctgca gctcatatac aagcagtagc 1680

acccctgtgg tattcggcgg tgggaccaag ctgaccgtcc taggaggcgg tggcagcggt 1740

gggcgcgcct cgggcggagg tggttcacag gtacagctgc agcagtcagg cgcaggatta 1800

ttgaagcctt cggagaccct ttccctcacc tgcgctgtgt ctggtgggtc cttcagtggt 1860

tactactgga cctggatccg ccagcgccca gggaaggggc tggagtggat tggagaaatc 1920

aatcatcgtg gaagcaccga ttacaacccg tccctcaaga gtcgagtcac catgtcaata 1980

gacacgtcca agagccagtt ctccctgaat ttgaaatctg tgaccgccgc ggacacggct 2040

gtgtattact gtgcgagagc cgcctggggg tcttgttatg atgggacctg ctaccccgct 2100

gaatacttcc aatactgggg ccagggaacc ctggtcaccg tctcctca 2148

<210> 129

<211> 716

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 6

excluding signal sequence

<400> 129

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

450 455 460

Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser

465 470 475 480

Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn

485 490 495

Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met

500 505 510

Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser

515 520 525

Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln

530 535 540

Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser Ser Ser

545 550 555 560

Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly

565 570 575

Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser Gln Val Gln

580 585 590

Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu Ser

595 600 605

Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr Tyr Tyr Trp Thr

610 615 620

Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile

625 630 635 640

Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys Ser Arg Val

645 650 655

Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu Asn Leu Lys

660 665 670

Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Ala

675 680 685

Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu Tyr Phe Gln

690 695 700

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

705 710 715

<210> 130

<211> 2172

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv7

excluding signal sequence

<400> 130

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gaggtccca gtctgccctg actcagcctg cctccgtgtc tgggtctcct 1440

ggacagtcga tcaccatctc ctgcactgga accagccgtg acgttggtgg ttataactat 1500

gtctcctggt acccaacaaca cccaggcaaa gcccccaaac tcatgattta tgatgtcaat 1560

aatcggccct caggggtttc taatcggttc tctggctcca agtctggcaa cacggcctcc 1620

ctgaccatct ctgggctcca ggctgaggac gaggctgatt atttctgcag ctcatataca 1680

agcagtagca cccctgtggt attcggcggt gggaccaagc tgaccgtcct aggaggcggt 1740

ggcagcggag gaggagggtc cggtgggggc ggctcgggcg gaggtggttc acaggtacag 1800

ctgcagcagt caggcgcagg attattgaag ccttcggaga ccctttccct cacctgcgct 1860

gtgtctggtg ggtccttcag tggttatactac tggacctgga tccgccagcg cccagggaag 1920

gggctggagt ggattggaga aatcaatcat cgtggaagca ccgattacaa cccgtccctc 1980

aagagtcgag tcaccatgtc aatagacacg tccaagagcc agttctccct gaatttgaaa 2040

tctgtgaccg ccgcggacac ggctgtgtat tactgtgcga gagccgcctg ggggtcttgt 2100

tatgatggga cctgctaccc cgctgaatac ttccaatact ggggccaggg aaccctggtc 2160

accgtctcct ca 2172

<210> 131

<211> 724

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv7

excluding signal sequence

<400> 131

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro

465 470 475 480

Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly

485 490 495

Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro

500 505 510

Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn

515 520 525

Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser

530 535 540

Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr

545 550 555 560

Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val

565 570 575

Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

580 585 590

Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu

595 600 605

Leu Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly

610 615 620

Ser Phe Ser Gly Tyr Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys

625 630 635 640

Gly Leu Glu Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr

645 650 655

Asn Pro Ser Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys

660 665 670

Ser Gln Phe Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala

675 680 685

Val Tyr Tyr Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr

690 695 700

Cys Tyr Pro Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val

705 710 715 720

Thr Val Ser Ser

<210> 132

<211> 2169

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 8

excluding signal sequence

<400> 132

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gaggtccca gtctgccctg actcagcctg cctccgtgtc tgggtctcct 1440

ggacagtcga tcaccatctc ctgcactgga accagccgtg acgttggtgg ttataactat 1500

gtctcctggt acccaacaaca cccaggcaaa gcccccaaac tcatgattta tgatgtcaat 1560

aatcggccct caggggtttc taatcggttc tctggctcca agtctggcaa cacggcctcc 1620

ctgaccatct ctgggctcca ggctgaggac gaggctgatt atttctgcag ctcatataca 1680

agcagtagca cccctgtggt attcggcggt gggaccaagc tgaccgtcct aggtggaggc 1740

ggtggcagcg gtgggcgcgc ctcgggcgga ggtggttcac aggtacagct gcagcagtca 1800

ggcgcaggat tattgaagcc ttcggagacc ctttccctca cctgcgctgt gtctggtggg 1860

tccttcagtg gttaactactg gacctggatc cgccagcgcc cagggaaggg gctggagtgg 1920

attggagaaa tcaatcatcg tggaagcacc gattacaacc cgtccctcaa gagtcgagtc 1980

accatgtcaa tagacacgtc caagagccag ttctccctga atttgaaatc tgtgaccgcc 2040

gcggacacgg ctgtgtatta ctgtgcgaga gccgcctggg ggtcttgtta tgatgggacc 2100

tgctaccccg ctgaatactt ccaatactgg ggccagggaa ccctggtcac cgtctcctca 2160

gctagcacc 2169

<210> 133

<211> 723

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 8

excluding signal sequence

<400> 133

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro

465 470 475 480

Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly

485 490 495

Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro

500 505 510

Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn

515 520 525

Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser

530 535 540

Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr

545 550 555 560

Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val

565 570 575

Leu Gly Gly Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly

580 585 590

Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser

595 600 605

Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly

610 615 620

Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp

625 630 635 640

Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu

645 650 655

Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser

660 665 670

Leu Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

675 680 685

Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala

690 695 700

Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

705 710 715 720

Ala Ser Thr

<210> 134

<211> 2163

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv9 excluding signal

sequence

<400> 134

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga ggagggtccg gaggaggagg gtccggtgga 1380

ggtgggtccc agtctgccct gactcagcct gcctccgtgt ctgggtctcc tggacagtcg 1440

atcaccatct cctgcactgg aaccagccgt gacgttggtg gttataacta tgtctcctgg 1500

taccaacaac acccaggcaa agcccccaaa ctcatgattt atgatgtcaa taatcggccc 1560

tcaggggttt ctaatcggtt ctctggctcc aagtctggca acacggcctc cctgaccatc 1620

tctgggctcc aggctgagga cgaggctgat tatttctgca gctcatatac aagcagtagc 1680

acccctgtgg tattcggcgg tgggaccaag ctgaccgtcc taggaggcgg tggcagcgga 1740

ggaggagggt ccggtggggg cggctcgggc ggaggtggtt cacaggtaca gctgcagcag 1800

tcaggcgcag gattattgaa gccttcggag acctttccc tcacctgcgc tgtgtctggt 1860

gggtccttca gtggttacta ctggacctgg atccgccagc gcccagggaa ggggctggag 1920

tggattggag aaatcaatca tcgtggaagc accgattaca accgtccct caagagtcga 1980

gtcaccatgt caatagacac gtccaagagc cagttctccc tgaatttgaa atctgtgacc 2040

gccgcggaca cggctgtgta ttatactgtgcg agagccgcct gggggtcttg ttatgatggg 2100

acctgctacc ccgctgaata cttccaatac tggggccagg gaaccctggt caccgtctcc 2160

tca 2163

<210> 135

<211> 721

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv9

excluding signal sequence

<400> 135

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

450 455 460

Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser

465 470 475 480

Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn

485 490 495

Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met

500 505 510

Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser

515 520 525

Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln

530 535 540

Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser Ser Ser

545 550 555 560

Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

580 585 590

Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro

595 600 605

Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser

610 615 620

Gly Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu

625 630 635 640

Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser

645 650 655

Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe

660 665 670

Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

675 680 685

Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro

690 695 700

Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

705 710 715 720

Ser

<210> 136

<211> 2160

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 10 excluding

signal sequence

<400> 136

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga ggagggtccg gaggaggagg gtccggtgga 1380

