Antibody that binds to cell adhesion molecule 3

Abstract

The present invention relates to: an antibody that binds to cell adhesion molecule 3 (CADM3) or a fragment of the antibody, a hybridoma that produces the antibody or antibody fragment, a nucleic acid containing a base sequence encoding the antibody or antibody fragment, a transformed cell containing a vector containing the nucleic acid, a method for producing the antibody or antibody fragment, a composition containing the antibody or antibody fragment, a method for detecting or measuring an antigen present in the brain by using the antibody or antibody fragment, a method for diagnosing or treating a brain disease, a method for improving the retention of an antibody in the brain, a method for increasing the amount of an antibody in the brain, and the like.

Description

Technical Field

[0001] The present invention relates to, for example, antibodies or antibody fragments thereof that bind to cell adhesion molecule 3 (CADM3), hybridomas that produce the antibodies or antibody fragments thereof, nucleic acids comprising a nucleotide sequence encoding the antibodies or antibody fragments thereof, transformant cells comprising a vector comprising the nucleic acid, methods for producing the antibodies or antibody fragments thereof, compositions comprising the antibodies or antibody fragments thereof, methods for detecting or measuring antigens present in the brain, methods for diagnosing or treating brain diseases, methods for improving the properties of antibodies that accumulate in the brain, and methods for increasing the amount of antibodies in the brain, each of which uses the antibodies or antibody fragments thereof. Background Art

[0002] Many antibody drugs have been developed since the FDA approved the mouse anti-CD3 antibody muromonab-CD3 (OKT3) as the first antibody drug in 1986. In 1994, the chimeric antibody abciximab was approved, in which the variable region of a mouse antibody and the constant region of a human antibody were linked to reduce the antigenicity of the mouse antibody.

[0003] In order to further reduce antigenicity, humanized antibody technology was developed, in which the complementarity determining regions (CDRs) of the mouse antibody variable region, which play an important role in binding to the antigen, were transplanted into the framework regions (FRs) of human antibodies, and the humanized anti-CD20 antibody dacizumab was approved in 1997.

[0004] In addition, phage display technology using human antibody sequence libraries has been used, and the fully human anti-TNF-α antibody adalimumab was approved as the first antibody obtained using phage display technology in 2002. More than 60 antibody drugs targeting antigens such as CD20, CD52, TNF-α, HER2, and EGFR have been approved (NPL 1).

[0005] In this way, antibodies have become a widely accepted drug format. Most of the antibody drugs approved so far are for cancer and immune diseases, accounting for more than 75% of all antibody drugs.

[0006] Biologics, such as antibodies, are becoming increasingly important in the treatment of central nervous system diseases. A monoclonal antibody targeting amyloid beta has been reported for use in Alzheimer's disease, and various types of neurotrophic factors with neuroprotective effects (brain-derived neurotrophic factor (BDNF) and glial cell-derived neurotrophic factor (GDNF)) have demonstrated neuroprotective effects in central nervous system diseases in animal models (NPL 2).

[0007] However, when antibodies are administered peripherally, the amount delivered to the central nervous system is lower than that delivered to other organs, and the antibody mobility (ratio of concentration in cerebrospinal fluid (CSF) to serum concentration) is reported to be 0.1 to 0.3% (NPL 3 to 5).

[0008] The reason for the reduced drug delivery in the central nervous system, which includes the brain and bone marrow, is a mechanism known as the blood-brain barrier (BBB), which restricts the transport of substances between the blood and the interstitial fluid of the brain. The BBB has a physical / nonspecific control mechanism caused by intercellular adhesion of vascular endothelial cells and a substrate-specific efflux mechanism caused by efflux transporters, protecting the central nervous system from the effects of foreign substances or drugs and playing an important role in maintaining homeostasis.

[0009] However, due to the existence of blood-brain barrier, it is not easy to obtain the effective concentration in the central nervous system when administering, and drug development is difficult.For example, although by to Hurler syndrome (I type mucopolysaccharidosis) intravenous administration α-L-iduronidase or to Hunter syndrome (II type mucopolysaccharidosis) intravenous administration iduronic acid-2-sulfatase, enzyme replacement therapy has been carried out, but the enzyme does not pass through the blood-brain barrier due to molecular weight, therefore the effect for central nervous system symptoms is not observed (NPL 6 to 9). In addition, it is reported that due to a certain amount of recombinase being regularly administered continuously, the generation (NPL 10) of side effect such as neutralizing antibodies has been caused.

[0010] In addition, attempts to directly administer biological products into the bone marrow cavity or brain have also been carried out to increase the concentration in the brain. For example, a method of administering iduronate-2-sulfatase to the brain of a patient with Hunter syndrome (mucopolysaccharidosis type II) to prevent the progression of the patient's brain disorder has been reported (PTL 1). However, direct administration into the bone marrow cavity or brain is highly invasive (NPL 11).

[0011] Therefore, various delivery technologies have been studied to increase the concentration of high molecular weight substances such as biological products in the brain. For example, a method has been reported in which a high molecular weight substance is bound to a membrane protein expressed in cerebral vascular endothelial cells to form a complex of the substance and the membrane protein, and the complex is allowed to pass through the blood-brain barrier by endocytosis.

[0012] Most of the reported techniques use receptor-mediated transcytosis (RMT), and receptors expressed in brain vascular endothelial cells serving as targets include, for example, transferrin receptor, insulin receptor, insulin-like growth factor receptor, low-density lipoprotein receptor family (LDLRf), and the like.

[0013] A technology for crossing the blood-brain barrier via the transferrin receptor by producing a fusion protein of an anti-transferrin receptor antibody and nerve growth factor has been reported. Technologies using anti-transferrin receptor antibodies include bispecific antibodies combining an anti-transferrin receptor antibody and an anti-β-secretase (BACE1) antibody (PTLs 2 and 3 and NPLs 12 and 13), and fusion antibodies obtained by fusing a monovalent anti-transferrin receptor antibody to the carboxyl terminus of an anti-amyloid peptide β antibody (PTL 4 and NPL 14).

[0014] It has been reported that for brain delivery of a bispecific antibody using an anti-transferrin receptor antibody and an anti-BACE1 antibody, when the antibody was administered to mice at a dose of 20 mg / kg body weight, the amount of the antibody incorporated into the brain increased approximately 4-fold compared to the control (NPL 13).

[0015] In addition, a technique for allowing a drug to pass through the blood-brain barrier by encapsulating it in liposomes having anti-transferrin receptor antibodies on their surfaces has been reported. It has been reported that the amount incorporated into the rat brain was increased by about 2 to 5 times by a fusion of anti-rat transferrin receptor antibodies and immune micelles (NPL 15).

[0016] Furthermore, a technique for passing the blood-brain barrier via the insulin receptor by producing a fusion protein in which a neurotrophic factor, an enzyme, or an anti-amyloid peptide antibody is fused to the carboxyl terminus of an anti-insulin receptor antibody has been reported (NPL 16 to 19).

[0017] It has been reported that in rhesus monkeys, 2 hours after administration of a labeled anti-human insulin receptor antibody and GDNF fusion antibody, the amount incorporated into the brain was approximately 15 times that of GDNF (NPL 17).

[0018] However, transferrin receptors and insulin receptors are expressed not only in cerebral vascular endothelial cells but also throughout the body, including the liver, and therefore, when the amount of drug delivered to the central nervous system increases with these techniques, the drug is also delivered to the liver, etc. (NPL 20). In addition, since the antigen is expressed throughout the body, the half-life of the antibody in the blood is short (NPL 12).

[0019] In addition, it has been reported that an antibody (Fc5) against TMEM30A, an antigen expressed on the membrane of brain vascular endothelial cells, exhibits RMT-like activity (PTL 5 and NPLs 21 and 22). Fc5 is an antibody derived from the heavy chain variable domain (VHH) of a single-domain heavy chain antibody from a llama, and in vitro BBB models and in vivo rat models have demonstrated that the amount of Fc5 and human Fc fusion delivered to the brain is increased compared to control IgG.

[0020] It was reported that in a rat model, a fusion of an Fc5-derived single-chain antibody (scFv) and a type I metabotropic glutamate receptor (mGluRI) antibody increased CSF exposure compared to a fusion of a control single-chain antibody and an mGluRI antibody, but the increase was approximately 5-fold (NPL 23).

[0021] It has also been reported that IgG antibodies are rapidly excreted from the brain into the circulation via the neonatal Fc receptor (FcRn) (NPLs 24 and 25), and that, for example, in rats, the half-life of IgG in the brain is as short as 48 minutes after administration into the brain (NPL 24).

[0022] CADM3 is a calcium-independent immunoglobulin-like cell adhesion molecule (NPLs 26 to 31). CADM3 is divided into multiple structures, including three immunoglobulin-like domains as extracellular domains, a transmembrane domain, and a cytoplasmic domain (NPL 29).

[0023] From the perspective of Northern blot and in situ hybridization analysis, CADM3 is specifically expressed in various central and peripheral nerve tissues including the cerebellum, cerebral cortex, hippocampus, amygdala, olfactory bulb and medulla oblongata (NPL 26, 27 and 32). CADM3 is located at the contact sites between two axon terminals, between axon terminals and axon shafts, and between axon terminals and glial cell processes at axon terminals (NPL 26).

[0024] CADM3 exhibits cell-cell adhesion activity by calcium ion-independent homophilic binding. In addition, CADM3 exhibits cell-cell adhesion activity by calcium ion-independent homophilic binding with Necl-2, connexin-1 and connexin-3, but does not exhibit adhesion activity with Necl-5 and connexin-2. CADM3 interacting with connexin-1 and connexin-3 participates in the neuronal activity-dependent synaptic remodeling process in the same manner as in the formation of cerebellar morphology (NPL 32 and 33). In vitro binding analysis confirmed that protein 4.1N involved in actin cytoskeleton rearrangement binds to CADM3 (NPL 27).

[0025] In CADM3 knockout mice, the number of myelinated axons in the optic nerve and spinal cord is reduced in the early postnatal period. However, there is no difference in the number of myelinated axons or the thickness of the myelin sheath between normal individuals and mutants after maturity (NPL 30). In addition, a polyclonal antibody that binds to CADM3 has been reported (NPL 27).

[0026] Citation List

[0027] Patent Literature

[0028] PTL 1: WO 2012 / 023623

[0029] PTL 2: WO 2016 / 081640

[0030] PTL 3: WO 2016 / 081643

[0031] PTL 4: WO 2014 / 033074

[0032] PTL 5: Canadian Patent No. 2623841

[0033] Non-patent literature

[0034] NPL 1: Kyla RR. and Richard CC., Biotechnol Adv, pii: S0734-9750(16), 30091-X, 2016

[0035] NPL 2: Pardridge WM., Bioconjugate Chem., 19, 1327-1338, 2008

[0036] NPL 3: Wang W. et al., Clin.pharmacol.Ther., 84, 548-558, 2008

[0037] NPL 4: Garg A. et al., AAPSJ., 11, 553-557, 2009

[0038] NPL 5: Kaj B. et al., Arch. Neurol., 69(8), 1002-1010, 2012

[0039] NPL 6: Wraith JE. et al., J.Pediatr.144(5), 581-588, 2004

[0040] NPL 7: Muenzer J. et al., Genet Med. 8(8), 465-473, 2006

[0041] NPL 8: Package insert for 2.9 mg Aldurazyme (registered trademark) intravenous infusion solution (July 2016, 8th edition)

[0042] NPL 9: Package insert for 6 mg Elaprase (registered trademark) intravenous infusion (July 2016, 6th edition)

[0043] NPL 10: Brooks, DA et al., Trends Mol. Med. 9, 450-453, 2003

[0044] NPL 11: Sorrentino NC., et al., Pediatr Endocrinol Rev. 1, 630 - 638, 2016

[0045] NPL12: Couch JA., et al., Science Translational Medicine, 5, 183ra57, 2013

[0046] NPL 13: Yu YJ., et al., Science Translational Medicine, 6, 261ra154, 2014

[0047] NPL 14: Niewoehner J., et al., Neuron. 81, 49 - 60, 2014

[0048] NPL 15: Jun Y., et al., Macromol. Biosci. 12, 1209 - 1219, 2012

[0049] NPL 16: Pardridge WM. and Boado RJ., Methods in Enzymology, 503, 269 - 292, 2012

[0050] NPL 17: Boado RJ., et al., Drug Metab. Dispos., 37(12), 2299 - 2304, 2009

[0051] NPL 18: Boado RJ., et al., J. Pharmacol. Exp. Ther., 333(3), 961 - 969, 2010

[0052] NPL 19: Boado RJ., et al., Bioconjugate Chem., 1, 97 - 104, 2012

[0053] NPL 20: Yun Zhang., et al., J. Pharmacol. Exp. Ther., 313(3), 1075 - 1081.2005

[0054] NPL 21: Abulrob A., et al., J. Neuyrochem., 95(4), 1201 - 1214, 2005

[0055] NPL 22: Farrington GK., et al., FASEB J., 28, 4764 - 4778, 2014

[0056] NPL 23: Webster CI. et al., FASEB J., 30, 1927-1940, 2016

[0057] NPL 24: Zhang Y. et al., J. Neuroimmunol., 114(1-2), 168-172, 2001

[0058] NPL 25: Philip RC. et al., Brain Research, 1534, 13-21, 2013

[0059] NPL 26: Kakunaga S. et al., J. Cell Science, 118, 1267-1277, 2005

[0060] NPL 27: Zhou Y. et al., Biochim. Biophys. Acta, 1669, 142-154, 2005

[0061] NPL 28: Gao J. et al., Biochim. Biophys. Acta, 1778, 1429-1435, 2008

[0062] NPL 29: Dong X. et al., J. Biol. Chem., 281, 10610-10617, 2006

[0063] NPL 30: Park J. et al., J. Neurosci., 28, 12815-12819, 2008

[0064] NPL 31: Gruber-Olipitz M. et al., Amino Acids, 30, 409-415, 2006

[0065] NPL 32: Takai Y. et al., Cancer Sci., 94, 655-667, 2003

[0066] NPL 33: Sakisaka T. et al., Curr. Opin. Cell. Biol., 16, 513-521, 2004 Summary of the Invention

[0067] Technical issues

[0068] The present invention relates to, for example, CADM3-binding molecules that bind to CADM3, methods for using the same, and the like. Specifically, the present invention provides antibodies or antibody fragments thereof that bind to CADM3, hybridomas that produce the same, nucleic acids comprising nucleotide sequences encoding the same, transformant cells comprising vectors containing the same, methods for producing the same, compositions comprising the same, methods for detecting or measuring antigens present in the brain, methods for diagnosing or treating brain diseases, methods for improving the ability of antibodies to accumulate in the brain, and methods for increasing the amount of antibodies in the brain, each of which uses the same.