ggtgggtccc agtctgccct gactcagcct gcctccgtgt ctgggtctcc tggacagtcg 1440

atcaccatct cctgcactgg aaccagccgt gacgttggtg gttataacta tgtctcctgg 1500

taccaacaac acccaggcaa agcccccaaa ctcatgattt atgatgtcaa taatcggccc 1560

tcaggggttt ctaatcggtt ctctggctcc aagtctggca acacggcctc cctgaccatc 1620

tctgggctcc aggctgagga cgaggctgat tatttctgca gctcatatac aagcagtagc 1680

acccctgtgg tattcggcgg tgggaccaag ctgaccgtcc taggtggagg cggtggcagc 1740

ggtgggcgcg cctcgggcgg aggtggttca caggtacagc tgcagcagtc aggcgcagga 1800

ttattgaagc cttcggagac cctttccctc acctgcgctg tgtctggtgg gtccttcagt 1860

ggttaactact ggacctggat ccgccagcgc ccagggaagg ggctggagtg gattggagaa 1920

atcaatcatc gtggaagcac cgattacaac ccgtccctca agagtcgagt caccatgtca 1980

atagacacgt ccaagagcca gttctccctg aatttgaaat ctgtgaccgc cgcggacacg 2040

gctgtgtatt actgtgcgag agccgcctgg gggtcttgtt atgatgggac ctgctacccc 2100

gctgaatact tccaatactg gggccaggga accctggtca ccgtctcctc agctagcacc 2160

<210> 137

<211> 720

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 10

excluding signal sequence

<400> 137

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

450 455 460

Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser

465 470 475 480

Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn

485 490 495

Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met

500 505 510

Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser

515 520 525

Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln

530 535 540

Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr Ser Ser Ser

545 550 555 560

Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly

565 570 575

Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser Gln Val

580 585 590

Gln Leu Gln Gln Ser Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu

595 600 605

Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Phe Ser Gly Tyr Tyr Trp

610 615 620

Thr Trp Ile Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu

625 630 635 640

Ile Asn His Arg Gly Ser Thr Asp Tyr Asn Pro Ser Leu Lys Ser Arg

645 650 655

Val Thr Met Ser Ile Asp Thr Ser Lys Ser Gln Phe Ser Leu Asn Leu

660 665 670

Lys Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala

675 680 685

Ala Trp Gly Ser Cys Tyr Asp Gly Thr Cys Tyr Pro Ala Glu Tyr Phe

690 695 700

Gln Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr

705 710 715 720

<210> 138

<211> 2184

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG01 scFv11

excluding signal sequence

<400> 138

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gaggtccca gtctgccctg actcagcctg cctccgtgtc tgggtctcct 1440

ggacagtcga tcaccatctc ctgcactgga accagccgtg acgttggtgg ttataactat 1500

gtctcctggt acccaacaaca cccaggcaaa gcccccaaac tcatgattta tgatgtcaat 1560

aatcggccct caggggtttc taatcggttc tctggctcca agtctggcaa cacggcctcc 1620

ctgaccatct ctgggctcca ggctgaggac gaggctgatt atttctgcag ctcatataca 1680

agcagtagca cccctgtggt attcggcggt gggaccaagc tgaccgtcct aggtggaggc 1740

ggtggcagcg gaggaggagg gtccggtggg ggcggctcgg gcggaggtgg ttcacaggta 1800

cagctgcagc agtcaggcgc aggattattg aagccttcgg agaccctttc cctcacctgc 1860

gctgtgtctg gtgggtcctt cagtggttac tactggacct ggatccgcca gcgcccaggg 1920

aaggggctgg agtggattgg agaaatcaat catcgtggaa gcaccgatta caacccgtcc 1980

ctcaagagtc gagtcaccat gtcaatagac acgtccaaga gccagttctc cctgaatttg 2040

aaatctgtga ccgccgcgga cacggctgtg tattactgtg cgagagccgc ctgggggtct 2100

tgttatgatg ggacctgcta ccccgctgaa tacttccaat actggggcca gggaaccctg 2160

gtcaccgtct cctcagctag cacc 2184

<210> 139

<211> 728

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _MOG 01 scFv 11

excluding signal sequence

<400> 139

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro

465 470 475 480

Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Arg Asp Val Gly

485 490 495

Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro

500 505 510

Lys Leu Met Ile Tyr Asp Val Asn Asn Arg Pro Ser Gly Val Ser Asn

515 520 525

Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser

530 535 540

Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Phe Cys Ser Ser Tyr Thr

545 550 555 560

Ser Ser Ser Thr Pro Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val

565 570 575

Leu Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Gly

595 600 605

Leu Leu Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Ser Gly

610 615 620

Gly Ser Phe Ser Gly Tyr Tyr Trp Thr Trp Ile Arg Gln Arg Pro Gly

625 630 635 640

Lys Gly Leu Glu Trp Ile Gly Glu Ile Asn His Arg Gly Ser Thr Asp

645 650 655

Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Met Ser Ile Asp Thr Ser

660 665 670

Lys Ser Gln Phe Ser Leu Asn Leu Lys Ser Val Thr Ala Ala Asp Thr

675 680 685

Ala Val Tyr Tyr Cys Ala Arg Ala Ala Trp Gly Ser Cys Tyr Asp Gly

690 695 700

Thr Cys Tyr Pro Ala Glu Tyr Phe Gln Tyr Trp Gly Gln Gly Thr Leu

705 710 715 720

Val Thr Val Ser Ser Ala Ser Thr

725

<210> 140

<211> 2040

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) -linker-AVMVL-CL

excluding signal sequence

<400> 140

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtaaagga ggaggagggt ccggaggagg agggtccgga 1380

ggaggagggt cccagtttgt gctttctcag ccaaactctg tgtctacgaa tctcggaagc 1440

acagtcaaac tgtcttgcaa gcgcagcact ggtaacattg gaagcaatta tgtgagctgg 1500

taccagcagc atgagggaag atctcccacc actatgattt atagggatga taagagacca 1560

gatggagttc ctgacaggtt ctctggctcc attgacagat cttccgactc agccctcctg 1620

acaatcaata atgtgcagac tgaagatgaa gctgactact tctgtcagtc ttacagtagt 1680

ggtattaata ttttcggcgg tggaaccaag ctcactgtcc taggtcagcc caaggccgcc 1740

ccctcggtca ctctgttccc gccctcctct gaggagcttc aagccaacaa ggccacactg 1800

gtgtgtctca taagtgactt ctacccggga gccgtgacag tggcctggaa ggcagatagc 1860

agccccgtca aggcgggagt ggagaccacc acaccctcca aacaaagcaa caacaagtac 1920

gcggccagca gctacctgag cctgacgcct gagcagtgga agtcccacag aagctacagc 1980

tgccaggtca cgcatgaagg gagcaccgtg gagaagacag tggcccctac agaatgttca 2040

<210> 141

<211> 680

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) -linker-AVMVL-CL

excluding signal sequence

<400> 141

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Phe Val Leu Ser Gln Pro Asn Ser Val Ser Thr Asn Leu Gly Ser

465 470 475 480

Thr Val Lys Leu Ser Cys Lys Arg Ser Thr Gly Asn Ile Gly Ser Asn

485 490 495

Tyr Val Ser Trp Tyr Gln Gln His Glu Gly Arg Ser Pro Thr Thr Met

500 505 510

Ile Tyr Arg Asp Asp Lys Arg Pro Asp Gly Val Pro Asp Arg Phe Ser

515 520 525

Gly Ser Ile Asp Arg Ser Ser Asp Ser Ala Leu Leu Thr Ile Asn Asn

530 535 540

Val Gln Thr Glu Asp Glu Ala Asp Tyr Phe Cys Gln Ser Tyr Ser Ser

545 550 555 560

Gly Ile Asn Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

565 570 575

Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

580 585 590

Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr

595 600 605

Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys

610 615 620

Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr

625 630 635 640

Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His

645 650 655

Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys

660 665 670

Thr Val Ala Pro Thr Glu Cys Ser

675 680

<210> 142

<211> 660

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of antibody

sequence of pCI-AVMVH-CH excluding signal sequence

<400> 142

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

<210> 143

<211> 220

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

antibody sequence of pCI-AVMVH-CH excluding signal sequence

<400> 143

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val

210 215 220

<210> 144

<211> 2163

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K / S354C / T366W)

-linker-AVMscFv-FLAG tag excluding signal sequence

<400> 144

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatgcca ggaggagatg 1080

accaagaacc aggtcagcct gtggtgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gagggtccga ggtgcagctg gtggaatctg gggaggctt agtgcagcct 1440

ggaagatccc tgaaactctc ctgtgcagcc tcaggattca ctttcagtaa ctatgccatg 1500

gcttgggtcc gccgggctcc aacgaagggt ctggagtggg tcgcatccat tagtaatggt 1560

ggtggtaaca cttactatcg cgactccgtg aagggccgat tcactatctc cagagatgat 1620

gcaaaaaaca ccctatacct gcaaatggac agtctgaggt ctgaggacac ggccacttat 1680

tactgtgcaa gacacgggaa ttatatatat tatgggtcct tctttgatta ctggggccaa 1740

ggagtcatgg tcacagtctc ctcaggaggc ggtggcagcg gtgggcgcgc ctcgggcgga 1800

ggtggttcac agtttgtgct ttctcagcca aactctgtgt ctacgaatct cggaagcaca 1860

gtcaaactgt cttgcaagcg cagcactggt aacattggaa gcaattatgt gagctggtac 1920

cagcagcatg agggaagatc tcccaccact atgatttata gggatgataa gagaccat 1980

ggagttcctg acaggttctc tggctccatt gacagatctt ccgactcagc cctcctgaca 2040

atcaataatg tgcagactga agatgaagct gactacttct gtcagtctta cagtagtggt 2100

attaatattt tcggcggtgg aaccaagctc actgtcctag actacaagga cgacgatgac 2160

aag 2163

<210> 145

<211> 721

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K / S354C / T366W)

-linker-AVMscFv-FLAG tag excluding signal sequence

<400> 145

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Cys Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

465 470 475 480

Gly Arg Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

485 490 495

Asn Tyr Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu

500 505 510

Trp Val Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp

515 520 525

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr

530 535 540

Leu Tyr Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr

545 550 555 560

Tyr Cys Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp

565 570 575

Tyr Trp Gly Gln Gly Val Met Val Thr Val Ser Gly Gly Gly Gly

580 585 590

Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser Gln Phe Val Leu Ser

595 600 605

Gln Pro Asn Ser Val Ser Thr Asn Leu Gly Ser Thr Val Lys Leu Ser

610 615 620

Cys Lys Arg Ser Thr Gly Asn Ile Gly Ser Asn Tyr Val Ser Trp Tyr

625 630 635 640

Gln Gln His Glu Gly Arg Ser Pro Thr Thr Met Ile Tyr Arg Asp Asp

645 650 655

Lys Arg Pro Asp Gly Val Pro Asp Arg Phe Ser Gly Ser Ile Asp Arg

660 665 670

Ser Ser Asp Ser Ala Leu Leu Thr Ile Asn Asn Val Gln Thr Glu Asp

675 680 685

Glu Ala Asp Tyr Phe Cys Gln Ser Tyr Ser Ser Gly Ile Asn Ile Phe

690 695 700

Gly Gly Gly Thr Lys Leu Thr Val Leu Asp Tyr Lys Asp Asp Asp Asp

705 710 715 720

Lys

<210> 146

<211> 2139

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _AVMscFv3 excluding

signal sequence

<400> 146

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga agcggaggag gagggtccgg aggaggaggg 1380

tccggaggag gaggtccca gtttgtgctt tctcagccaa actctgtgtc tacgaatctc 1440

ggaagcacag tcaaactgtc ttgcaagcgc agcactggta acattggaag caattatgtg 1500

agctggtacc agcagcatga gggaagatct cccaccacta tgatttatag ggatgataag 1560

agaccagatg gagttcctga caggttctct ggctccattg agagatcttc cgactcagcc 1620

ctcctgacaa tcaataatgt gcagactgaa gatgaagctg actacttctg tcagtcttac 1680

agtagtggta ttaatatttt cggcggtgga accaagctca ctgtcctagg aggcggtggc 1740

agcggtgggc gcgcctcggg cggaggtggt tcagaggtgc agctggtgga atctggggga 1800

ggcttagtgc agcctggaag atccctgaaa ctctcctgtg cagcctcagg atcactttc 1860

agtaactatg ccatggcttg ggtccgccgg gctccaacga agggtctgga gtgggtcgca 1920

tccattagta atggtggtgg taacacttac tatcgcgact ccgtgaaggg ccgattcact 1980

atctccagag atgatgcaaa aaacacccta tacctgcaaa tggacagtct gaggtctgag 2040

gacacggcca cttattactg tgcaagacac gggaattata tatattatgg gtccttcttt 2100

gattactggg gccaaggagt catggtcaca gtctcctca 2139

<210> 147

<211> 713

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _AVMscFv3

excluding signal sequence

<400> 147

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gln Phe Val Leu Ser Gln Pro Asn Ser Val Ser Thr Asn Leu

465 470 475 480

Gly Ser Thr Val Lys Leu Ser Cys Lys Arg Ser Thr Gly Asn Ile Gly

485 490 495

Ser Asn Tyr Val Ser Trp Tyr Gln Gln His Glu Gly Arg Ser Pro Thr

500 505 510

Thr Met Ile Tyr Arg Asp Asp Lys Arg Pro Asp Gly Val Pro Asp Arg

515 520 525

Phe Ser Gly Ser Ile Asp Arg Ser Ser Asp Ser Ala Leu Leu Thr Ile

530 535 540

Asn Asn Val Gln Thr Glu Asp Glu Ala Asp Tyr Phe Cys Gln Ser Tyr

545 550 555 560

Ser Ser Gly Ile Asn Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

565 570 575

Gly Gly Gly Gly Ser Gly Gly Arg Ala Ser Gly Gly Gly Gly Ser Glu

580 585 590

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser

595 600 605

Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr Ala

610 615 620

Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val Ala

625 630 635 640

Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val Lys

645 650 655

Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr Leu

660 665 670

Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

675 680 685

Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp Gly

690 695 700

Gln Gly Val Met Val Thr Val Ser Ser

705 710

<210> 148

<211> 2157

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of the

antibody sequence of pCI-AVM-hLG4PE (R409K) _AVMscFv5 excluding

signal sequence

<400> 148

gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60

tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120

ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180

cgcgactccg tgaagggccg atcactatc tccagagatg atgcaaaaaa caccctatac 240

ctgcaaatgg agagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300

aattatatat atttgggtc cttctttgat tactggggcc aaggagtcat ggtcacagtc 360

tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420

tccgagagca cagccgcct gggctgcctg gtcaaggact acttccccga accggtgacg 480

gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540

tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600

aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660

gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720

tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780

gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840

gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900

acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaaggag 960

tacaagtgca aggtctccaa caaaggcctc ccgtcctcca tcgagaaaac catctccaaa 1020

gccaaagggc agccccgaga gccacaggtg tacaccctgc ccccatccca ggaggagatg 1080

accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctaccccag cgacatcgcc 1140

gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200

gactccgacg gctccttctt cctctacagc aagctaaccg tggacaagag caggtggcag 1260

gaggggaatg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacacag 1320

aagagcctct ccctgtctct gggtggagga ggagggtccg gaggaggagg gtccggtgga 1380

ggtgggtccc agtttgtgct ttctcagcca aactctgtgt ctacgaatct cggaagcaca 1440

gtcaaactgt cttgcaagcg cagcactggt aacattggaa gcaattatgt gagctggtac 1500

cagcagcatg agggaagatc tcccaccact atgatttata gggatgataa gagaccat 1560

ggagttcctg acaggttctc tggctccatt gacagatctt ccgactcagc cctcctgaca 1620

atcaataatg tgcagactga agatgaagct gactacttct gtcagtctta cagtagtggt 1680

attaatattt tcggcggtgg aaccaagctc actgtcctag gtggaggcgg tggcagcgga 1740

ggaggagggt ccggtggggg cggctcgggc ggaggtggtt cagaggtgca gctggtggaa 1800

tctggggggag gcttagtgca gcctggaaga tccctgaaac tctcctgtgc agcctcagga 1860

ttcactttca gtaactatgc catggcttgg gtccgccggg ctccaacgaa gggtctggag 1920

tgggtcgcat ccattagtaa tggtggtggt aacacttact atcgcgactc cgtgaagggc 1980

cgattcacta tctccagaga tgatgcaaaa aacaccctat acctgcaaat ggacagtctg 2040

aggtctgagg acacggccac ttattactgt gcaagacacg ggaattatat atattatggg 2100

tccttctttg attactgggg ccaaggagtc atggtcacag tctcctcagc tagcacc 2157

<210> 149

<211> 719

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of

the antibody sequence of pCI-AVM-hLG4PE (R409K) _AVMscFv5

excluding signal sequence

<400> 149

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Tyr Thr Cys Asn Val Asp

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp

260 265 270

Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln

450 455 460

Phe Val Leu Ser Gln Pro Asn Ser Val Ser Thr Asn Leu Gly Ser Thr

465 470 475 480

Val Lys Leu Ser Cys Lys Arg Ser Thr Gly Asn Ile Gly Ser Asn Tyr

485 490 495

Val Ser Trp Tyr Gln Gln His Glu Gly Arg Ser Pro Thr Thr Met Ile

500 505 510

Tyr Arg Asp Asp Lys Arg Pro Asp Gly Val Pro Asp Arg Phe Ser Gly

515 520 525

Ser Ile Asp Arg Ser Ser Asp Ser Ala Leu Leu Thr Ile Asn Asn Val

530 535 540

Gln Thr Glu Asp Glu Ala Asp Tyr Phe Cys Gln Ser Tyr Ser Ser Gly

545 550 555 560

Ile Asn Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Gly Gly Gly

580 585 590

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

595 600 605

Gly Arg Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

610 615 620

Asn Tyr Ala Met Ala Trp Val Arg Arg Ala Pro Thr Lys Gly Leu Glu

625 630 635 640

Trp Val Ala Ser Ile Ser Asn Gly Gly Gly Asn Thr Tyr Tyr Arg Asp

645 650 655

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Thr

660 665 670

Leu Tyr Leu Gln Met Asp Ser Leu Arg Ser Glu Asp Thr Ala Thr Tyr

675 680 685

Tyr Cys Ala Arg His Gly Asn Tyr Ile Tyr Tyr Gly Ser Phe Phe Asp

690 695 700

Tyr Trp Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr

705 710 715

<210> 150

<211> 1887

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence of Acid

Sphingomyelinase (ASM)

<400> 150

atgccccgct acggagcgtc actccgccag agctgcccca ggtccggccg ggagcaggga 60

caagacggga ccgccggagc ccccggactc ctttggatgg gcctggtgct ggcgctggcg 120

ctggcgctgg cgctggctct gtctgactct cgggttctct gggctccggc agaggctcac 180

cctctttctc cccaaggcca tcctgccagg ttacatcgca tagtgccccg gctccgagat 240

gtctttgggt gggggaacct cacctgccca atctgcaaag gtctattcac cgccatcaac 300

ctcgggctga agaaggaacc caatgtggct cgcgtgggct ccgtggccat caagctgtgc 360

aatctgctga agatagcacc acctgccgtg tgccaatcca ttgtccacct ctttgaggat 420

gacatggtgg aggtgtggag acgctcagtg ctgagcccat ctgaggcctg tggcctgctc 480

ctgggctcca cctgtgggca ctgggacatt ttctcatctt ggaacatctc tttgcctact 540

gtgccgaagc cgccccccaa acccctagc cccccagccc caggtgcccc tgtcagccgc 600

atcctcttcc tcactgacct gcactggggat catgactacc tggagggcac ggaccctgac 660

tgtgcagacc cactgtgctg ccgccggggt tctggcctgc cgcccgcatc ccggccaggt 720

gccggatact ggggcgaata cagcaagtgt gacctgcccc tgaggaccct ggagagcctg 780

ttgagtgggc tgggcccagc cggccctttt gatatggtgt actggacagg agacatcccc 840

gcacatgatg tctggcacca gactcgtcag gaccaactgc gggccctgac caccgtcaca 900

gcacttgtga ggaagttcct ggggccagtg ccagtgtacc ctgctgtggg taaccatgaa 960

agcatacctg tcaatagctt ccctcccccc ttcattgagg gcaaccactc ctcccgctgg 1020

ctctatgaag cgatggccaa ggcttgggag ccctggctgc ctgccgaagc cctgcgcacc 1080

ctcagaattg gggggttcta tgctctttcc ccataccccg gtctccgcct catctctctc 1140

aatatgaatt tttgttcccg tgagaacttc tggctcttga tcaactccac ggatcccgca 1200

ggacagctcc agtggctggt gggggagctt caggctgctg aggatcgagg agacaaagtg 1260

catataattg gccacattcc cccagggcac tgtctgaaga gctggagctg gaattattac 1320

cgaattgtag ccaggtatga gaacaccctg gctgctcagt tctttggcca cactcatgtg 1380

gatgaatttg aggtcttcta tgatgaagag actctgagcc ggccgctggc tgtagccttc 1440

ctggcaccca gtgcaactac ctacatcggc cttaatcctg gttaccgtgt gtaccaaata 1500

gatggaaact actccaggag ctctcacgtg gtcctggacc atgagaccta catcctgaat 1560

ctgacccagg caaacatacc gggagccata ccgcactggc agcttctcta cagggctcga 1620

gaaacctatg ggctgcccaa cacactgcct accgcctggc acaacctggt atatcgcatg 1680

cggggcgaca tgcaactttt ccagaccttc tggtttctct accataaggg ccaccaccc 1740

tcggagccct gtggcacgcc ctgccgtctg gctactcttt gtgcccagct ctctgcccgt 1800

gctgacagcc ctgctctgtg ccgccacctg atgccagatg ggagcctccc agaggcccag 1860

agcctgtggc caaggccact gttttgc 1887

<210> 151

<211> 369

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence encoding

VH excluding signal sequence of MOG 301

<400> 151

caggtgcagc tggtgcagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggtta cagctttacc agctatggta tcaactgggt gcgacaggcc 120

ccaggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtta cacaaactat 180

gcacagaagc tccagggcag agtcaccatg accagagaca catccacgcg cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagagtac 300

gatattttga ctggttatattc cgatgctttt gatatctggg gccaagggac cctggtcact 360

gtctcctca 369

<210> 152

<211> 123

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 301

<400> 152

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr

20 25 30

Gly Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Tyr Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Arg Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Tyr Asp Ile Leu Thr Gly Tyr Ser Asp Ala Phe Asp Ile

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 153

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 301

<400> 153

Ser Tyr Gly Ile Asn

1 5

<210> 154

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 301

<400> 154

Trp Ile Ser Ala Tyr Asn Gly Tyr Thr Asn Tyr Ala Gln Lys Leu Gln

1 5 10 15

Gly

<210> 155

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 301

<400> 155

Glu Tyr Asp Ile Leu Thr Gly Tyr Ser Asp Ala Phe Asp Ile

1 5 10

<210> 156

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: base sequence encoding

VL excluding signal sequence of MOG 301

<400> 156

gaaatagtga tgacacagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccgct cactttcggc 300

ggagggacca aggtggaaat caaa 324

<210> 157

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> Description of the artificial sequence: amino acid sequence of VL

excluding signal sequence of MOG 301

<400> 157

Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 158

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 301

<400> 158

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 159

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 301

<400> 159

Gly Ala Ser Ser Arg Ala Thr

1 5

<210> 160

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 301

<400> 160

Gln Gln Tyr Gly Ser Ser Pro Leu Thr

1 5

<210> 161

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 303

<400> 161

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgggctgggt ccgccaggct 120

ccagggaagg ggctggaatg ggtctcagct gttagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaaggagga 300

tacgatattt tgactggtta cttctttgac tactggggcc agggaaccac ggtcactgtc 360

tcctca 366

<210> 162

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG303

<400> 162

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Val Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Gly Tyr Asp Ile Leu Thr Gly Tyr Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 163