[0069] Solution to the problem

[0070] As a means for solving the above problems, the present invention provides a CADM3-binding molecule that binds to CADM3 and a method of using the molecule, specifically, an antibody or an antibody fragment thereof that binds to CADM3.

[0071] That is, the present invention relates to the following <1> to <22> .

[0072] <1> An antibody or antibody fragment thereof that binds to cell adhesion molecule 3 (CADM3).

[0073] <2> according to <1> The antibody or antibody fragment thereof, wherein the antibody has the property of accumulating in the brain.

[0074] <3> according to <1> or <2> The antibody or antibody fragment thereof, wherein the antibody has affinity for neurons and / or neural tissue.

[0075] <4> according to <1> to <3> The antibody or antibody fragment thereof according to any one of the preceding claims, wherein the antibody or antibody fragment thereof is selected from the following (a) to (x):

[0076] (a) an antibody, wherein the amino acid sequences of complementarity determining regions (CDRs) 1 to 3 of the heavy chain variable domain (VH) comprise the amino acid sequences represented by SEQ ID NOs: 23, 24, and 25, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of the light chain variable domain (VL) comprise the amino acid sequences represented by SEQ ID NOs: 28, 29, and 30, respectively;

[0077] (b) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 34, 35, and 36, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 38, 39, and 40, respectively;

[0078] (c) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of the heavy chain variable domain (VHH) of the heavy chain antibody comprise the amino acid sequences represented by SEQ ID NOs: 3, 4, and 5, respectively;

[0079] (d) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 8, 9, and 10, respectively;

[0080] (e) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 13, 14, and 15, respectively;

[0081] (f) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 18, 19, and 20, respectively;

[0082] (g) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 89, 90, and 91, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 94, 95, and 96, respectively;

[0083] (h) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 99, 100, and 101, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0084] (i) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 104, 105, and 106, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0085] (j) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 109, 110, and 111, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0086] (k) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 114, 115, and 116, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0087] (1) An antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 119, 120, and 121, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0088] (m) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 124, 125, and 126, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0089] (n) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 129, 130, and 131, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0090] (o) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 139, 140, and 141, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0091] (p) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 144, 145, and 146, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0092] (q) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 149, 150, and 151, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0093] (r) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 154, 155, and 156, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0094] (s) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 159, 160, and 161, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0095] (t) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 169, 170, and 171, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 174, 175, and 176, respectively;

[0096] (u) an antibody that competes with at least one of the antibodies or antibody fragments described in (a) to (t) for binding to CADM3;

[0097] (v) an antibody that binds to an epitope comprising the epitope to which any one of the antibodies or antibody fragments described in (a) to (t) binds;

[0098] (w) an antibody that binds to the same epitope as that bound by any one of the antibodies or antibody fragments described in (a) to (t); and

[0099] (x) An antibody comprising an amino acid sequence having 85% or higher homology to the amino acid sequence of any one of the antibodies or antibody fragments described in (a) to (t).

[0100] <5> according to <1> to <4> The antibody or antibody fragment thereof according to any one of the preceding claims, wherein the antibody or antibody fragment thereof is selected from the following (1) to (31):

[0101] (1) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 22, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 27;

[0102] (2) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 32, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 37;

[0103] (3) an antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 2;

[0104] (4) an antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 7;

[0105] (5) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 12;

[0106] (6) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 17;

[0107] (7) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 68;

[0108] (8) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 70;

[0109] (9) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 72;

[0110] (10) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 74;

[0111] (11) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 76;

[0112] (12) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 78;

[0113] (13) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 80;

[0114] (14) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 82;

[0115] (15) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 84;

[0116] (16) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 86;

[0117] (17) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 88, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 93;

[0118] (18) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 98, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0119] (19) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 103, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0120] (20) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 108, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0121] (21) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 113, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0122] (22) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 118, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0123] (23) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 123, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0124] (24) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 128, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0125] (25) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 138, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0126] (26) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 143, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0127] (27) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 148, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0128] (28) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 153, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0129] (29) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 158, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0130] (30) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 168, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 173; and

[0131] (31) An antibody comprising an amino acid sequence having 85% or higher homology to the amino acid sequence of any one of the antibodies or antibody fragments described in (1) to (30).

[0132] <6> according to <1> to <5> The antibody or antibody fragment thereof according to any one of claims 1 to 5, wherein the antibody or antibody fragment thereof is a bispecific antibody.

[0133] <7> according to <6> The bispecific antibody, wherein the bispecific antibody binds to CADM3 and an antigen present in the brain.

[0134] <8> according to <6> or <7> The bispecific antibody comprises an antigen binding site that binds to CADM3 and an antigen binding site that binds to an antigen present in the brain.

[0135] <9> according to <1> to <8> The antibody fragment of any one of the foregoing, wherein the antibody fragment is selected from Fab, Fab', F(ab')2, single-chain antibody (scFv), dimerized V region (diabody), disulfide-stabilized V region (dsFv), VHH and CDR-containing peptide.

[0136] <10> according to <1> to <9> The antibody and antibody fragment thereof according to any one of claims 1 to 5, wherein the antibody is a genetically recombinant antibody.

[0137] <11> according to <1> to <10> The antibody and antibody fragment thereof according to any one of the preceding claims, 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.

[0138] <12> A fusion antibody or a fusion antibody fragment thereof, wherein at least one selected from the following (i) to (iii) is linked to a <1> to <11> An antibody or antibody fragment thereof that binds to CADM3 according to any one of the following:

[0139] (i) a hydrophilic polymer;

[0140] (ii) an amphiphilic polymer; and

[0141] (iii) Functional molecules.

[0142] <13> A hybridoma produced according to <1> to <12> The antibody, antibody fragment thereof, fusion antibody or fusion antibody fragment thereof according to any one of the above.

[0143] <14> A nucleic acid comprising a coding sequence according to <1> to <12> The nucleotide sequence of any one of the antibodies, antibody fragments thereof, fusion antibodies or fusion antibody fragments thereof.

[0144] <15> A transformed cell comprising <14> The nucleic acid vector.

[0145] <16> A method for producing <1> to <12> A method for producing an antibody, an antibody fragment thereof, a fusion antibody or a fusion antibody fragment thereof according to any one of the preceding claims, the method comprising:

[0146] Training basis <13> The hybridoma or <15> the transformant cell, and

[0147] Collect the culture solution according to <1> to <12> The antibody, antibody fragment thereof, fusion antibody or fusion antibody fragment thereof according to any one of the above.

[0148] <17> A composition comprising <1> to <12> The antibody, antibody fragment thereof, fusion antibody or fusion antibody fragment thereof according to any one of the above.

[0149] <18> according to <17> The composition is a composition for detecting or measuring antigens present in the brain.

[0150] <19> according to <17> The composition is a composition for diagnosing or treating brain diseases.

[0151] <20> A method for detecting or measuring an antigen present in the brain, the method using <1> to <12> The antibody, antibody fragment, fusion antibody or fusion antibody fragment thereof according to any one of <17> The composition.

[0152] <21> A method for diagnosing or treating a brain disease, the method comprising: <1> to <12> The antibody, antibody fragment, fusion antibody or fusion antibody fragment thereof according to any one of <17> The composition.

[0153] <22> A method for improving the property of an antibody, an antibody fragment thereof, a fusion antibody or a fusion antibody fragment thereof to accumulate in the brain, the method using <1> to <12> The antibody, antibody fragment, fusion antibody or fusion antibody fragment thereof according to any one of <17> The composition.

[0154] <23> A method for increasing the amount of an antibody, an antibody fragment thereof, a fusion antibody or a fusion antibody fragment thereof in the brain, the method using a method according to <1> to <12> The antibody, antibody fragment, fusion antibody or fusion antibody fragment thereof according to any one of <17> The composition.

[0155] Advantageous Effects of the Invention

[0156] The CADM3 binding molecules of the present invention not only improve the property of the binding molecule itself to accumulate in the brain by specifically binding to CADM3, but can also be used to treat brain diseases by modifying the CADM3 binding molecule with another target molecule and transporting and retaining the target molecule in the brain. As specific CADM3 binding molecules of the present invention, antibodies or antibody fragments thereof are exemplified. The antibodies or antibody fragments thereof of the present invention are antibodies or antibody fragments thereof that have the property of accumulating in the brain by binding to CADM3 in the brain. Therefore, the antibodies or antibody fragments thereof of the present invention can be used as compositions for detecting or measuring antigens (CADM3, or CADM3 and another antigen present in the brain) present in the brain, compositions for diagnosing brain diseases, and pharmaceutical compositions for treating brain diseases. Sequence Listing <110> Kyowa Hakko Kirin Co., Ltd. Kagoshima University, a national university corporation <120> Antibodies that bind to cell adhesion molecule 3 <130> W527352 <150> JP2018-120477 <151> 2018-06-26 <160> 178 <170> PatentIn version 3.5 <210> 1 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L5 excluding signal sequence <400> 1 caggtgcagc tcgtggagtc tgggggaggc ttggtgcagg ctggggggtc tctgagactc 60 tcctgtgcag cctctggaag catcgtcagt gtcaatgcca tgggctggta ccgccaggct 120 ccagggaagc agcgcgagtt ggtcgcaact attactagtg ggggtagcac aaactatgca 180 gactccgcga agggccgatt caccatctcc agagacaacg ccaagaacac gatgtatctg 240 caaatgaaca gcctgaaacc tgaggacaca gccgtctatt actgtaacgg ggaattctgg 300 tcgcgccggg acacacgccc cccaggggtc gtaaactact ggggccaggg gacccaggtc 360 accgtctcct ca 372 <210> 2 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L5 excluding the signal sequence <400> 2 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Val Ser Val Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Thr Ile Thr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Met Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Gly Glu Phe Trp Ser Arg Arg Asp Thr Arg Pro Pro Gly Val Val Asn 100 105 110 Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 3 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR1 of iCADM3_3R1-L5 <400> 3 Val Asn Ala Met Gly 1 5 <210> 4 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR2 of iCADM3_3R1-L5 <400> 4 Thr Ile Thr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala Lys Gly 1 5 10 15 <210> 5 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR3 of iCADM3_3R1-L5 <400> 5 Glu Phe Trp Ser Arg Arg Asp Thr Arg Pro Pro Gly Val Val Asn Tyr 1 5 10 15 <210> 6 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L8 excluding signal sequence <400> 6 caggtgcagc tcgtggagtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60 tcctgtgcag cctctggacg caccttcagt aattatgccc ggggctggtt ccgccaggct 120 ccagggaagg agcgtgagtt tgtagcagct attgactaca gtggtggtag cacaaactat 180 gcagactccg cgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240 ctgcaaatga acagcctgaa acccggggac acggccgtttattactgtgc agcgcccgca 300 agccggcgtc ctagttggga tgctgatggg tatgactact ggggccaggg gacccaggtc 360 accgtctcct ca 372 <210> 7 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L8 excluding signal sequence <400> 7 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 Arg Thr Phe Ser Asn Tyr 20 25 30 Ala Arg Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45 Ala Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Gly Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 8 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR1 of iCADM3_3R1-L8 <400> 8 Asn Tyr Ala Arg Gly 1 5 <210> 9 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR2 of iCADM3_3R1-L8 <400> 9 Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala Lys 1 5 10 15 Gly <210> 10 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR3 of iCADM3_3R1-L8 <400> 10 Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp Tyr 1 5 10 15 <210> 11 <211> 363 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L10 excluding signal sequence <400> 11 caggtgcagc tcgtggagtc tgggggaggc ttggtgcagg ctggggggtc tctgagactc 60 tcctgtgcag cctctggaag catcttcagt atacatgcca tgggctggta ccgtcaggct 120 ccagggaagc agcgcgagtt ggtcgcaact gttactagtg gtggtagcac aaactatgca 180 gactccgtga agggccgatt caccatctcc agagacaacg ccaagaacac ggtgtatctg 240 caaatgaaca gcctgaaacc tgaggacaca gccgtctatt actgtaatgc agaaaccccc 300 tactatagta gtacttacta cacgaactac tggggccagg ggacccaggt caccgtctcc 360 tca 363 <210> 12 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L10 excluding the signal sequence <400> 12 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Ile His 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val 35 40 45 Ala Thr Val Thr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Thr Pro Tyr Tyr Ser Ser Thr Tyr Tyr Thr Asn Tyr Trp Gly 100 105 110 Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 13 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR1 of iCADM3_3R1-L10 <400> 13 Ile His Ala Met Gly 1 5 <210> 14 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR2 of iCADM3_3R1-L10 <400> 14 Thr Val Thr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 15 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR3 of iCADM3_3R1-L10 <400> 15 Glu Thr Pro Tyr Tyr Ser Ser Thr Tyr Tyr Thr Asn Tyr 1 5 10 <210> 16 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11 excluding signal sequence <400> 16 caggtgcagc tcgtggagtc tgggggaggc ttggtgcagg ctggggggtc tctgagactt 60 tcctgtgcag cctctggaag catcttcagc ttcaatgcca tgggctggta ccgccaggct 120 ccagggaagc agcgcgggtt ggtcgcagtt attactagtg gtggttacac aaactatgcg 180 gactccgtga agggccgatt caccatcacc agagacaacg ccaagaacac ggtgtatctg 240 caaatgaaca gcctgaaacc tgaggacaca gccgtctatt actgtaatgc agaaggagtc 300 tacagcgact atgtgatcat gaactactgg ggccagggga cccaggtcac cgtctcctca 360 <210> 17 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11 excluding the signal sequence <400> 17 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Gly Leu Val 35 40 45 Ala Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Thr Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 18 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR1 of iCADM3_3R1-L11 <400> 18 Phe Asn Ala Met Gly 1 5 <210> 19 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR2 of iCADM3_3R1-L11 <400> 19 Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 20 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of CDR3 of iCADM3_3R1-L11 <400> 20 Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr 1 5 10 <210> twenty one <211> 369 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of CADM301 VH excluding signal sequence <400> twenty one caggtgcagc tggtgcaatc tggggctgag gtgaggaggc ctgggacctc agtgaaagtc 60 tcctgcaagg cttctggata cagcttcacc agttatgata ttaactgggt gcgcctggcc 120 actggacaag ggcttgagtg gatggggtgg atgaacccta acactggtga tacaggctct 180 ccacagaagt tccaggacag agtcaccatg accagggaca tctccacagg cacagcctac 240 ttagaactga gaggcctgaa gtctgaggac acggccattt attattgtgc gagaggcttc 300 ctggtgacag catataccgc tgagttcttc ccgcactggg gccagggcac cctggtcacc 360 gtctcctca 369 <210> twenty two <211> 123 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM301 excluding signal sequence <400> twenty two Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Arg Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Asp Ile Asn Trp Val Arg Leu Ala Thr Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Met Asn Pro Asn Thr Gly Asp Thr Gly Ser Pro Gln Lys Phe 50 55 60 Gln Asp Arg Val Thr Met Thr Arg Asp Ile Ser Thr Gly Thr Ala Tyr 65 70 75 80 Leu Glu Leu Arg Gly Leu Lys Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Gly Phe Leu Val Thr Ala Tyr Thr Ala Glu Phe Phe Pro His 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> twenty three <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM301 <400> twenty three Ser Tyr Asp Ile Asn 1 5 <210> twenty four <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM301 <400> twenty four Trp Met Asn Pro Asn Thr Gly Asp Thr Gly Ser Pro Gln Lys Phe Gln 1 5 10 15 Asp <210> 25 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM301 <400> 25 Gly Phe Leu Val Thr Ala Tyr Thr Ala Glu Phe Phe Pro His 1 5 10 <210> 26 <211> 324 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VL of CADM301 excluding signal sequence <400> 26 tcctatgtgc tgactcagcc accctcagtg tcagtggccc caggaaagac ggccaggctc 60 acctgtgggg gaaacaacat tggaagtaaa agtgttcact ggtaccagca gaggccaggc 120 caggcccctg tgctggtcat aaattatgat agtgaccggc cctctggggat ccctgagcga 180 ttctctggct ccaactctga gaacacggcc accctgacca tcagcagggt cgaagccggg 240 gatgaggccg actattactg tcaggtgtgg gatagtggta gtgatcatgt ggtattcggc 300 ggaggaaccc agctgatcat ttta 324 <210> 27 <211> 108 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VL of CADM301 excluding signal sequence <400> 27 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Lys 1 5 10 15 Thr Ala Arg Leu Thr Cys Gly Gly Asn Asn Ile Gly Ser Lys Ser Val 20 25 30 His Trp Tyr Gln Gln Arg Pro Gly Gln Ala Pro Val Leu Val Ile Asn 35 40 45 Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Glu Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu Ala Gly 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Gly Ser Asp His 85 90 95 Val Val Phe Gly Gly Gly Thr Gln Leu Ile Ile Leu 100 105 <210> 28 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR1 of CADM301 <400> 28 Gly Gly Asn Asn Ile Gly Ser Lys Ser Val His 1 5 10 <210> 29 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR2 of CADM301 <400> 29 Tyr Asp Ser Asp Arg Pro Ser 1 5 <210> 30 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR3 of CADM301 <400> 30 Gln Val Trp Asp Ser Gly Ser Asp His Val Val 1 5 10 <210> 31 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3102 excluding signal sequence <400> 31 cagatgcagc tagtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggaggg atcatcccta tgtctggcac agcaaactac 180 gcacagaaat tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac acggccgtct actactgtgc gagagttgag 300 gaaagtggct ggtacgacca ctaccacggt atggacgtct ggggccaagg gaccacggtc 360 accgtctcct ca 372 <210> 32 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3102 excluding the signal sequence <400> 32 Gln Met 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 Ser 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 Ser Gly 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 Val Glu Glu Ser Gly Trp Tyr Asp His Tyr His Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 33 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3102 <400> 33 Ser Tyr Ala Ile Ser 1 5 <210> 34 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3102 <400> 34 Gly Ile Ile Pro Met Ser Gly Thr Ala Asn Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 35 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3102 <400> 35 Val Glu Glu Ser Gly Trp Tyr Asp His Tyr His Gly Met Asp Val 1 5 10 15 <210> 36 <211> 336 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VL of CADM3102 excluding signal sequence <400> 36 gatgttgtga tgactcagtc tccactctcc ctgcccgtca cccctggaga gccggcctcc 60 atctcctgca cgtctagtca gagcctcctg tatagtaatg gattcaacta tttggattgg 120 tacctgcaga aaccagggca gtctccacag ctcctgatct atttgggttc taatcgggcc 180 tccggggtcc ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc 240 agtagagtgg aggctgagga tgttggggtg tattactgca tgcaagctct aacaactcat 300 cccacttttg gcggagggac caaagtggat atcaaa 336 <210> 37 <211> 112 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VL of CADM3102 excluding signal sequence <400> 37 Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Thr Ser Ser Gln Ser Leu Leu Tyr Ser 20 25 30 Asn Gly Phe Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala 85 90 95 Leu Thr Thr His Pro Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys 100 105 110 <210> 38 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR1 of CADM3102 <400> 38 Thr Ser Ser Gln Ser Leu Leu Tyr Ser Asn Gly Phe Asn Tyr Leu Asp 1 5 10 15 <210> 39 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR2 of CADM3102 <400> 39 Leu Gly Ser Asn Arg Ala Ser 1 5 <210> 40 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR3 of CADM3102 <400> 40 Met Gln Ala Leu Thr Thr His Pro Thr 1 5 <210> 41 <211> 648 <212> DNA <213> Artificial sequence <220> <223> pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L5 VHH、pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L8 VHH、pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L5 VHH VHH is a self-directed video company <400> 41 cagtttgtgc tttctcagcc aaactctgtg tctacgaatc tcggaagcac agtcaaactg 60 tcttgcaagc gcagcactgg taacattgga agcaattatg tgagctggta ccagcagcat 120 gagggaagat ctcccaccac tatgattat agggatgata agagaccaga tggagttcct 180 gacaggttct ctggctccat tgacagatct tccgactcag ccctcctgac aatcaataat 240 gtgcagactg aagatgaagc tgactacttc tgtcagtctt acagtagtgg tattaatatt 300 ttcggcggtg gaaccaagct cactgtccta ggtcagccca aggccgcccc ctcggtcact 360 ctgttcccgc cctcctctga ggagcttcaa gccaacaagg ccacactggt gtgtctcata 420 agtgacttct acccgggagc cgtgacagtg gcctggaagg cagatagcag ccccgtcaag 480 gcgggagtgg agaccaccac accctccaaa caaagcaaca acaagtacgc ggccagcagc 540 tacctgagcc tgacgcctga gcagtggaag tcccacagaa gctacagctg ccaggtcacg 600 catgaaggga gcaccgtgga gaagacagtg gcccctacag aatgttca 648 <210> 42 <211> 216 <212> PRT <213> Artificial sequence <220> <223> Amino acid sequences of light chain antibodies excluding the signal sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L5 VHH, pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L8 VHH, and pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L10 VHH <400> 42 Gln Phe Val Leu Ser Gln Pro Asn Ser Val Ser Thr Asn Leu Gly Ser 1 5 10 15 Thr Val Lys Leu Ser Cys Lys Arg Ser Thr Gly Asn Ile Gly Ser Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln His Glu Gly Arg Ser Pro Thr Thr Met 35 40 45 Ile Tyr Arg Asp Asp Lys Arg Pro Asp Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Ile Asp Arg Ser Ser Asp Ser Ala Leu Leu Thr Ile Asn Asn 65 70 75 80 Val Gln Thr Glu Asp Glu Ala Asp Tyr Phe Cys Gln Ser Tyr Ser Ser 85 90 95 Gly Ile Asn Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 43 <211> 1764 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Base sequence of heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L5 VHH excluding signal sequence <400> 43 gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggagaatc cctgaaactc 60 tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120 ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180 cgcgactccg tgaagggccg attcactatc tccagagatg atgcaaaaaa caccctatac 240 ctgcaaatgg acagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300 aattatatat attatgggtc cttctttgat tactggggcc aaggatcat ggtcacagtc 360 tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420 tccgagagca cagccgccct gggctgcctg gtcaaggact acttccccga accggtgacg 480 gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttccccggc tgtcctacag 540 tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600 aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660 gagtccaaat atggtcccccc 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 aggtgcagct cgtggagtct gggggaggct tggtgcaggc tggggggctct 1440 ctgagactct cctgtgcagc ctctggaagc atcgtcagtg tcaatgccat gggctggtac 1500 cgccaggctc cagggaagca gcgcgagttg gtcgcaacta ttactagtgg gggtagcaca 1560 aactatgcag actccgcgaa gggccgattc accatctcca gagacaacgc caagaacacg 1620 atgtatctgc aaatgaacag cctgaaacct gaggacacag ccgtctatta ctgtaacggg 