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG303

<400> 163

Ser Tyr Ala Met Gly

1 5

<210> 164

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG303

<400> 164

Ala Val Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 165

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG303

<400> 165

Gly Gly Tyr Asp Ile Leu Thr Gly Tyr Phe Phe Asp Tyr

1 5 10

<210> 166

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 303

<400> 166

gccatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt accattcac tttcggccct 300

gggacacgac tggagattaa a 321

<210> 167

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 303

<400> 167

Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 168

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 303

<400> 168

Arg Ala Ser Gln Gly Ile Ser Ser Ala Leu Ala

1 5 10

<210> 169

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 303

<400> 169

Asp Ala Ser Ser Leu Glu Ser

1 5

<210> 170

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 303

<400> 170

Gln Gln Phe Asn Ser Tyr Pro Phe Thr

1 5

<210> 171

<211> 381

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH (excluding signal sequence) of MOG 307

<400> 171

cgggtcacct tgagggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60

acctgcacct tctctgggtt ctcactcagc actagtggag tgggtgtggg ctggatccgt 120

cagcccccag gaaaggccct ggagtggctt gcactcattt tttggggatga tgatagtcac 180

tacagcccat ctctgaagag caggctcacc atcaccaagg acacctccaa aaaccaggtg 240

gtccttacaa tgaccaacat ggaccctgtg gacacagcca catattactg tgcacggtat 300

tactttggtt cggggagtta tttccctagc tactggtact tcgatctctg gggccgtggc 360

accctggtca ctgtctcctc a 381

<210> 172

<211> 127

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 307

<400> 172

Arg Val Thr Leu Arg Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser

20 25 30

Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu

35 40 45

Trp Leu Ala Leu Ile Phe Trp Asp Asp Asp Ser His Tyr Ser Pro Ser

50 55 60

Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val

65 70 75 80

Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Ala Arg Tyr Tyr Phe Gly Ser Gly Ser Tyr Phe Pro Ser Tyr Trp

100 105 110

Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 173

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 307

<400> 173

Thr Ser Gly Val Gly Val Gly

1 5

<210> 174

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 307

<400> 174

Leu Ile Phe Trp Asp Asp Asp Ser His Tyr Ser Pro Ser Leu Lys Ser

1 5 10 15

<210> 175

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 307

<400> 175

Tyr Tyr Phe Gly Ser Gly Ser Tyr Phe Pro Ser Tyr Phe Trp Tyr Phe Asp

1 5 10 15

Leu

<210> 176

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 307

<400> 176

gaaatagtga tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccgac gttcggccaa 300

gggaccaagg tggagatcaa a 321

<210> 177

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 307

<400> 177

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 178

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 307

<400> 178

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 179

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 307

<400> 179

Asp Ala Ser Asn Arg Ala Thr

1 5

<210> 180

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 307

<400> 180

Gln Gln Arg Ser Asn Trp Pro Pro Thr

1 5

<210> 181

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 310

<400> 181

caggtgcagc tggtgcaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttagtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatgggctt taactactgg ggccagggaa ccctggtcac tgtctcctca 360

<210> 182

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 310

<400> 182

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Ser Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asn Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 183

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 310

<400> 183

Arg Tyr Ala Ile Ser

1 5

<210> 184

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 310

<400> 184

Gly Ile Ile Pro Met Phe Ser Thr Ala Asn Tyr Ala Gln Lys Phe Gln

1 5 10 15

Gly

<210> 185

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 310

<400> 185

Asp Trp Ala Val Ala Gly Met Gly Phe Asn Tyr

1 5 10

<210> 186

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 310

<400> 186

gaaattgtgt tgacacagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttatgtcag cagtatggta gctcaccgta cacttttggc 300

caggggacca aagtggatat caaa 324

<210> 187

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 310

<400> 187

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Val Asp Ile Lys

100 105

<210> 188

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR1 of MOG 310

<400> 188

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 189

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 310

<400> 189

Gly Ala Ser Ser Arg Ala Thr

1 5

<210> 190

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR3 of MOG 310

<400> 190

Gln Gln Tyr Gly Ser Ser Pro Tyr Thr

1 5

<210> 191

<211> 354

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 312

<400> 191

caggtgcagc tacagcagtg gggcgcagga ctgttgaagc cttcggagac cctgtccctc 60

acctgcgctg tctatggtgg gtccttaagt ggttatact ggagctggat ccgccagccc 120

ccagggaagg ggctggagtg gattggggat atcactcata gtggaagcac caactacaac 180

ccgtccctca agagtcgagt caccatgtca gttgacacgt ccaaaaacca gttctccctg 240

aacctgaact ctgtgaccgc cgcggacacg gctgtgtatt actgtgcgag aagggggata 300

ggagctgctg tctttgacct ctggggccag ggaaccctgg tcactgtctc ctca 354

<210> 192

<211> 118

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 312

<400> 192

Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Leu Ser Gly Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Asp Ile Thr His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Asn Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Arg Gly Ile Gly Ala Ala Val Phe Asp Leu Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 193

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 312

<400> 193

Gly Tyr Tyr Trp Ser

1 5

<210> 194

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 312

<400> 194

Asp Ile Thr His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 195

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 312

<400> 195

Arg Gly Ile Gly Ala Ala Val Phe Asp Leu

1 5 10

<210> 196

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 312

<400> 196

gaaatagtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttactgtcag cagcgtagca actggcctct cactttcggc 300

ggagggacca aggtggagat caaa 324

<210> 197

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 312

<400> 197

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro

85 90 95

Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 198

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 312

<400> 198

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 199

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 312

<400> 199

Gly Ala Ser Ser Arg Ala Thr

1 5

<210> 200

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 312

<400> 200

Gln Gln Arg Ser Asn Trp Pro Leu Thr

1 5

<210> 201

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 326

<400> 201

gaggtccagc tggtacagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttcacc agctactata tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atcaacccta gtggtggtag cacaagctac 180

gcacagaagt tccagggcag agtcaccatg accagggaca cgtccacgag cacagtctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagggtat 300

tacgatattt tgactggttc cttctttgac tactggggcc agggaaccct ggtcactgtc 360

tcctca 366

<210> 202

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 326

<400> 202

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Tyr Tyr Asp Ile Leu Thr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 203

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 326

<400> 203

Ser Tyr Tyr Met His

1 5

<210> 204

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 326

<400> 204

Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe Gln

1 5 10 15

Gly

<210> 205

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 326

<400> 205

Gly Tyr Tyr Asp Ile Leu Thr Gly Ser Phe Phe Asp Tyr

1 5 10

<210> 206

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 326

<400> 206

gacatcgtga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag tctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt accctctcac tttcggcgga 300

gggaccaaag tggatatcaa a 321

<210> 207

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 326

<400> 207

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys

100 105

<210> 208

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 326

<400> 208

Arg Ala Ser Gln Gly Ile Ser Ser Ala Leu Val

1 5 10

<210> 209

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 326

<400> 209

Asp Ala Ser Ser Leu Glu Ser

1 5

<210> 210

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 326

<400> 210

Gln Gln Phe Asn Ser Tyr Pro Leu Thr

1 5

<210> 211

<211> 357

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 329

<400> 211

caggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cgcctttcgc aactatgcca tgaactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgttt attactgtgc gagagactac 300

ggtggtatct ccccctttga ctactggggc cagggaaccc tggtcactgt ctcctca 357

<210> 212

<211> 119

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 329

<400> 212

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Arg Asn Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Gly Gly Ile Ser Pro Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 213

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 329

<400> 213

Asn Tyr Ala Met Asn

1 5

<210> 214

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 329

<400> 214

Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 215

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 329

<400> 215

Asp Tyr Gly Gly Ile Ser Pro Phe Asp Tyr

1 5 10

<210> 216

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 329

<400> 216

gacatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt accctcacac ttttggccag 300

gggaccaaag tggatatcaa a 321

<210> 217

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 329

<400> 217

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro His

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Asp Ile Lys

100 105

<210> 218

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 329

<400> 218

Arg Ala Ser Gln Gly Ile Ser Ser Ala Leu Ala

1 5 10

<210> 219

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 329

<400> 219

Asp Ala Ser Ser Leu Glu Ser

1 5

<210> 220

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 329

<400> 220

Gln Gln Phe Asn Ser Tyr Pro His Thr

1 5

<210> 221

<211> 372

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG446

<400> 221

caggtgcagc tggtggagtc tgggggaggt gtggtacggc cgggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttgat gattatggca tgagctgggt ccgccaagct 120

ccagggaagg ggctggagtg ggtctctaat attaattgga atggtgatag cacaggttat 180

gtagactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240

ctgcaaatga acagtctgag agccgaggac acggccttgt attactgtgc gagagcgagg 300

acctattact atgtttcggg gagtactac tttgactact ggggccaggg aaccctggtc 360

actgtctcct ca 372

<210> 222

<211> 124

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG446

<400> 222

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Arg Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Asn Ile Asn Trp Asn Gly Asp Ser Thr Gly Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Arg Ala Arg Thr Tyr Tyr Tyr Val Ser Gly Arg Tyr Tyr Phe Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 223