1680 gaattctggt cgcgccggga cacacgcccc ccaggggtcg taaactactg gggccagggg 1740 acccaggtca ccgtctcctc atga 1764 <210> 44 <211> 587 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L5 VHH excluding the signal sequence <400> 44 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 Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 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 Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser 465 470 475 480 Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Val Ser Val Asn Ala 485 490 495 Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala 500 505 510 Thr Ile Thr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala Lys Gly 515 520 525 Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Met Tyr Leu Gln 530 535 540 Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Gly 545 550 555 560 Glu Phe Trp Ser Arg Arg Asp Thr Arg Pro Pro Gly Val Val Asn Tyr 565 570 575 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 580 585 <210> 45 <211> 1764 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Base sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L8 VHH excluding the signal sequence <400> 45 gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60 tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120 ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180 cgcgactccg tgaagggccg attcactatc tccagagatg atgcaaaaaa caccctatac 240 ctgcaaatgg acagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300 aattatatat attatgggtc cttctttgat tactggggcc aaggatcat ggtcacagtc 360 tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420 tccgagagca cagccgccct gggctgcctg gtcaaggact acttccccga accggtgacg 480 gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttccccggc tgtcctacag 540 tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600 aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660 gagtccaaat atggtcccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720 tcagtcttcc tgttcccccc aaaacccaag gacactctca tgatctcccg gacccctgag 780 gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840 gtggatggcg tggaggtgca taatgccaag acaaagccgc gggagagca 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 aggtgcagct cgtggagtct gggggaggct tggtgcagcc tggggggctct 1440 ctgagactct cctgtgcagc ctctggacgc accttcagta attatgcccg gggctggttc 1500 cgccaggctc cagggaagga gcgtgagttt gtagcagcta ttgactacag tggtggtagc 1560 acaaactatg cagactccgc gaagggccga ttcaccatct ccagagacaa cgccaagaac 1620 acggtgtatc tgcaaatgaa cagcctgaaa cccggggaca cggccgttta ttactgtgca 1680 gcgcccgcaa gccggcgtcc tagttggggat gctgatgggt atgactactg gggccagggg 1740 acccaggtca ccgtctcctc atga 1764 <210> 46 <211> 587 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L8 VHH excluding the signal sequence <400> 46 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 Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 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 Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser 465 470 475 480 Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Asn Tyr Ala 485 490 495 Arg Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val Ala 500 505 510 Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala Lys 515 520 525 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 530 535 540 Gln Met Asn Ser Leu Lys Pro Gly Asp Thr Ala Val Tyr Tyr Cys Ala 545 550 555 560 Ala Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp Tyr 565 570 575 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 580 585 <210> 47 <211> 1755 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L10VHH excluding the signal sequence <400> 47 gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60 tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120 ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180 cgcgactccg tgaagggccg attcactatc tccagagatg atgcaaaaaa caccctatac 240 ctgcaaatgg acagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300 aattatatat attatgggtc 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 cccagcaca ccaggtgga caagagagtt 660 gagtccaaat atggtccccc atgcccacca tgcccagcac ctgagttcga ggggggacca 720 tcagtcttcc tgttcccccc aaaacccaag gatactca tgatctcccg gacccctgag 780 gtcacgtgcg tggtggtgga cgtgagccag gaagaccccg aggtccagtt caactggtac 840 gtggatggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gttcaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa cggcaggag 960 tacaagtgca aggtctccaa caaggccctc ccgtcctcca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga gccacaggtg tacaccctgc cccatcca ggaggagatg 1080 accagaacc aggtcagcct gacctgcctg gtcaaggct tctacccg cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacactaca 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 aggtgcagct cgtggagtct gggggaggct tggtgcaggc tggggggtct 1440 ctgagactct cctgtgcagc ctctggaagc atcttcagta tacatgccat gggctggtac 1500 cgtcaggctc cagggaagca gcgcgagttg gtcgcaactg ttactagtgg tggtagcaca 1560 aactatgcag actccgtgaa gggccgattc accatctcca gagacaacgc caagaacacg 1620 gtgtatctgc aaatgaacag cctgaaacct gaggacacag ccgtctatta ctgtaatgca 1680 gaaaccccct actatagtag tacttactac acgaactact ggggccaggg gacccaggtc 1740 accgtctcct catga 1755 <210> 48 <211> 584 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L10VHH excluding the signal sequence <400> 48 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 Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 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 Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser 465 470 475 480 Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Ile His Ala 485 490 495 Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala 500 505 510 Thr Val Thr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys Gly 515 520 525 Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln 530 535 540 Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala 545 550 555 560 Glu Thr Pro Tyr Tyr Ser Ser Thr Tyr Tyr Thr Asn Tyr Trp Gly Gln 565 570 575 Gly Thr Gln Val Thr Val Ser Ser 580 <210> 49 <211> 1752 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Base sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L11VHH excluding the signal sequence <400> 49 gaggtgcagc tggtggaatc tgggggaggc ttagtgcagc ctggaagatc cctgaaactc 60 tcctgtgcag cctcaggatt cactttcagt aactatgcca tggcttgggt ccgccgggct 120 ccaacgaagg gtctggagtg ggtcgcatcc attagtaatg gtggtggtaa cacttactat 180 cgcgactccg tgaagggccg attcactatc tccagagatg atgcaaaaaa caccctatac 240 ctgcaaatgg acagtctgag gtctgaggac acggccactt attactgtgc aagacacggg 300 aattatatat attatgggtc cttctttgat tactggggcc aaggatcat ggtcacagtc 360 tcctcagcta gcaccaaggg gccatccgtc ttccccctgg cgccctgctc caggagcacc 420 tccgagagca cagccgccct gggctgcctg gtcaaggact acttccccga accggtgacg 480 gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttccccggc tgtcctacag 540 tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacg 600 aagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagagagtt 660 gagtccaaat atggtcccccc 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 aggtgcagct cgtggagtct gggggaggct tggtgcaggc tggggggctct 1440 ctgagacttt cctgtgcagc ctctggaagc atcttcagct tcaatgccat gggctggtac 1500 cgccaggctc cagggaagca gcgcgggttg gtcgcagtta ttactagtgg tggttacaca 1560 aactatgcgg actccgtgaa gggccgattc accatcacca gagacaacgc caagaacacg 1620 gtgtatctgc aaatgaacag cctgaaacct gaggacacag ccgtctatta ctgtaatgca 1680 gaaggagtct acagcgacta tgtgatcatg aactactggg gccaggggac ccaggtcacc 1740 gtctcctcat ga 1752 <210> 50 <211> 583 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of the heavy chain antibody sequence of pCI_AVM-hLG4PE(R409K)-iCADM3_3R1-L11VHH excluding the signal sequence <400> 50 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 Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 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 Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser 465 470 475 480 Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Phe Asn Ala 485 490 495 Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Gly Leu Val Ala 500 505 510 Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys Gly 515 520 525 Arg Phe Thr Ile Thr Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln 530 535 540 Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala 545 550 555 560 Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln Gly 565 570 575 Thr Gln Val Thr Val Ser Ser 580 <210> 51 <211> 1197 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of human CADM3 including signal sequence <400> 51 atgggggccc cagccgcctc gctcctgctc ctgctcctgc tgttcgcctg ctgctgggcg 60 cccggcgggg ccaacctctc ccaggacgac agccagccct ggacatctga tgaaacagtg 120 gtggctggtg gcaccgtggt gctcaagtgc caagtgaaag atcacgagga ctcatccctg 180 caatggtcta accctgctca gcagactctc tactttgggg agaagagagc ccttcgagat 240 aatcgaattc agctggttac ctctacgccc cacgagctca gcatcagcat cagcaatgtg 300 gccctggcag acgagggcga gtacacctgc tcaatcttca ctatgcctgt gcgaactgcc 360 aagtccctcg tcactgtgct aggaattcca cagaagccca tcatcactgg tttaaatct 420 tcattacggg aaaaagacac agccacccta aactgtcagt cttctggggag caagcctgca 480 gcccggctca cctggagaaa gggtgaccaa gaactccacg gagaaccaac ccgcatacag gaagatccca atggtaaaac cttcactgtc agcagctcgg tgacattcca ggttacccgg gggatgatg gggcgagcat cgtgtgctct gtgaaccatg aatctctaaa gggagctgac agatccacct ctcaacgcat tgaagtttta tacacaccaa ctgcgatgat taggccagac cctccccatc ctcgtgaggg ccagaagctg ttgctacact gtgagggtcg cggcaatcca 780. gtcccccagc agtacctatg ggagaaggag ggcagtgtgc cacccctgaa gatgacccag 840 900. gagagtgcc tgatcttccc tttcctcaac aagagtgaca gtggcaccta cggctgcaca gccaccagca acatgggcag ctacaaggcc tactacaccc tcaatgttaa tgaccccagt ccggtgccct cctcctccag cacctaccac gccatcatcg gtgggatcgt ggctttcatt 1020 gtcttcctgc tgctcatcat gctcatcttc cttggccact acttgatccg gcacaaagga 1080 acctacctga cacatgaggc aaaaggctcc gacgatgctc cagacgcgga cacggccatc 1140 atcaatgcag aaggcgggca gtcaggaggg gacgacaaga aggaatattt catctag 1197 <210> 52 <211> 398 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of human CADM3 including signal sequence <400> 52 Met Gly Ala Pro Ala Ala Ser Leu Leu Leu Leu Leu Leu Leu Phe Ala 1 5 10 15 Cys Cys Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln 20 25 30 Pro Trp Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu 35 40 45 Lys Cys Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn 50 55 60 Pro Ala Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp 65 70 75 80 Asn Arg Ile Gln Leu Val Thr Ser Thr Pro His Glu Leu Ser Ile Ser 85 90 95 Ile Ser Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile 100 105 110 Phe Thr Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly 115 120 125 Ile Pro Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu 130 135 140 Lys Asp Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala 145 150 155 160 Ala Arg Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Glu Pro 165 170 175 Thr Arg Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser 180 185 190 Ser Val Thr Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Ser Ile Val 195 200 205 Cys Ser Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser 210 215 220 Gln Arg Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Asp 225 230 235 240 Pro Pro His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly 245 250 255 Arg Gly Asn Pro Val Pro Gln Gln Tyr Leu Trp Glu Lys Glu Gly Ser 260 265 270 Val Pro Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe 275 280 285 Leu Asn Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn 290 295 300 Met Gly Ser Tyr Lys Ala Tyr Tyr Thr Leu Asn Val Asn Asp Pro Ser 305 310 315 320 Pro Val Pro Ser Ser Ser Ser Thr Tyr His Ala Ile Ile Gly Gly Ile 325 330 335 Val Ala Phe Ile Val Phe Leu Leu Leu Ile Met Leu Ile Phe Leu Gly 340 345 350 His Tyr Leu Ile Arg His Lys Gly Thr Tyr Leu Thr His Glu Ala Lys 355 360 365 Gly Ser Asp Asp Ala Pro Asp Ala Asp Thr Ala Ile Ile Asn Ala Glu 370 375 380 Gly Gly Gln Ser Gly Gly Asp Asp Lys Lys Glu Tyr Phe Ile 385 390 395 <210> 53 <211> 1191 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of mouse CADM3 including signal sequence <400> 53 atgggggccc cttccgccct gcccctgctc ctgctcctcg cctgctcctg ggcgcccggc 60 ggggccaatc tttcccagga cgatagccag ccctggacat ctgatgaaac agttgtggct 120 ggtggcacag tggttctcaa gtgtcaagta aaagaccatg aagactcatc tctgcagtgg 180 tctaaccctg ctcagcagac cctatacttc ggggagaaga gagcccttcg agataatcgg 240 attcagctgg ttagctctac tccccatgag ctcagcatca gcatcagcaa tgtggcgctg 300 gccgatgagg gggagtacac gtgctccatc ttcactatgc ctgtgcgaac cgccaagtcc 360 cttgtcactg tgctcggaat cccacagaaa cccataatca ctggttataa gtcatcattg 420 cgggaaaagg agacagccac tctaaattgt cagtcttctg ggagcaaacc tgcagcccag 480 ctcacctgga ggaaaggtga ccaagaactc cacggggacc aaacacgaat ccaggaagat 540 cccaacggga aaaccttcac tgtgagcagc tcagtgtcat tccaggttac ccgggaggat 600 gatggagcaa acatcgtgtg ctctgtgaac catgaatctc tgaagggagc cgacagatcc 660 acttctcagc gcattgaagt gttatacaca ccaacagcca tgattaggcc agaacctgct 720 catcctcgag aaggccagaa gctgttgtta cattgtgagg ggcgtggcaa tccagtcccc 780 cagcagtacg tgtgggtaaa ggaaggcagt gagccacccc tcaagatgac ccaagagagt 840 gctctcatct tccccttttt gaataagagt gacagtggca cttatggctg tacagccaca 900 agcaacatgg gcagctatac agcctacttc accctcaatg tcaacgaccc cagtccagtg 960 ccctcgtcct ccagtaccta ccacgccatc attggaggga ttgtggcttt cattgtcttc 1020 ctgctgctca ttctgctcat tttccttgga cactatttga tccggcacaa aggaacctac 1080 ctgacacacg aagcgaaggg ttccgacgat gctccagatg cggatacggc catcatcaac 1140 gcagaaggcg ggcagtcagg cggggatgac aagaaggaat atttcatcta g 1191 <210> 54 <211> 396 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of mouse CADM3 including signal sequence <400> 54 Met Gly Ala Pro Ser Ala Leu Pro Leu Leu Leu Leu Leu Ala Cys Ser 1 5 10 15 Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln Pro Trp 20 25 30 Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu Lys Cys 35 40 45 Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn Pro Ala 50 55 60 Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp Asn Arg 65 70 75 80 Ile Gln Leu Val Ser Ser Thr Pro His Glu Leu Ser Ile Ser Ile Ser 85 90 95 Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile Phe Thr 100 105 110 Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly Ile Pro 115 120 125 Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu Lys Glu 130 135 140 Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala Ala Gln 145 150 155 160 Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Asp Gln Thr Arg 165 170 175 Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser Ser Val 180 185 190 Ser Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Asn Ile Val Cys Ser 195 200 205 Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser Gln Arg 210 215 220 Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Glu Pro Ala 225 230 235 240 His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly Arg Gly 245 250 255 Asn Pro Val Pro Gln Gln Tyr Val Trp Val Lys Glu Gly Ser Glu Pro 260 265 270 Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe Leu Asn 275 280 285 Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn Met Gly 290 295 300 Ser Tyr Thr Ala Tyr Phe Thr Leu Asn Val Asn Asp Pro Ser Pro Val 305 310 315 320 Pro Ser Ser Ser Ser Thr Tyr His Ala Ile Ile Gly Gly Ile Val Ala 325 330 335 Phe Ile Val Phe Leu Leu Leu Ile Leu Leu Ile Phe Leu Gly His Tyr 340 345 350 Leu Ile Arg His Lys Gly Thr Tyr Leu Thr His Glu Ala Lys Gly Ser 355 360 365 Asp Asp Ala Pro Asp Ala Asp Thr Ala Ile Ile Asn Ala Glu Gly Gly 370 375 380 Gln Ser Gly Gly Asp Asp Lys Lys Glu Tyr Phe Ile 385 390 395 <210> 55 <211> 1191 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of cynomolgus monkey CADM3 including signal sequence <400> 55 atgggggccc cagtcgcctt gctcctgctc ctgctgttcg cctgctgctg ggcgcccagt 60 ggggccaacc tctcccagga cgacagccag ccctggacat ctgatgaaac agtggtggct 120 ggtggcaccg tggtgctcaa gtgccaagtg aaagatcacg aggactcatc cctgcaatgg 180 tctaaccctg ctcagcagac tctctacttt ggggagaaga gagcccttcg agataatcga 240 attcagctgg ttacctctac tccccacgag ctcagcatca gcatcagcaa tgtggccctg 300 gcagacgagg gcgagtacac ctgctcaatc ttcactatgc ctgtacgaac tgccaagtcc 360 ctcgtcactg tgctaggaat tccacagaag cccatcatca ctggttataa atcttcatta 420 cgggaaaagg acacagccac cctaaactgt cagtcttctg ggagcaagcc tgcagcccgg 480 ctcacctgga gaaagggtga ccaagaactc cacggagaac caactcgcat acaggaagat 540 cccaatggta aaaccttcac tgtcagcagc tcggtgacat tccaggttac ccgggaggat 600 gatggggcga acatcgtgtg ctctgtgaac catgaatctc taaagggagc tgacagatcc 660 acctctcaac gcattgaagt tttatacaca ccgactgcga tgattaggcc agaccctccc 720 catcctcgtg agggccagaa gctgttgcta cactgtgagg gtcgtggcaa tccagtcccc 780 cagcagtacc tatgggagaa ggagggcagt gtgccacccc tgaagatgac ccaagagagt 840 gccctgatct tccccttcct caacaagagt gacagcggca cctacggctg cacggccacc 900 agcaacatgg gcagctacaa ggcctactac actctcaacg ttaatgaccc cagtccggtg 960 ccctcctcct ccagcaccta ccacgccatc atcggcggga tcgtggcttt cattgtcttc 1020 ctgctgctca tcatgctcat cttccttgga cattacttga tccggcacaa aggaacctac 1080 ctgacacatg aggcgaaagg ctccgacgat gccccagatg cggacacggc catcatcaat 1140 gcagaaggcg ggcagtcggg aggggacgac aagaaggaat atttcatcta g 1191 <210> 56 <211> 396 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of cynomolgus monkey CADM3 including signal sequence <400> 56 Met Gly Ala Pro Val Ala Leu Leu Leu Leu Leu Leu Phe Ala Cys Cys 1 5 10 15 Trp Ala Pro Ser Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln Pro Trp 20 25 30 Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu Lys Cys 35 40 45 Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn Pro Ala 50 55 60 Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp Asn Arg 65 70 75 80 Ile Gln Leu Val Thr Ser Thr Pro His Glu Leu Ser Ile Ser Ile Ser 85 90 95 Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile Phe Thr 100 105 110 Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly Ile Pro 115 120 125 Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu Lys Asp 130 135 140 Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala Ala Arg 145 150 155 160 Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Glu Pro Thr Arg 165 170 175 Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser Ser Val 180 185 190 Thr Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Asn Ile Val Cys Ser 195 200 205 Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser Gln Arg 210 215 220 Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Asp Pro Pro 225 230 235 240 His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly Arg Gly 245 250 255 Asn Pro Val Pro Gln Gln Tyr Leu Trp Glu Lys Glu Gly Ser Val Pro 260 265 270 Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe Leu Asn 275 280 285 Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn Met Gly 290 295 300 Ser Tyr Lys Ala Tyr Tyr Thr Leu Asn Val Asn Asp Pro Ser Pro Val 305 310 315 320 Pro Ser Ser Ser Ser Thr Tyr His Ala Ile Ile Gly Gly Ile Val Ala 325 330 335 Phe Ile Val Phe Leu Leu Leu Ile Met Leu Ile Phe Leu Gly His Tyr 340 345 350 Leu Ile Arg His Lys Gly Thr Tyr Leu Thr His Glu Ala Lys Gly Ser 355 360 365 Asp Asp Ala Pro Asp Ala Asp Thr Ala Ile Ile Asn Ala Glu Gly Gly 370 375 380 Gln Ser Gly Gly Asp Asp Lys Lys Glu Tyr Phe Ile 385 390 395 <210> 57 <211> 1713 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of hCADM3-FLAG_Fc including signal sequence <400> 57 atgggggccc cagccgcctc gctcctgctc ctgctcctgc tgttcgcctg ctgctgggcg 60 cccggcgggg ccaacctctc ccaggacgac agccagccct ggacatctga tgaaacagtg 120 gtggctggtg gcaccgtggt gctcaagtgc caagtgaaag atcacgagga ctcatccctg 180 caatggtcta accctgctca gcagactctc tactttgggg agaagagagc ccttcgagat 240 aatcgaattc agctggttac ctctacgccc cacgagctca gcatcagcat cagcaatgtg 300 gccctggcag acgagggcga gtacacctgc tcaatcttca ctatgcctgt gcgaactgcc 360 aagtccctcg tcactgtgct aggaattcca cagaagccca tcatcactgg ttataaatct 420 tcattacggg aaaaagacac agccacccta aactgtcagt cttctgggag caagcctgca 480 gcccggctca cctggagaaa gggtgaccaa gaactccacg gagaaccaac ccgcatacag 540 gaagatccca atggtaaaac cttcactgtc agcagctcgg tgacattcca ggttacccgg 600 gaggatgatg gggcgagcat cgtgtgctct gtgaaccatg aatctctaaa gggagctgac 660 agatccacct ctcaacgcat tgaagtttta tacacaccaa ctgcgatgat taggccagac 720 cctccccatc ctcgtgaggg ccagaagctg ttgctacact gtgagggtcg cggcaatcca 780. gtcccccagc agtacctatg ggagaaggag ggcagtgtgc cacccctgaa gatgacccag 840 900. gagagtgcc tgatcttccc tttcctcaac aagagtgaca gtggcaccta cggctgcaca gccaccagca acatgggcag ctacaaggcc tactacaccc tcaatgttaa tgaccccagt ccggtgccct cctcctccag cacctaccac tctagagcag actacaagga cgacgatgac aagactagtg acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 1140 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc agcactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggatgag 1440 ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 1500 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1560 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1620 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1680 cagaagagcc tctccctgtc tccgggtaaa tga 1713 <210> 58 <211> 570 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of hCADM3-FLAG_Fc including signal sequence <400> 58 Met Gly Ala Pro Ala Ala Ser Leu Leu Leu Leu Leu Leu Leu Phe Ala 1 5 10 15 Cys Cys Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln 20 25 30 Pro Trp Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu 35 40 45 Lys Cys Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn 50 55 60 Pro Ala Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp 65 70 75 80 Asn Arg Ile Gln Leu Val Thr Ser Thr Pro His Glu Leu Ser Ile Ser 85 90 95 Ile Ser Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile 100 105 110 Phe Thr Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly 115 120 125 Ile Pro Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu 130 135 140 Lys Asp Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala 145 150 155 160 Ala Arg Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Glu Pro 165 170 175 Thr Arg Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser 180 185 190 Ser Val Thr Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Ser Ile Val 195 200 205 Cys Ser Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser 210 215 220 Gln Arg Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Asp 225 230 235 240 Pro Pro His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly 245 250 255 Arg Gly Asn Pro Val Pro Gln Gln Tyr Leu Trp Glu Lys Glu Gly Ser 260 265 270 Val Pro Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe 275 280 285 Leu Asn Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn 290 295 300 Met Gly Ser Tyr Lys Ala Tyr Tyr Thr Leu Asn Val Asn Asp Pro Ser 305 310 315 320 Pro Val Pro Ser Ser Ser Ser Thr Tyr His Ser Arg Ala Asp Tyr Lys 325 330 335 Asp Asp Asp Asp Lys Thr Ser Asp Lys Thr His Thr Cys Pro Pro Cys 340 345 350 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 355 360 365 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 370 375 380 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 385 390 395 400 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 405 410 415 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 420 425 430 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 435 440 445 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 450 455 460 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 465 470 475 480 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 485 490 495 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 500 505 510 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 515 520 525 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 530 535 540 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 545 550 555 560 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 565 570 <210> 59 <211> 1707 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of mCADM3-FLAG_Fc including signal sequence <400> 59 atgggggccc cttccgccct gcccctgctc ctgctcctcg cctgctcctg ggcgcccggc 60 ggggccaatc tttcccagga cgatagccag ccctggacat ctgatgaaac agttgtggct 120 ggtggcacag tggttctcaa gtgtcaagta aaagaccatg aagactcatc tctgcagtgg 180 tctaaccctg ctcagcagac cctatacttc ggggagaaga gagcccttcg agataatcgg 240 attcagctgg ttagctctac tccccatgag ctcagcatca gcatcagcaa tgtggcgctg 300 gccgatgagg gggagtacac gtgctccatc ttcactatgc ctgtgcgaac cgccaagtcc 360 cttgtcactg tgctcggaat cccacagaaa cccataatca ctggttataa gtcatcattg 420 cgggaaaagg agacagccac tctaaattgt cagtcttctg ggagcaaacc tgcagcccag 480 ctcacctgga ggaaaggtga ccaagaactc cacggggacc aaacacgaat ccaggaagat 540 cccaacggga aaaccttcac tgtgagcagc tcagtgtcat tccaggttac ccgggaggat 600 gatggagcaa acatcgtgtg ctctgtgaac catgaatctc tgaagggagc cgacagatcc 660 acttctcagc gcattgaagt gttatacaca ccaacagcca tgattaggcc agaacctgct 720 catcctcgag aaggccagaa gctgttgtta cattgtgagg ggcgtggcaa tccagtcccc 780 cagcagtacg tgtgggtaaa ggaaggcagt gagccacccc tcaagatgac ccaagagagt 840 gctctcatct tccccttttt gaataagagt gacagtggca cttatggctg tacagccaca 900 agcaacatgg gcagctatac agcctacttc accctcaatg tcaacgaccc cagtccagtg 960 ccctcgtcct ccagtaccta ccactctaga gcagactaca aggacgacga tgacaagact 1020 agtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1080 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1140 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1200 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1260 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1320 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1380 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 1440 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1500 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1560 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1620 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1680 agcctctccc tgtctccggg taaatga 1707 <210> 60 <211> 568​​​​​​​​​​​Met Gly Ala Pro Ser Ala Leu Pro Leu Leu Leu Leu Leu Ala Cys Ser 1 5 10 15 Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln Pro Trp 20 25 30 Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu Lys Cys 35 40 45 Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn Pro Ala 50 55 60 Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp Asn Arg 65 70 75 80 Ile Gln Leu Val Ser Ser Thr Pro His Glu Leu Ser Ile Ser Ile Ser 85 90 95 Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile Phe Thr 100 105 110 Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly Ile Pro 115 120 125 Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu Lys Glu 130 135 140 Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala Ala Gln 145 150 155 160 Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Asp Gln Thr Arg 165 170 175 Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser Ser Val 180 185 190 Ser Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Asn Ile Val Cys Ser 195 200 205 Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser Gln Arg 210 215 220 Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Glu Pro Ala 225 230 235 240 His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly Arg Gly 245 250 255 Asn Pro Val Pro Gln Gln Tyr Val Trp Val Lys Glu Gly Ser Glu Pro 260 265 270 Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe Leu Asn 275 280 285 Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn Met Gly 290 295 300 Ser Tyr Thr Ala Tyr Phe Thr Leu Asn Val Asn Asp Pro Ser Pro Val 305 310 315 320 Pro Ser Ser Ser Ser Thr Tyr His Ser Arg Ala Asp Tyr Lys Asp Asp 325 330 335 Asp Asp Lys Thr Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 340 345 350 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 355 360 365 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 370 375 380 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 385 390 395 400 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 405 410 415 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 420 425 430 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 435 440 445 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 450 455 460 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 465 470 475 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 485 490 495 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 500 505 510 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 515 520 525 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 530 535 540 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 545 550 555 560 Ser Leu Ser Leu Ser Pro Gly Lys 565 <210> 61 <211> 1707 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of rCADM3-FLAG_Fc including signal sequence <400> 61 atgggggccc cttccgccct gcccctgctc ctgctcctcg cctgctcctg ggcgcccggc 60 ggggccaatc tttcccagga cgatagccag ccctggacgt ctgatgaaac agtggtggct 120 ggtggcacag tagtgctcaa gtgccaagtg aaagaccatg aagactcatc tctgcagtgg 180 tctaaccctg cccagcagac tctatacttt ggggagaaaa gagcccttcg agataatcgg 240 attcagctgg ttagctccac cccgcatgag ctcagcatca gcatcagcaa cgtggcactg 300 gccgacgagg gcgagtacac atgctccatc ttcactatgc ctgtgcggac cgccaagtcc 360 ctcgtcactg tgctcggaat cccacagaaa cccataatca ctggttataa gtcatcgttg 420 cgggaaaagg agacagccac tctaaattgt cagtcttctg ggagcaaacc tgcagcccag 480 ctcgcctgga gaaaaggtga ccaagaactc cacggggacc agacgcgaat ccaggaagat 540 cccaatggga aaaccttcac tgtgagcagc tcggtgtcat tccaggttac ccgggatgat 600 gatggagcaa acgtcgtgtg ctctgtgaac catgaatctc tgaagggagc tgacagatcc 660 acctctcagc gcattgaagt gttatacaca ccaacagcca tgattaggcc agaacctgct 720 catcctcgtg aaggccagaa gctgttgtta cattgtgagg ggcgtggcaa tccagtccct 780 cagcagtacg tgtgggtaaa agaaggcagc gagccacccc tcaagatgac ccaagagagt 840 gcactcatct tcccattttt gaacaaaagt gacagtggca cctatggctg tacagccacg 900 agcaacatgg gcagctatac agcctacttc actctcaatg tcaacgaccc tagtccagtg 960 ccctcatcct ccagtactta ccactctaga gcagactaca aggaggacga tgacaagact 1020 agtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1080 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1140 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1200 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1260 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1320 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1380 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 1440 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1500 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1560 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1620 gggacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1680 agcctctccc tgtctccggg taaatga 1707 <210> 62 <211> 568 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of rCADM3-FLAG_Fc including signal sequence <400> 62 Met Gly Ala Pro Ser Ala Leu Pro Leu Leu Leu Leu Leu Ala Cys Ser 1 5 10 15 Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln Pro Trp 20 25 30 Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu Lys Cys 35 40 45 Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn Pro Ala 50 55 60 Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp Asn Arg 65 70 75 80 Ile Gln Leu Val Ser Ser Thr Pro His Glu Leu Ser Ile Ser Ile Ser 85 90 95 Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile Phe Thr 100 105 110 Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly Ile Pro 115 120 125 Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu Lys Glu 130 135 140 Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala Ala Gln 145 150 155 160 Leu Ala Trp Arg Lys Gly Asp Gln Glu Leu His Gly Asp Gln Thr Arg 165 170 175 Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser Ser Val 180 185 190 Ser Phe Gln Val Thr Arg Asp Asp Asp Gly Ala Asn Val Val Cys Ser 195 200 205 Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser Gln Arg 210 215 220 Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Glu Pro Ala 225 230 235 240 His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly Arg Gly 245 250 255 Asn Pro Val Pro Gln Gln Tyr Val Trp Val Lys Glu Gly Ser Glu Pro 260 265 270 Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe Leu Asn 275 280 285 Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn Met Gly 290 295 300 Ser Tyr Thr Ala Tyr Phe Thr Leu Asn Val Asn Asp Pro Ser Pro Val 305 310 315 320 Pro Ser Ser Ser Ser Thr Tyr His Ser Arg Ala Asp Tyr Lys Asp Asp 325 330 335 Asp Asp Lys Thr Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 340 345 350 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 355 360 365 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 370 375 380 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 385 390 395 400 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 405 410 415 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 420 425 430 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 435 440 445 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 450 455 460 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 465 470 475 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 485 490 495 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 500 505 510 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 515 520 525 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 530 535 540 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 545 550 555 560 Ser Leu Ser Leu Ser Pro Gly Lys 565 <210> 63 <211> 1683 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Base sequence of hCADM3-GST including signal sequence <400> 63 atgggggccc cagccgcctc gctcctgctc ctgctcctgc tgttcgcctg ctgctgggcg 60 cccggcgggg ccaacctctc ccaggacgac agccagccct ggacatctga tgaaacagtg 120 gtggctggtg gcaccgtggt gctcaagtgc caagtgaaag atcacgagga ctcatccctg 180 caatggtcta accctgctca gcagactctc tactttgggg agaagagagc ccttcgagat 240 aatcgaattc agctggttac ctctacgccc cacgagctca gcatcagcat cagcaatgtg 300 gccctggcag acgagggcga gtacacctgc tcaatcttca ctatgcctgt gcgaactgcc 360 aagtccctcg tcactgtgct aggaattcca cagaagccca tcatcactgg ttataaatct 420 tcattacggg aaaaagacac agccacccta aactgtcagt cttctgggag caagcctgca 480 gcccggctca cctggagaaa gggtgaccaa gaactccacg gagaaccaac ccgcatacag 540 gaagatccca atggtaaaac cttcactgtc agcagctcgg tgacattcca ggttacccgg 600 gaggatgatg gggcgagcat cgtgtgctct gtgaaccatg aatctctaaa gggagctgac 660 agatccacct ctcaacgcat tgaagtttta tacacaccaa ctgcgatgat taggccagac 720 cctccccatc ctcgtgaggg ccagaagctg ttgctacact gtgagggtcg cggcaatcca 780 gtcccccagc agtacctatg ggaaggag ggcagtgtgc cacccctgaa gatgacccag 840 gagagtgccc tgatcttccc tttcctcaac aagagtgaca gtggcaccta cggctgcaca 900 gccaccagca acatgggcag ctacaaggcc tactacaccc tcaatgttaa tgaccccagt 960 ccggtgccct cctcctccag cacctaccac ggtaccctgg aagttctgtt ccaggggccc 1020 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt 1080 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa 1140 tggcgaaaca aaaagtttga attgggtttg gagttccca atcttcctta ttatattgat 1200 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac 1260 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg 1320 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt 1380 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa 1440 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat 1500 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa 1560 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca 1620 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat 1680 tga 1683 <210> 64 <211> 560 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of hCADM3-GST including signal sequence <400> 64 Met Gly Ala Pro Ala Ala Ser Leu Leu Leu Leu Leu Leu Leu Phe Ala 1 5 10 15 Cys Cys Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln 20 25 30 Pro Trp Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu 35 40 45 Lys Cys Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn 50 55 60 Pro Ala Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp 65 70 75 80 Asn Arg Ile Gln Leu Val Thr Ser Thr Pro His Glu Leu Ser Ile Ser 85 90 95 Ile Ser Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile 100 105 110 Phe Thr Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly 115 120 125 Ile Pro Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu 130 135 140 Lys Asp Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala 145 150 155 160 Ala Arg Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Glu Pro 165 170 175 Thr Arg Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser 180 185 190 Ser Val Thr Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Ser Ile Val 195 200 205 Cys Ser Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser 210 215 220 Gln Arg Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Asp 225 230 235 240 Pro Pro His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly 245 250 255 Arg Gly Asn Pro Val Pro Gln Gln Tyr Leu Trp Glu Lys Glu Gly Ser 260 265 270 Val Pro Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe 275 280 285 Leu Asn Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn 290 295 300 Met Gly Ser Tyr Lys Ala Tyr Tyr Thr Leu Asn Val Asn Asp Pro Ser 305 310 315 320 Pro Val Pro Ser Ser Ser Ser Thr Tyr His Gly Thr Leu Glu Val Leu 325 330 335 Phe Gln Gly Pro Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly 340 345 350 Leu Val Gln Pro Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr 355 360 365 Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys 370 375 380 Lys Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp 385 390 395 400 Gly Asp Val Lys Leu Thr Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala 405 410 415 Asp Lys His Asn Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile 420 425 430 Ser Met Leu Glu Gly Ala Val Leu Asp Ile Arg Tyr Gly Val Ser Arg 435 440 445 Ile Ala Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser 450 455 460 Lys Leu Pro Glu Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys 465 470 475 480 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr 485 490 495 Asp Ala Leu Asp Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala 500 505 510 Phe Pro Lys Leu Val Cys Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln 515 520 525 Ile Asp Lys Tyr Leu Lys Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln 530 535 540 Gly Trp Gln Ala Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp 545 550 555 560 <210> 65 <211> 1677 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of mCADM3-GST including signal sequence <400> 65 atgggggccc cttccgccct gcccctgctc ctgctcctcg cctgctcctg ggcgcccggc 60 ggggccaatc tttcccagga cgatagccag ccctggacat ctgatgaaac agttgtggct 120 ggtggcacag tggttctcaa gtgtcaagta aaagaccatg aagactcatc tctgcagtgg 180 tctaaccctg ctcagcagac cctatacttc ggggagaaga gagcccttcg agataatcgg 240 attcagctgg ttagctctac tccccatgag ctcagcatca gcatcagcaa tgtggcgctg 300 gccgatgagg gggagtacac gtgctccatc ttcactatgc ctgtgcgaac cgccaagtcc 360 cttgtcactg tgctcggaat cccacagaaa cccataatca ctggttataa gtcatcattg 420 cgggaaaagg agacagccac tctaaattgt cagtcttctg ggagcaaacc tgcagcccag 480 ctcacctgga ggaaaggtga ccaagaactc cacggggacc aaacacgaat ccaggaagat 540 cccaacggga aaaccttcac tgtgagcagc tcagtgtcat tccaggttac ccgggaggat 600 gatggagcaa acatcgtgtg ctctgtgaac catgaatctc tgaagggagc cgacagatcc 660 acttctcagc gcattgaagt gttatacaca ccaacagcca tgattaggcc agaacctgct 720 catcctcgag aaggccagaa gctgttgtta cattgtgagg ggcgtggcaa tccagtcccc 780 cagcagtacg tgtgggtaaa ggaaggcagt gagccacccc tcaagatgac ccaagagagt 840 gctctcatct tcccctttt gaataagagt gacagtggca cttatggctg tacagccaca 900 agcaacatgg gcagctatac agcctacttc accctcaatg tcaacgaccc cagtccagtg 960 cccgtcct ccagtaccta ccacggtacc ctggaagttc tgttccaggg gcccatgtcc 1020 cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa 1080 tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga 1140 aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat 1200 gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg 1260 ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt 1320 agatacggtg tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt 1380 cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat 1440 ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt 1500 ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt 1560 attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct 1620 ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggattga 1677 <210> 66 <211> 558 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of mCADM3-GST including signal sequence <400> 66 Met Gly Ala Pro Ser Ala Leu Pro Leu Leu Leu Leu Leu Ala Cys Ser 1 5 10 15 Trp Ala Pro Gly Gly Ala Asn Leu Ser Gln Asp Asp Ser Gln Pro Trp 20 25 30 Thr Ser Asp Glu Thr Val Val Ala Gly Gly Thr Val Val Leu Lys Cys 35 40 45 Gln Val Lys Asp His Glu Asp Ser Ser Leu Gln Trp Ser Asn Pro Ala 50 55 60 Gln Gln Thr Leu Tyr Phe Gly Glu Lys Arg Ala Leu Arg Asp Asn Arg 65 70 75 80 Ile Gln Leu Val Ser Ser Thr Pro His Glu Leu Ser Ile Ser Ile Ser 85 90 95 Asn Val Ala Leu Ala Asp Glu Gly Glu Tyr Thr Cys Ser Ile Phe Thr 100 105 110 Met Pro Val Arg Thr Ala Lys Ser Leu Val Thr Val Leu Gly Ile Pro 115 120 125 Gln Lys Pro Ile Ile Thr Gly Tyr Lys Ser Ser Leu Arg Glu Lys Glu 130 135 140 Thr Ala Thr Leu Asn Cys Gln Ser Ser Gly Ser Lys Pro Ala Ala Gln 145 150 155 160 Leu Thr Trp Arg Lys Gly Asp Gln Glu Leu His Gly Asp Gln Thr Arg 165 170 175 Ile Gln Glu Asp Pro Asn Gly Lys Thr Phe Thr Val Ser Ser Ser Val 180 185 190 Ser Phe Gln Val Thr Arg Glu Asp Asp Gly Ala Asn Ile Val Cys Ser 195 200 205 Val Asn His Glu Ser Leu Lys Gly Ala Asp Arg Ser Thr Ser Gln Arg 210 215 220 Ile Glu Val Leu Tyr Thr Pro Thr Ala Met Ile Arg Pro Glu Pro Ala 225 230 235 240 His Pro Arg Glu Gly Gln Lys Leu Leu Leu His Cys Glu Gly Arg Gly 245 250 255 Asn Pro Val Pro Gln Gln Tyr Val Trp Val Lys Glu Gly Ser Glu Pro 260 265 270 Pro Leu Lys Met Thr Gln Glu Ser Ala Leu Ile Phe Pro Phe Leu Asn 275 280 285 Lys Ser Asp Ser Gly Thr Tyr Gly Cys Thr Ala Thr Ser Asn Met Gly 290 295 300 Ser Tyr Thr Ala Tyr Phe Thr Leu Asn Val Asn Asp Pro Ser Pro Val 305 310 315 320 Pro Ser Ser Ser Ser Thr Tyr His Gly Thr Leu Glu Val Leu Phe Gln 325 330 335 Gly Pro Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys Gly Leu Val 340 345 350 Gln Pro Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu 355 360 365 His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe 370 375 380 Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr Ile Asp Gly Asp 385 390 395 400 Val Lys Leu Thr Gln Ser Met Ala Ile Ile Arg Tyr Ile Ala Asp Lys 405 410 415 His Asn Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu Ile Ser Met 420 425 430 Leu Glu Gly Ala Val Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala 435 440 445 Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu 450 455 460 Pro Glu Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr 465 470 475 480 Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala 485 490 495 Leu Asp Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro 500 505 510 Lys Leu Val Cys Phe Lys Lys Arg Ile Glu Ala Ile Pro Gln Ile Asp 515 520 525 Lys Tyr Leu Lys Ser Ser Lys Tyr Ile Ala Trp Pro Leu Gln Gly Trp 530 535 540 Gln Ala Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp 545 550 555 <210> 67 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L8_01 excluding signal sequence <400> 67 caggtgcaac ttgttcagag cggaggtggt ctcgtccaac ctggcggcag cctcagactc 60 tcttgtgctg cttcaggacg aactttcagt aattacgcac gaggatggtt cagacaggca 120 cccgggaagg ggcgcgagtt tgtggcagca atagattatt ctggtggaag caccaactac 180 gctgattctg ccaagggcag gtttaccata agtagagaca actccaagaa tactctttat 240 ttgcaaatga actcactgag agcagaggat acagccgtgt attactgcgc tgcccctgct 300 tcacgtcgtc catcttggga tgctgatgga tatgattact ggggtcaagg tactctggta 360 actgttagtt cc 372 <210> 68 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L8_01 excluding signal sequence <400> 68 Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Asn Tyr 20 25 30 Ala Arg Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Phe Val 35 40 45 Ala Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala 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 Ala Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 69 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L8_02 excluding signal sequence <400> 69 caagtccaac ttgtccaaag tggcggtggg ttggtccagc ccggcggttc tttgaggttg 60 tcatgcgccg cctccggcag gaccttctca aattacgccc gtggttggtt ccgtcaggca 120 cctgggaaag aacggggagtt cgtagctgca atagattaca gcggtgggtc aactaattac 180 gctgattctg ccaaaggaag attcaccatc tcaagagaca attctaagaa cacactttac 240 cttcagatga actctctgag agctgaagac accgctgtgt attactgtgc tgcacccgca 300 tcacggcgac cctcatggga tgctgatggg tacgactatt gggggcaagg tacacttgtt 360 actgtatcta gt 372 <210> 70 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L8_02 excluding signal sequence <400> 70 Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Asn Tyr 20 25 30 Ala Arg Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45 Ala Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala 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 Ala Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 71 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L8_03 excluding signal sequence <400> 71 caagttcaac ttgtagagtc tggaggcggt ctggttcaac ctggtgggtc cctccgcctt 60 tcctgcgctg ctagcgggag aacctttagt aattatgcac gtggctggtt taggcaggca 120 ccagggaaag ggcgtgagtt cgtcgcagca atagattata gcggcggatc taccaactac 180 gccgattcag ctaagggacg atttacaatt tcacgagaca attccaagaa taccgtttac 240 ctgcaaatga atagtctccg ggccgaagat accgctgtgt attattgtgc agcccctgct 300 tcccgccgtc ccagttggga cgcagacggg tatgactatt ggggccaggg aactttggta 360 accgtttcat ca 372 <210> 72 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L8_03 excluding signal sequence <400> 72 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 Arg Thr Phe Ser Asn Tyr 20 25 30 Ala Arg Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Phe Val 35 40 45 Ala Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val 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 Ala Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 73 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L8_04 excluding signal sequence <400> 73 caggttcagt tggttgagag cggtggtggt ctggtacagc ccggcggtag cttgcgactt 60 tcctgtgcag ccagtggtcg gacattttct aactatgccc gaggctggtt tcgccaggcc 120 cccggaaagg aacgtgagtt cgttgcagct atagattact ccggaggatc aaccaattat 180 gccgattctg caaaaggacg ctttaccatc tcccgtgaca atagtaaaaa taccgtgtac 240 ttgcaaatga acagcttgag ggcagaggat accgctgtttattactgcgc cgctcccgct 300 agtcgcaggc catcctggga cgcagatggg tatgattact ggggccaagg caccctcgta 360 actgtttcct cc 372 <210> 74 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L8_04 excluding signal sequence <400> 74 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 Arg Thr Phe Ser Asn Tyr 20 25 30 Ala Arg Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45 Ala Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val 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 Ala Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 75 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11_01 excluding signal sequence <400> 75 gaggtccaac ttgtagagtc tggaggggga ttgattcaac ccggcgggag tcttagactt 60 agctgtgccg catcaggggag cacagtgtca ttcaatgcta tggggtggta tagacaagca 120 cctgggaaag gtcttggtct ggtagccgtc atcacttctg gtgggtacac caattatgcc 