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG446

<400> 223

Asp Tyr Gly Met Ser

1 5

<210> 224

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG446

<400> 224

Asn Ile Asn Trp Asn Gly Asp Ser Thr Gly Tyr Val Asp Ser Val Lys

1 5 10 15

Gly

<210> 225

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG446

<400> 225

Ala Arg Thr Tyr Tyr Tyr Val Ser Gly Arg Tyr Tyr Phe Asp Tyr

1 5 10 15

<210> 226

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG446

<400> 226

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt accgctcac tttcggcgga 300

gggaccaagg tggaaatcaa a 321

<210> 227

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG446

<400> 227

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 228

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR1 of MOG446

<400> 228

Arg Ala Ser Gln Gly Ile Ser Ser Ala Leu Ala

1 5 10

<210> 229

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR2 of MOG446

<400> 229

Asp Ala Ser Ser Leu Glu Ser

1 5

<210> 230

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR3 of MOG446

<400> 230

Gln Gln Phe Asn Ser Tyr Pro Leu Thr

1 5

<210> 231

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG456

<400> 231

gaggtgcagc tggtggagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatagca tgcactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcatcc attggtagta ggagtcgtta catatactac 180

gcagactcag tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gaaagggtat 300

tacgatattt tgactggttc tctctttgac tactggggcc agggaaccct ggtcactgtc 360

tcctca 366

<210> 232

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 456

<400> 232

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ser Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Gly Ser Arg Ser Arg Tyr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Tyr Tyr Asp Ile Leu Thr Gly Ser Leu Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 233

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG456

<400> 233

Ser Tyr Ser Met His

1 5

<210> 234

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG456

<400> 234

Ser Ile Gly Ser Arg Ser Arg Tyr Ile Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 235

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 456

<400> 235

Gly Tyr Tyr Asp Ile Leu Thr Gly Ser Leu Phe Asp Tyr

1 5 10

<210> 236

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG456

<400> 236

gacatcgtga tgacccagtc tccatcctca ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgtc gggcgagtca gggtattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt acccgtggac gttcggccaa 300

gggaccaagg tggagatcaa a 321

<210> 237

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG456

<400> 237

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 238

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 1 of MOG 456

<400> 238

Arg Ala Ser Gln Gly Ile Ser Ser Ala Leu Ala

1 5 10

<210> 239

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 456

<400> 239

Asp Ala Ser Ser Leu Glu Ser

1 5

<210> 240

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 456

<400> 240

Gln Gln Phe Asn Ser Tyr Pro Trp Thr

1 5

<210> 241

<211> 357

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 473

<400> 241

caggtgcagc tggtggagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatagca tgcactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtttcggac attagtacta atagtagaac cagaaactat 180

gcagactctg tgaagggccg attcaccatc tccagagaca atgccaagaa ctcagtgtat 240

ctgcaaatgc acagcctgag ggacgaggac acggctgtgt actactgtgc gagagactac 300

ggtggtatct attactttga ctattggggc cagggaaccc tggtcactgt ctcctca 357

<210> 242

<211> 119

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 473

<400> 242

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ser Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Asp Ile Ser Thr Asn Ser Arg Thr Arg Asn Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr

65 70 75 80

Leu Gln Met His Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Gly Gly Ile Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 243

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR1 of MOG 473

<400> 243

Ser Tyr Ser Met His

1 5

<210> 244

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR2 of MOG 473

<400> 244

Asp Ile Ser Thr Asn Ser Arg Thr Arg Asn Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 245

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of HCDR3 of MOG 473

<400> 245

Asp Tyr Gly Gly Ile Tyr Tyr Phe Asp Tyr

1 5 10

<210> 246

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 473

<400> 246

gaaatagtga tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccgta cacttttggc 300

caggggacca agctggagat caaa 324

<210> 247

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 473

<400> 247

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 248

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR1 of MOG 473

<400> 248

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 249

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 2 of MOG 473

<400> 249

Asp Ala Ser Asn Arg Ala Thr

1 5

<210> 250

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of LCDR 3 of MOG 473

<400> 250

Gln Gln Arg Ser Asn Trp Pro Pro Tyr Thr

1 5 10

<210> 251

<211> 369

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG426

<400> 251

caggtccagc tggtacagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggtta cagctttaac agctatggta tcaactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt aacactggtaa aacaagttat 180

gcacagaagg tccagggcag agtcaccatg accacagaca gatccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccatgt attactgtgc gagagagtac 300

gatattttga ctggttatattc cgatgctttt gatacctggg gccaagggac aatggtcacc 360

gtctcttca 369

<210> 252

<211> 123

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG426

<400> 252

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Asn Ser Tyr

20 25 30

Gly Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Thr Gly Lys Thr Ser Tyr Ala Gln Lys Val

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Arg Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Glu Tyr Asp Ile Leu Thr Gly Tyr Ser Asp Ala Phe Asp Thr

100 105 110

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 253

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG426

<400> 253

gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtggcaact ggccgctcac tttcggcgga 300

gggaccaagc tggagatcaa a 321

<210> 254

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG426

<400> 254

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Gly Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 255

<211> 369

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG428

<400> 255

gaggtccagc tggtacagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggtta cagctttaac agctatggta tcaactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt aacactggtaa aacaagttat 180

gcacagaagg tccagggcag agtcaccatg accacagaca gatccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccatgt attactgtgc gagagagtac 300

gatattttga ctggttatattc cgatgctttt gatacctggg gccaagggac aatggtcacc 360

gtctcttca 369

<210> 256

<211> 123

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG428

<400> 256

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Asn Ser Tyr

20 25 30

Gly Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Thr Gly Lys Thr Ser Tyr Ala Gln Lys Val

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Arg Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Glu Tyr Asp Ile Leu Thr Gly Tyr Ser Asp Ala Phe Asp Thr

100 105 110

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 257

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG428

<400> 257

gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttatgtcag cagtatggta gctcacctcg gacgttcggc 300

caagggacca agctggagat caaa 324

<210> 258

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG428

<400> 258

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 259

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 313

<400> 259

caggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagtt attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagcatat 300

tacgatattt tgactggttc cctctttgac tcctggggcc agggaaccct ggtcactgtc 360

tcctca 366

<210> 260

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 313

<400> 260

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Ala Tyr Tyr Asp Ile Leu Thr Gly Ser Leu Phe Asp Ser Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 261

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 313

<400> 261

gacatcgtga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt accctctcac tttcggcgga 300

gggaccaagg tggaaatcaa a 321

<210> 262

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG

<400> 262

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 263

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 314

<400> 263

caggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gaggtagtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaaggctat 300

tacgatattt tgactggttc cttctttgac tactggggcc agggaaccct ggtcaccgtc 360

tcctca 366

<210> 264

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 314

<400> 264

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Tyr Tyr Asp Ile Leu Thr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 265

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 314

<400> 265

gacatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt acccgtacac ttttggccag 300

gggaccaagc tggagatcaa a 321

<210> 266

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 314

<400> 266

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 267

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 315

<400> 267

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc acctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtgttag cacatactac 180

gcagactccg tgaagggccg gttcaccctc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagcgtat 300

tacgatattt tgactggtaa tttccttgac tactggggcc agggaaccct ggtcactgtc 360

tcctca 366

<210> 268

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 315

<400> 268

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Val Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Leu Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Ala Tyr Tyr Asp Ile Leu Thr Gly Asn Phe Leu Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 269

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 315

<400> 269

gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt accgctcac tttcggcgga 300

gggaccaagg tggaaatcaa a 321

<210> 270

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 315

<400> 270

Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 271

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG331

<400> 271

gaggtgcagc tggtggagtc cgggggaggc ttggtatagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagcgtat 300

tacgatattt tgactggttc cttctttgac tactggggcc agggaaccct ggtcactgtc 360

tcctca 366

<210> 272

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG331

<400> 272

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Tyr Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Ala Tyr Tyr Asp Ile Leu Thr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 273

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG331

<400> 273

gaaatagtga tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag actggagcct 240

gaagattttg cagtgtatta ctgtcagcag tatggtagct caccgctcac tttcggcgga 300

gggaccaagc tggagatcaa a 321

<210> 274

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG331

<400> 274

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 275

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG357

<400> 275

gaggtgcagc tggtggagac tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagat attaatcata gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagca cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagactat 300

tacgatattt tgactggttc cttctttgac tactggggcc agggaaccct ggtcactgtc 360

tcctca 366

<210> 276

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 357

<400> 276

Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Asp Ile Asn His Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys His Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Asp Tyr Tyr Asp Ile Leu Thr Gly Ser Phe Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 277

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 357

<400> 277

gacatcgtga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt acccgtggac gttcggccaa 300

gggaccaagg tggagatcaa a 321

<210> 278

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 357

<400> 278

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 279

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 476

<400> 279

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacttttagc agctatgcca tgaactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagttata gtggtcgtag cacatactac 180

gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaagggcctt 300

tacgatattt tgactggtgg cggatttgac tactggggcc agggaaccct ggtcaccgtc 360

tcttca 366

<210> 280

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 476

<400> 280

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Tyr Ser Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Leu Tyr Asp Ile Leu Thr Gly Gly Gly Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 281

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 476

<400> 281

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcaacag tttaatagtt acccgtccac ttttggccag 300

gggacacgac tggagattaa a 321

<210> 282

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 476

<400> 282

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Ser

85 90 95

Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 283

<211> 381

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 323

<400> 283

cggatcacct tgagggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60

acctgcacct tctctgggtt ctcactcagc actagtggag tgggtgtggg ctggatccgt 120

cagcccccag gaaaggccct ggagtggctt gcactcattt tttggggatga tgatagtcac 180

tacagcccat ctctgaagag caggctcacc atcaccaagg acacctccaa aaaccaggtg 240

gtccttacaa tgaccaacat ggaccctgtg gacacagcca catattactg tgcacggtat 300

tactttggtt cggggagtta tttccctagc tactggtact tcgatctctg gggccgtggc 360

accctggtca ccgtctcctc a 381

<210> 284

<211> 127

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 323

<400> 284

Arg Ile Thr Leu Arg Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser

20 25 30

Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu

35 40 45

Trp Leu Ala Leu Ile Phe Trp Asp Asp Asp Ser His Tyr Ser Pro Ser

50 55 60

Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val

65 70 75 80

Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Ala Arg Tyr Tyr Phe Gly Ser Gly Ser Tyr Phe Pro Ser Tyr Trp

100 105 110

Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 285

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 323

<400> 285

gaaatagtga tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctcccac tttcggcgga 300

gggaccaagc tggagatcaa a 321

<210> 286

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 323

<400> 286

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 287

<211> 381

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 341

<400> 287

cggatcacct tgaaggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60

acctgcacct tctctgggtt ctcactcagc actagtggag tgggtgtggg ctggatccgt 120

cagcccccag gaaaggccct ggagtggctt gcactcattt attgggatga tgataaacac 180

tacagcccat ctctgaagag caggctcacc atcaccaagg acacctccaa aaaccaggtg 240

gtccttacaa tgaccaacat ggaccctgtg gacacagcca catattactg tgcacggtat 300

tactttggtt cggggagtta ttcccctagc tactggtact tcgatctctg gggccgtggc 360

accctggtca ctgtctcctc a 381

<210> 288

<211> 127

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 341

<400> 288

Arg Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser

20 25 30

Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu

35 40 45

Trp Leu Ala Leu Ile Tyr Trp Asp Asp Asp Lys His Tyr Ser Pro Ser

50 55 60

Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val

65 70 75 80

Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Ala Arg Tyr Tyr Phe Gly Ser Gly Ser Tyr Ser Pro Ser Tyr Trp

100 105 110

Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 289

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 341

<400> 289

gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctcccac tttcggcgga 300

gggaccaagg tggagatcaa a 321

<210> 290

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 341

<400> 290

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 291

<211> 381

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 354

<400> 291

cggatcacct tgagggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60

acctgcacct cctctgggct ctcactcagc actagtggag tgggtgtggg ctggatccgt 120

cagcccccag gaaaggccct ggagtggctt gcactcattt tttggggatga tgatacacac 180

tacagcccat ctctgaagag caggctcacc atcaccaagg acacctccaa aaaccaggtg 240

gtccttacaa tgaccaacat ggaccctgtg gacacagcca catattactg tgcacggtat 300

tactttggtt cggggagtta tttccctagc tactggtact tcgatctctg gggccgtggc 360

accatggtca ccgtctcttc a 381

<210> 292

<211> 127

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 354

<400> 292

Arg Ile Thr Leu Arg Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Ser Ser Gly Leu Ser Leu Ser Thr Ser

20 25 30

Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu

35 40 45

Trp Leu Ala Leu Ile Phe Trp Asp Asp Asp Thr His Tyr Ser Pro Ser

50 55 60

Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val

65 70 75 80

Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Ala Arg Tyr Tyr Phe Gly Ser Gly Ser Tyr Phe Pro Ser Tyr Trp

100 105 110

Tyr Phe Asp Leu Trp Gly Arg Gly Thr Met Val Thr Val Ser Ser

115 120 125

<210> 293

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 354

<400> 293

gaaatagtga tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgctcac tttcggcgga 300

gggaccaagg tggaaatcaa a 321

<210> 294

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 354

<400> 294

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 295

<211> 381

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 355

<400> 295

cggatcacct tgagggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60

acctgcacct cctctgggct ctcactcagc actagtggag tgggtgtggg ctggatccgt 120

cagcccccag gaaaggccct ggagtggctt gcactcattt tttggggatga tgatacacac 180

tacagcccat ctctgaagag caggctcacc atcaccaagg acacctccaa aaaccaggtg 240

gtccttacaa tgaccaacat ggaccctgtg gacacagcca catattactg tgcacggtat 300

tactttggtt cggggagtta tttccctagc tactggtact tcgatctctg gggccgtggc 360

accctggtca ctgtctcctc a 381

<210> 296

<211> 127

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 355

<400> 296

Arg Ile Thr Leu Arg Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Ser Ser Gly Leu Ser Leu Ser Thr Ser

20 25 30

Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu

35 40 45

Trp Leu Ala Leu Ile Phe Trp Asp Asp Asp Thr His Tyr Ser Pro Ser

50 55 60

Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val

65 70 75 80

Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr

85 90 95

Cys Ala Arg Tyr Tyr Phe Gly Ser Gly Ser Tyr Phe Pro Ser Tyr Trp

100 105 110

Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 297

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 355

<400> 297

gaaatagtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgctcac tttcggcgga 300

gggaccaagc tggagatcaa a 321

<210> 298

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 355

<400> 298

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 299

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 308

<400> 299

gaggtgcagc tggtgcagtc cggggctgag gtgaggaagt ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttagtac gacaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggaactga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatggggtt tgcctactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 300

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 308

<400> 300

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Arg Lys Ser Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Ser Thr Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Ala Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 301

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 308

<400> 301

gaaatagtga tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccgac gttcggccaa 300

gggaccaaag tggatatcaa a 321

<210> 302

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 308

<400> 302

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Asp Ile Lys

100 105

<210> 303

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 316

<400> 303

gaagtgcagc tggtgcagtc cggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttaatac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatggggtt taactactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 304

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 316

<400> 304

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Asn Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asn Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 305

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 316

<400> 305

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattaac agtgctttag cctggtatca gcagaaacca 120

gggaaagctc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgccaacag tataatagtt acccgatcac cttcggccaa 300

gggacacgac tggagattaa a 321

<210> 306

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 316

<400> 306

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asn Ser Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Ile

85 90 95

Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 307

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 319

<400> 307

caggtgcagc tggtgcaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttagtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgacgac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatgggctt taactactgg ggccagggaa ccctggtcac tgtctcctca 360

<210> 308

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 319

<400> 308

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Ser Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asn Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 309

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 319

<400> 309

gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccatt cactttcggc 300

cctgggacca aggtggaaat caaa 324

<210> 310

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 319

<400> 310

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Phe Thr Phe Gly Pro Gly Thr Lys Val Glu Ile Lys

100 105

<210> 311

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 320

<400> 311

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcaactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttagtac agtaaattac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatggggtt tgactactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 312

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 320

<400> 312

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Ser Thr Val Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 313

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 320

<400> 313

gaaatagtgt tgacacagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccatt cactttcggc 300

cctgggacca aagtggatat caaa 324

<210> 314

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 320

<400> 314

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Phe Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 315

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 338

<400> 315

caggtccagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttaatac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatggggtt tgacccctgg ggccagggaa ccctggtcac tgtctcctca 360

<210> 316

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 338

<400> 316

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Asn Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asp Pro Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 317

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 338

<400> 317

gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag cgtaacaact ggcctctcac tttcggcgga 300

gggaccaagc tggagatcaa a 321

<210> 318

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 338

<400> 318

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Asn Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 319

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG352

<400> 319

gaggtgcagc tggtgcagtc cggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcaactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttagtac agtaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatggggtt tgactactgg ggccagggaa ccctggtcac tgtctcctca 360

<210> 320

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG352

<400> 320

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Ser Thr Val Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 321

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG352

<400> 321

gccatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc 60

atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcataaacca 120

gggaaagccc ctaagtccct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgccaacag tataatagtt accgctcac tttcggcgga 300

gggaccaaag tggatatcaa a 321

<210> 322

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 352

<400> 322

Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys

100 105

<210> 323

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 359

<400> 323

gaggtccagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttaatac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagattgg 300

gcagtggctg gtatggggtt taactactgg ggccagggaa ccctggtcac tgtctcctca 360

<210> 324

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 359

<400> 324

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Asn Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Gly Met Gly Phe Asn Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 325

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 359

<400> 325

gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccgct cactttcggc 300

ggagggacca agctggagat caaa 324

<210> 326

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 359

<400> 326

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 327

<211> 360

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 478

<400> 327

gaggtccagc tggtacagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agatatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtttgctac agcaaactac 180

gcacagaagt tccaggccag agtcacgatt accgcggacg aaaccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acgggcgtgt attactgtgc gagagattgg 300

gcagtggctg ctatggggtt tgcccactgg ggccagggaa ccctggtcac tgtctcctca 360

<210> 328

<211> 120

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 478

<400> 328

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Arg Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Met Phe Ala Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Ala Arg Val Thr Ile Thr Ala Asp Glu Thr Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Gly Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ala Val Ala Ala Met Gly Phe Ala His Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 329

<211> 324

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 478

<400> 329

gaaatagtga tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120

cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180

gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 240

cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccgat caccttcggc 300

caagggacac gactggagat taaa 324

<210> 330

<211> 108

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 478

<400> 330

Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 331

<211> 357

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 470

<400> 331

caggtgcagc tggtggagtc tgggggaggc ttggtacagc cgggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgaactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaagtga acagcctgag agccgaggac acggccgttt attactgtgc gagagactac 300

ggtggtatct ccccctttga ctactggggc cagggaaccc tggtcaccgt ctcctca 357

<210> 332

<211> 119

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 470

<400> 332

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Val Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Gly Gly Ile Ser Pro Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 333

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 470

<400> 333

gaaatagtga tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag actggagcct 240

gaagattttg cagtgtatta ctgtcagcag tatggtagct caccgtacac ttttggccag 300

gggacacgac tggagattaa a 321

<210> 334

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 470

<400> 334

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 335

<211> 366

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VH excluding signal sequence of MOG 418

<400> 335

caggtgtagc tggtgcagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatagca tgcactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcatcc attagtcata gtagtagtta catatcctac 180

gcagactcag tgaagggccg attcaccatc tccagagaca acgccaagaa ttcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gaaagggtat 300

tacgatattt tgactggttc tctctttgac tactggggcc agggaaccct ggtcaccgtc 360

tcctca 366

<210> 336

<211> 122

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VH excluding signal sequence of MOG 418

<400> 336

Gln Val Tyr Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ser Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser His His Ser Ser Ser Tyr Ile Ser Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Tyr Tyr Asp Ile Leu Thr Gly Ser Leu Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 337

<211> 321

<212> DNA

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: base sequence

encoding VL excluding signal sequence of MOG 418

<400> 337

gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag actggagcct 240

gaagattttg cagtgtatta ctgtcagcag tatggtagct caccattcac tttcggccct 300

gggaccaaag tggatatcaa a 321

<210> 338

<211> 107

<212> PRT

<213> Artificial Sequence

<220>

<223> The Description of the artificial sequence: amino acid sequence

of VL excluding signal sequence of MOG 418

<400> 338

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

Claims (18)

1. An antibody or an antigen-binding fragment thereof, which binds to the myelin oligodendrocyte glycoprotein MOG, wherein the antibody or the antigen-binding fragment thereof comprises: The complementarity-determining regions, namely the heavy chain variable regions (VH) whose amino acid sequences of CDR1–3 are composed of the amino acid sequences recorded in sequence numbers 4, 5, and 6, respectively, and the light chain variable regions (VL) whose amino acid sequences of CDR1–3 are composed of the amino acid sequences recorded in sequence numbers 10, 11, and 12, respectively. 2. The antibody or its antigen-binding fragment as described in claim 1, wherein the antibody has brain retention properties. 3. The antibody or its antigen-binding fragment as described in claim 1 or 2, wherein the antibody is: The VH antibody consists of the amino acid sequence described in sequence number 3, and the VL antibody consists of the amino acid sequence described in sequence number 9. 4. The antibody or antigen-binding fragment thereof as described in claim 1 or 2, wherein the antibody is a bispecific antibody. 5. The antibody or antigen-binding fragment thereof as claimed in claim 4, wherein the bispecific antibody binds to the MOG and the antigen present in the brain. 6. The antibody or antigen-binding fragment thereof as claimed in claim 4, wherein the bispecific antibody comprises an antigen-binding site that binds to MOG and an antigen-binding site that binds to antigens present in the brain. 7. The antibody or antigen-binding fragment thereof as described in claim 1 or 2, wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab') ₂ , single-chain antibody (scFv), dimerized V region (double-chain antibody), and disulfide-stabilized V region (dsFv). 8. The antibody or antigen-binding fragment thereof as described in claim 1 or 2, wherein the antibody is a recombinant antibody. 9. The antibody or antigen-binding fragment thereof as described in claim 1 or 2, wherein the antibody is a human antibody. 10. A fusion antibody or fusion antibody fragment, formed by conjugating at least one selected from the group consisting of (a) to (c) below with an antibody or antigen-binding fragment thereof that binds to MOG as described in any one of claims 1 to 9. (a) Hydrophilic polymers, (b) Amphiphilic macromolecules, and (c) Functional molecules. 11. A cell that produces the antibody or antigen-binding fragment thereof as described in any one of claims 1 to 9, or the fusion antibody or fusion antibody fragment as described in claim 10. 12. A nucleic acid comprising a base sequence encoding an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 9, or a fusion antibody or fusion antibody fragment as described in claim 10. 13. A transformed cell containing a vector comprising the nucleic acid of claim 12. 14. A method for manufacturing the antibody or antigen-binding fragment thereof of any one of claims 1 to 9, or the fusion antibody or fusion antibody fragment thereof of claim 10, comprising: culturing the cells of claim 11 or the transformed cells of claim 13, and collecting the antibody or antigen-binding fragment thereof of any one of claims 1 to 9, or the fusion antibody or fusion antibody fragment thereof of claim 10, from the culture medium. 15. A composition comprising the antibody or antigen-binding fragment thereof as described in any one of claims 1 to 9, or the fusion antibody or fusion antibody fragment as described in claim 10. 16. The composition of claim 15, wherein it is a composition for detecting or determining antigens present in the brain. 17. The composition of claim 15, wherein it is a composition for diagnosing or treating brain diseases. 18. Use of the antibody or antigen-binding fragment thereof of any one of claims 1 to 9, the fusion antibody or fusion antibody fragment of claim 10, or the composition of claim 15 in the preparation of a diagnostic composition for brain diseases expressing MOG in the brain or a pharmaceutical composition for treating brain diseases expressing MOG in the brain.