180 gacagcgtca aaggccgttt taccattagt cgtgacaaca gcaagaatac cctctttctg 240 caaatgaaca gccttagagc tgaagacaca gccgtatact attgtaatgc cgagggggta 300 tattcagact atgttattat gaattattgg ggtcaaggca ctctcgttac cgtaagttca 360 <210> 76 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_01 excluding signal sequence <400> 76 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Val Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gly Leu Gly Leu Val 35 40 45 Ala Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 77 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11_02 excluding signal sequence <400> 77 gaggtacagt tggtggagag tggtggcgga ttgatccaac caggggggag cctgcgactc 60 tcctgtgctg ccagcggatc tacagtctct tttaatgcca tgggttggta tcgacaggct 120 ccaggtaaag gacgggtttt ggtcgcagta attactagcg gaggatacac aaactacgca 180 gactctgtca aggggcggtt tacaatatct cgggataact ccaagaacac cgtctatctt 240 caaatgaata gtttgcgggc cgaagatact gctgtctatt actgcaatgc tgaaggtgtg 300 tattccgatt atgttataat gaactattgg ggccagggca ccctggtcac agttagcagc 360 <210> 78 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_02 excluding signal sequence <400> 78 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Val Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gly Arg Val Leu Val 35 40 45 Ala Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 79 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11_03 excluding signal sequence <400> 79 gaagttcagt tggtagaatc cgggggaggt ttgattcaac ccggtggggag ccttagattg 60 agctgtgcag ccagcggctc aaccgtatct tttaacgcta tgggttggta tcggcaagcc 120 ccaggcaaac aaaggggttt ggtcagcgtc attaccagtg gtggttacac aaactacgca 180 gattcagtta agggccgctt cacaatctcc cgcgacaatt ccaaaaacac tgtgtatttg 240 caaatgaata gcttgagggc tgaagacaca gcagtatatt actgcaatgc tgagggtgta 300 tattctgact acgtaatcat gaactactgg ggacaaggca ctctggtgac cgtgagtagt 360 <210> 80 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_03 excluding signal sequence <400> 80 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Val Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Gly Leu Val 35 40 45 Ser Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 81 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11_04 excluding signal sequence <400> 81 gaagtccaac tggtagagag cggtgggggc cttattcagg caggaggctc tcttcgtctt 60 tcttgcgccg ccagcggcag tatcgttagc tttaatgcca tgggttggta tcgacaggcc 120 cctgggaaac aaagggggtt ggtcgcagta ataaccagtg gagggtacac caattatgca 180 gattctgtca agggaagatt caccatatca agggacaaca gtaagaacac attgtttctt 240 caaatgaata gtttgcgtgc agaagacaca gcagtgtact attgtaacgc tgagggcgtg 300 tactccgact atgttattat gaattactgg ggtcaaggta cactggtcac agttagcagc 360 <210> 82 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_04 excluding signal sequence <400> 82 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Val Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Gly Leu Val 35 40 45 Ala Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 83 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11_05 excluding signal sequence <400> 83 gaggttcagc tcgtagaaag tggggggggc ctgatacagc caggcgggag ccttagattg 60 agttgtgccg catccgggtc catattttca tttaacgcca tgggttggta cagacaagca 120 ccaggcaaag ggcgcgtatt ggtagctgtt atcaccagtg gtgggtacac aaactacgcc 180 gatagtgtta aagggcgatt tacaatatcc agagacaatt ccaaaaatac cgtttacctc 240 caaatgaata gccttagagc tgaggacact gctgtatact attgcaacgc tgagggcgta 300 tactccgatt acgtgataat gaactactgg ggccaaggca ctctggtcac cgtgtcatcc 360 <210> 84 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_05 excluding signal sequence <400> 84 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gly Arg Val Leu Val 35 40 45 Ala Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 85 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VHH of iCADM3_3R1-L11_06 excluding signal sequence <400> 85 caggttcaac tcgttgaatc tggtggaggg ttggtccagg caggggggcag tttgagactg 60 agctgcgccg catccggctc tattttctca tttaacgcca tggggtggta tcgacaggca 120 ccaggtaagc aacgcggtct cgttgcagtg ataaccagtg ggggctatac aaactatgct 180 gatagtgtta aaggcaggtt caccatcagt cgggacaaca gcaagaacac cgtcttcttg 240 caaatgaatt ctcttagagc tgaagatact gctgtatatt attgcaacgc cgagggtgtg 300 tattccgatt acgtgataat gaactactgg gggcagggga cacttgtgac cgttagttca 360 <210> 86 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_06 excluding signal sequence <400> 86 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Phe Asn 20 25 30 Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Gly Leu Val 35 40 45 Ala Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Phe Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95 Ala Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 87 <211> 375 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM 3219 excluding signal sequence <400> 87 caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc 60 tcctgcaagg cctctggata cagcttcacc ggctactata tacactgggt gcgacaggcc 120 cctggacaag gacttgagtg gatgggacgg atcaacccta acagtggtgg cacaacttat 180 gcaccgaagt ttcagggcag gttcaccatg accagagaca cgtccacgac cacagtgtac 240 ttggaactga gcggcctgag atctgaggac acggccgtgt attactgtgc gagagttctg 300 gaacgacagg gcaggccctt cgaggctgat gcttttgata tctggggcca agggacaatg 360 gtcaccgtct cttca 375 <210> 88 <211> 125 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM 3219 excluding signal sequence <400> 88 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 Gly Tyr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asn Pro Asn Ser Gly Gly Thr Thr Tyr Ala Pro Lys Phe 50 55 60 Gln Gly Arg Phe Thr Met Thr Arg Asp Thr Ser Thr Thr Thr Val Tyr 65 70 75 80 Leu Glu Leu Ser Gly Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Leu Glu Arg Gln Gly Arg Pro Phe Glu Ala Asp Ala Phe 100 105 110 Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 <210> 89 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3219 <400> 89 Gly Tyr Tyr Ile His 1 5 <210> 90 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3219 <400> 90 Arg Ile Asn Pro Asn Ser Gly Gly Thr Thr Tyr Ala Pro Lys Phe Gln 1 5 10 15 Gly <210> 91 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3219 <400> 91 Val Leu Glu Arg Gln Gly Arg Pro Phe Glu Ala Asp Ala Phe Asp Ile 1 5 10 15 <210> 92 <211> 327 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VL of CADM3219 excluding signal sequence <400> 92 gaaatagtgt 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 gctcacctcc gtggacgttc 300 ggccaaggga ccaaggtgga aataaaa 327 <210> 93 <211> 109 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VL of CADM3219 excluding the signal sequence <400> 93 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 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 Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 94 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR1 of CADM3219 <400> 94 Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala 1 5 10 <210> 95 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR2 of CADM3219 <400> 95 Gly Ala Ser Ser Arg Ala Thr 1 5 <210> 96 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR3 of CADM3219 <400> 96 Gln Gln Tyr Gly Ser Ser Pro Pro Trp Thr 1 5 10 <210> 97 <211> 378 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3301 excluding signal sequence <400> 97 cagatgcagc tggtgcaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cttctggagg caccttcaac aactatgcta tcagctgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggaggg aatattcctc tttctggaac accaaagtac 180 gcacagaagt ttcagggcag aatcacgatg accgcggaca aatccacgag cacagagtac 240 atggaactga gcagcctgac atctgaggac acggccgtat actactgtgc gagagatacc 300 ccgagtggct acaattcccc ctactactat aaaggaatgg acgtctgggg ccaagggacc 360 atggtcaccg tctcttca 378 <210> 98 <211> 126 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3301 excluding the signal sequence <400> 98 Gln Met 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 Asn Asn 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 Asn Ile Pro Leu Ser Gly Thr Pro Lys Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Ile Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Glu Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Thr Pro Ser Gly Tyr Asn Ser Pro Tyr Tyr Tyr Lys Gly 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 <210> 99 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3301 <400> 99 Asn Tyr Ala Ile Ser 1 5 <210> 100 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3301 <400> 100 Gly Asn Ile Pro Leu Ser Gly Thr Pro Lys Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 101 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3301 <400> 101 Asp Thr Pro Ser Gly Tyr Asn Ser Pro Tyr Tyr Tyr Lys Gly Met Asp 1 5 10 15 Val <210> 102 <211> 378 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3309 excluding signal sequence <400> 102 gaggtgcagc tggtgcagtc tggggctgag atgaagaagc ctgggtcctc ggtgaagctc 60 tcctgcaaat tttctggagg cgacttcagg agttatccta tcagctgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggcggc atcatcccga tttttagtcg agtaaactat 180 gcacagagat tcctgggcag aatcacgatt accgcggacg aatccacgag cacagcctac 240 atggaattga gaagcctgac gtctgacgac acggccgtct attactgtgc gacagatacc 300 ccgagtggct acaactcccc ctactactat aaaggaatgg acgtctgggg ccaggggacc 360 ctggtcaccg tctcctca 378 <210> 103 <211> 126 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3309 excluding signal sequence <400> 103 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Met Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Phe Ser Gly Gly Asp Phe Arg Ser Tyr 20 25 30 Pro Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Ser Arg Val Asn Tyr Ala Gln Arg Phe 50 55 60 Leu Gly Arg Ile Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Asp Thr Pro Ser Gly Tyr Asn Ser Pro Tyr Tyr Tyr Lys Gly 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 104 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3309 <400> 104 Ser Tyr Pro Ile Ser 1 5 <210> 105 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3309 <400> 105 Gly Ile Ile Pro Ile Phe Ser Arg Val Asn Tyr Ala Gln Arg Phe Leu 1 5 10 15 Gly <210> 106 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3309 <400> 106 Asp Thr Pro Ser Gly Tyr Asn Ser Pro Tyr Tyr Tyr Lys Gly Met Asp 1 5 10 15 Val <210> 107 <211> 351 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3312 excluding signal sequence <400> 107 cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60 acctgcagtg tctctggtgg ctccatcaga ggacactatt ggagttggat ccggcagccc 120 ccagggaagg gactggagtg gatgggttac atcaaccaca ttggggagcgc cgcctacaac 180 ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240 aagctgagct ctgtgaccgc cgcagacacg gccgtgtatt actgtgcgag aatggggcca 300 tggtgggagc ttgactactg gggccaggga accctggtca ccgtctcctc a 351 <210> 108 <211> 117 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3312 excluding the signal sequence <400> 108 Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ser Val Ser Gly Gly Ser Ile Arg Gly His 20 25 30 Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn His Ile Gly Ser Ala Ala Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Met Gly Pro Trp Trp Glu Leu Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 109 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3312 <400> 109 Gly His Tyr Trp Ser 1 5 <210> 110 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3312 <400> 110 Tyr Ile Asn His Ile Gly Ser Ala Ala Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 111 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3312 <400> 111 Met Gly Pro Trp Trp Glu Leu Asp Tyr 1 5 <210> 112 <211> 354 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3314 excluding signal sequence <400> 112 caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctggggggtc cctgagactt 60 tcctgtgcag cgtctggatt cagtttcaat aatcatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtgacattt atccggtttg atggaagtag taaatactat 180 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa caccgtgtat 240 ctggaaatga acagcctgag agcagaggac acgggtgtgt attactgtgt gaatacgcca 300 aggggttggt ccttcgatat ctggggccgt ggcaccctgg tcactgtctc ctca 354 <210> 113 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3314 excluding the signal sequence <400> 113 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Asn Asn His 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Thr Phe Ile Arg Phe Asp Gly Ser Ser Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Glu Met Asn Ser Leu Arg Ala Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95 Val Asn Thr Pro Arg Gly Trp Ser Phe Asp Ile Trp Gly Arg Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 114 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3314 <400> 114 Asn His Gly Met His 1 5 <210> 115 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3314 <400> 115 Phe Ile Arg Phe Asp Gly Ser Ser Lys Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 116 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3314 <400> 116 Thr Pro Arg Gly Trp Ser Phe Asp Ile 1 5 <210> 117 <211> 384 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3316 excluding signal sequence <400> 117 gaggtgcagc tggtggagac tgggggagcc ttggtacagc ctggggggtc cctaagactc 60 tcctgtgcag cctctggatt cacctttagc agctattcca tgaactgggt ccgccaggct 120 ccagggaagg ggctggagtg gctctcaggt attagtggtg gtgcttttag cacacactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gagagtaggt 300 cggttgagtg ggagctacaa cagatactac tactactacg gtatggacgt ctggggccaa 360 gggaccctgg tcaccgtctc ctca 384 <210> 118 <211> 128 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3316 excluding the signal sequence <400> 118 Glu Val Gln Leu Val Glu Thr Gly Gly Ala 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 Ser Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Ser Gly Ile Ser Gly Gly Ala Phe Ser Thr His 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 Val Gly Arg Leu Ser Gly Ser Tyr Asn Arg Tyr Tyr Tyr Tyr 100 105 110 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 119 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3316 <400> 119 Ser Tyr Ser Met Asn 1 5 <210> 120 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3316 <400> 120 Gly Ile Ser Gly Gly Ala Phe Ser Thr His Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 121 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3316 <400> 121 Val Gly Arg Leu Ser Gly Ser Tyr Asn Arg Tyr Tyr Tyr Tyr Tyr Gly 1 5 10 15 Met Asp Val <210> 122 <211> 357 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of CADM3349 VH excluding signal sequence <400> 122 caggtgcagc tacagcagtg gggcggaggt ctgttgacgc cttcggagac cctgtccctc 60 agctgcgatg tctctggtgg ggccttcact aattaccact ggacctggat ccgccagccc 120 ccaggaaagg gactggaatg gattggagaa atctttcata ctgggaccac caactacaac 180 ccgtccctcc agggtcgagt cgccatgtct attgacacca ccaagcggca gttcttcctg 240 aggctgacgt ctctgaccgc cgcggacacg gctgtatatt actgtgcgag agttggtaaa 300 tatggctggt acgtaggtga cttttggggc cagggaacca cggtcaccgt ctcctca 357 <210> 123 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3349 excluding signal sequence <400> 123 Gln Val Gln Leu Gln Gln Trp Gly Gly Gly Leu Leu Thr Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Ser Cys Asp Val Ser Gly Gly Ala Phe Thr Asn Tyr 20 25 30 His Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Phe His Thr Gly Thr Thr Asn Tyr Asn Pro Ser Leu Gln 50 55 60 Gly Arg Val Ala Met Ser Ile Asp Thr Thr Lys Arg Gln Phe Phe Leu 65 70 75 80 Arg Leu Thr Ser Leu Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Val Gly Lys Tyr Gly Trp Tyr Val Gly Asp Phe Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 124 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3349 <400> 124 Asn Tyr His Trp Thr 1 5 <210> 125 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3349 <400> 125 Glu Ile Phe His Thr Gly Thr Thr Asn Tyr Asn Pro Ser Leu Gln Gly 1 5 10 15 <210> 126 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3349 <400> 126 Val Gly Lys Tyr Gly Trp Tyr Val Gly Asp Phe 1 5 10 <210> 127 <211> 372 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of CADM3351 VH excluding signal sequence <400> 127 gaagtgcagc tgttgcagtc tgggggaggc ttggtccagc ctggagggtc cctgagactc 60 tcctgtgcag cctccggact catcttcagt gaccactaca tggactgggt ccgccaggct 120 ccagggaagg gactggagtg ggtcggctct attagaaata aacgtaacgg tggctccaca 180 gaatacgccg cctctgtgaa aggcagattc agcatctcaa gagatgattc aaagaattca 240 ctgtatctgc aaatgaacag cctgaaaacc gaggacacgg ccatgtattt ctgtgccaca 300 acgcgtactg gttatcaagg cttctacggc atggacgtct ggggccaagg gaccacggtc 360 accgtctcct ca 372 <210> 128 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3351 excluding the signal sequence <400> 128 Glu Val Gln Leu Leu Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Ile Phe Ser Asp His 20 25 30 Tyr Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Ser Ile Arg Asn Lys Arg Asn Gly Gly Ser Thr Glu Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Ser Ile Ser Arg Asp Asp Ser Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Met Tyr 85 90 95 Phe Cys Ala Thr Thr Arg Thr Gly Tyr Gln Gly Phe Tyr Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 129 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3351 <400> 129 Asp His Tyr Met Asp 1 5 <210> 130 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3351 <400> 130 Ser Ile Arg Asn Lys Arg Asn Gly Gly Ser Thr Glu Tyr Ala Ala Ser 1 5 10 15 Val Lys Gly <210> 131 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3351 <400> 131 Thr Arg Thr Gly Tyr Gln Gly Phe Tyr Gly Met Asp Val 1 5 10 <210> 132 <211> 321 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VL of CADM3301, CADM3309, CADM3312, CADM3314, CADM3316, CADM3349 and CADM3351 excluding signal sequence <400> 132 gaaatagtgt 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 ggcctccgac gttcggccaa 300 gggaccaagg tggaaatcaa a 321 <210> 133 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of VL of CADM3301, CADM3309, CADM3312, CADM3314, CADM3316, CADM3349 and CADM3351 excluding signal sequence <400> 133 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 Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 134 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of LCDR1 of CADM3301, CADM3309, CADM3312, CADM3314, CADM3316, CADM3349, and CADM3351 <400> 134 Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala 1 5 10 <210> 135 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of LCDR2 of CADM3301, CADM3309, CADM3312, CADM3314, CADM3316, CADM3349, and CADM3351 <400> 135 Asp Ala Ser Asn Arg Ala Thr 1 5 <210> 136 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of LCDR3 of CADM3301, CADM3309, CADM3312, CADM3314, CADM3316, CADM3349 and CADM3351 <400> 136 Gln Gln Arg Ser Asn Trp Pro Pro Thr 1 5 <210> 137 <211> 363 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3402 excluding signal sequence <400> 137 gaggtgcagc tggtggagtc tgggggaggc ttggtccagc cgggggggtc cctgagactc 60 tcctgtgcaa cctctggatt caggttcagt atgtatggca tgcactgggt ccgccagtct 120 ccagggaagg ggctggagtg ggtctcagtt atttatagcg gtggaaacac agactacgca 180 gactccgtga agggccgatt cacaatctcc agagacaatt ccaagaacac ggtgtatctt 240 caaatgaaca gcctgagagc cgaggacacg gccgtgtatt actgtgcgag tcgtcgagta 300 gttccaggtg ttatagacta ctttgactcc tggggccagg gaaccctggt caccgtctcc 360 tca 363 <210> 138 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3402 excluding signal sequence <400> 138 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 Thr Ser Gly Phe Arg Phe Ser Met Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Ser Arg Arg Val Val Pro Gly Val Ile Asp Tyr Phe Asp Ser Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 139 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3402 <400> 139 Met Tyr Gly Met His 1 5 <210> 140 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3402 <400> 140 Val Ile Tyr Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 141 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3402 <400> 141 Arg Arg Val Val Pro Gly Val Ile Asp Tyr Phe Asp Ser 1 5 10 <210> 142 <211> 363 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of CADM3404 VH excluding signal sequence <400> 142 gaggtgcagc tggtggagac cggggggggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggatt cgccttcagt aactatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtctcagtt atttatagcg gtggaaacac agactacgca 180 gactccgtga agggccgatt cacaatctcc agagacaatt ccaagaacac ggtgtatctt 240 caaatgaaca gcctgagagc cgaggacacg gccgtgtatt actgtgcgag tcgtcgagta 300 gttccaggtg ttatagacta ctttgactcc tggggccagg gaaccctggt cactgtctcc 360 tca 363 <210> 143 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3404 excluding signal sequence <400> 143 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Ser Arg Arg Val Val Pro Gly Val Ile Asp Tyr Phe Asp Ser Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 144 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3404 <400> 144 Asn Tyr Gly Met His 1 5 <210> 145 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3404 <400> 145 Val Ile Tyr Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 146 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3404 <400> 146 Arg Arg Val Val Pro Gly Val Ile Asp Tyr Phe Asp Ser 1 5 10 <210> 147 <211> 390 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3432 excluding signal sequence <400> 147 caggtacagc tgcagcagtc aggtccagga ctggtgaagg cctcgcagac cctctcactc 60 acctgtgcca tctccgggga cagtgtctct agcaggagtg ctgcttggga ctggataagg 120 cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc cacgtggtat 180 aatgactatg catcatctgt gagaagtcga ataagcatca accccgacac atccaagaac 240 cagttctccc tgcagctgaa ctctgtgact cccgaggaca cggctgtata ttattgtgtg 300 agagcaaata ggaagcttcc agcacctgga cagcactttt attatggtat ggacgtctgg 360 ggccaaggga ccacggtcac cgtctcctca 390 <210> 148 <211> 130 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3432 excluding signal sequence <400> 148 Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Ala Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Arg 20 25 30 Ser Ala Ala Trp Asp Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Thr Trp Tyr Asn Asp Tyr Ala 50 55 60 Ser Ser Val Arg Ser Arg Ile Ser Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Val Arg Ala Asn Arg Lys Leu Pro Ala Pro Gly Gln His 100 105 110 Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val 115 120 125 Ser Ser 130 <210> 149 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3432 <400> 149 Ser Arg Ser Ala Ala Trp Asp 1 5 <210> 150 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3432 <400> 150 Arg Thr Tyr Tyr Arg Ser Thr Trp Tyr Asn Asp Tyr Ala Ser Ser Val 1 5 10 15 Arg Ser <210> 151 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3432 <400> 151 Ala Asn Arg Lys Leu Pro Ala Pro Gly Gln His Phe Tyr Tyr Gly Met 1 5 10 15 Asp Val <210> 152 <211> 390 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3448 excluding signal sequence <400> 152 caggtacagc tgcagcagtc aggtccagga ctggtgaagc ccgcgcagac cctctcactc 60 acctgtgcca tctccggaga cagtgtctcc agcaacagtg ttgcttggaa ctgggtcagg 120 cagtccccat cgagaggcct tgagtggctg ggaaggacat attacaggtc ccagtggtat 180 aacgattatg caggatctgt gagaagtcga ataaccatca gcgcagacac atctaagaac 240 cagttctccc tgcaactgaa ctctgtgact cccgaggaca cggctcttta ttattgtgtg 300 agagcaaata ggaagcttcc agcacctgga cagcactttt attatggtat ggacgtctgg 360 ggccaaggga ccacggtcac cgtctcctca 390 <210> 153 <211> 130 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3448 excluding signal sequence <400> 153 Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ala Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30 Ser Val Ala Trp Asn Trp Val Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Gln Trp Tyr Asn Asp Tyr Ala 50 55 60 Gly Ser Val Arg Ser Arg Ile Thr Ile Ser Ala Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Leu 85 90 95 Tyr Tyr Cys Val Arg Ala Asn Arg Lys Leu Pro Ala Pro Gly Gln His 100 105 110 Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val 115 120 125 Ser Ser 130 <210> 154 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3448 <400> 154 Ser Asn Ser Val Ala Trp Asn 1 5 <210> 155 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3448 <400> 155 Arg Thr Tyr Tyr Arg Ser Gln Trp Tyr Asn Asp Tyr Ala Gly Ser Val 1 5 10 15 Arg Ser <210> 156 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3448 <400> 156 Ala Asn Arg Lys Leu Pro Ala Pro Gly Gln His Phe Tyr Tyr Gly Met 1 5 10 15 Asp Val <210> 157 <211> 363 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VH of CADM3458 excluding signal sequence <400> 157 gaggtgcagc tggtggagtc cgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggatt caccttcagt agatatggca tacactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt atttatagcg gtggaaacac agactacgca 180 gactccgtga agggccgatt cacaatctcc agagacaatt ccaagaacac ggtgtatctt 240 caaatgaaca gcctgagagc cgaggacacg gccgtgtatt actgtgcgag tcgtcgagta 300 gttccaggtg ttatagacta ctttgactcc tggggccagg gaaccctggt caccgtctcc 360 tca 363 <210> 158 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3458 excluding signal sequence <400> 158 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Tyr Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Ser Arg Arg Val Val Pro Gly Val Ile Asp Tyr Phe Asp Ser Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 159 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3458 <400> 159 Arg Tyr Gly Ile His 1 5 <210> 160 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3458 <400> 160 Val Ile Tyr Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 161 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3458 <400> 161 Arg Arg Val Val Pro Gly Val Ile Asp Tyr Phe Asp Ser 1 5 10 <210> 162 <211> 321 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VL of CADM3402, CADM3404, CADM3432, CADM3448 and CADM3458 excluding signal sequence <400> 162 gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60 atcacttgcc gggcaagtca gagcattagc agctatttaa attggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240 gaagattttg caacttacta ctgtcaacag agttacagta cccctcgaac gttcggccaa 300 gggaccaagg tggaaatcaa a 321 <210> 163 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: amino acid sequences of VL of CADM3402, CADM3404, CADM3432, CADM3448 and CADM3458 excluding signal sequence <400> 163 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 Ser Ile Ser Ser Tyr 20 25 30 Leu Asn 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 Ser Tyr Ser Thr Pro Arg 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 164 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of LCDR1 of CADM3402, CADM3404, CADM3432, CADM3448, and CADM3458 <400> 164 Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu Asn 1 5 10 <210> 165 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of LCDR2 of CADM3402, CADM3404, CADM3432, CADM3448, and CADM3458 <400> 165 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 166 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequences: Amino acid sequences of LCDR3 of CADM3402, CADM3404, CADM3432, CADM3448, and CADM3458 <400> 166 Gln Gln Ser Tyr Ser Thr Pro Arg Thr 1 5 <210> 167 <211> 357 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of CADM3501 VH excluding signal sequence <400> 167 gaggtgcagc tggtggagtc tgggggaggc ttagttcagc ctgggggggc cctgagactc 60 tcctgttcag cctccggatt caccttcagt gggtactgga tgcactgggt ccgccaagct 120 ccagggaagg ggctggagtg ggtgtcacaa attagtagta gtggtactat catagactcc 180 gcagactttg tgaagggccg attcgccgtc tccagggaca acgccaagga cttattgtat 240 ctgcaaatga acagcctgag agccgatgac acggccgtct attactgtgc gagggggcca 300 ctggcgaaga atggttttga catttggggc caagggacaa tggtcaccgt ctcttca 357 <210> 168 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VH of CADM3501 excluding the signal sequence <400> 168 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ala Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gln Ile Ser Ser Ser Gly Thr Ile Ile Asp Ser Ala Asp Phe Val 50 55 60 Lys Gly Arg Phe Ala Val Ser Arg Asp Asn Ala Lys Asp Leu Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Pro Leu Ala Lys Asn Gly Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 169 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR1 of CADM3501 <400> 169 Gly Tyr Trp Met His 1 5 <210> 170 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR2 of CADM3501 <400> 170 Gln Ile Ser Ser Ser Gly Thr Ile Ile Asp Ser Ala Asp Phe Val Lys 1 5 10 15 Gly <210> 171 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of HCDR3 of CADM3501 <400> 171 Gly Pro Leu Ala Lys Asn Gly Phe Asp Ile 1 5 10 <210> 172 <211> 339 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: base sequence of VL of CADM3501 excluding signal sequence <400> 172 gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 60 atcaactgca agtccagcca gagtgtttta tacagctcca acaataagaa ctacttagct 120 tggtaccagc agaaaccagg acagcctcct aagctgctca tttactgggc atctacccgg 180 gaatccgggg tccctgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 240 atcagcagcc tgcaggctga agatgtggca gtttattact gtcagcaata ttatagtact 300 ccgtacacttttggccaggg gaccaagctg gagatcaaa 339 <210> 173 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VL of CADM3501 excluding the signal sequence <400> 173 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30 Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 174 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR1 of CADM3501 <400> 174 Lys Ser Ser Gln Ser Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu 1 5 10 15 Ala <210> 175 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR2 of CADM3501 <400> 175 Trp Ala Ser Thr Arg Glu Ser 1 5 <210> 176 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Amino acid sequence of LCDR3 of CADM3501 <400> 176 Gln Gln Tyr Tyr Ser Thr Pro Tyr Thr 1 5 <210> 177 <211> 124 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L8_00 excluding signal sequence <400> 177 Gln Val Gln Leu Val Gln 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 Asn Tyr 20 25 30 Ala Arg Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Asp Tyr Ser Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala 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 Pro Ala Ser Arg Arg Pro Ser Trp Asp Ala Asp Gly Tyr Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 178 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: amino acid sequence of VHH of iCADM3_3R1-L11_00 excluding signal sequence <400> 178 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Phe Asn 20 25 30 Ala Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Thr Ser Gly Gly Tyr Thr Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Glu Gly Val Tyr Ser Asp Tyr Val Ile Met Asn Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 BRIEF DESCRIPTION OF THE DRAWINGS

[0157] [ Figure 1 ] Figure 1 (A) and (B) show the results of measuring the concentration of each antibody in tissues. Figure 1 (A) shows the antibody concentration in serum 3 days after antibody administration. The vertical axis represents the antibody concentration (ng / mL), and the horizontal axis represents the administered antibody. Figure 1 (B) shows the antibody concentration in brain tissue 3 days after antibody administration. The vertical axis represents the antibody concentration (ng / g brain), and the horizontal axis represents the administered antibody.

[0158] [ Figure 2 ] Figure 2 (A) and (B) show the results of measuring the concentration of each antibody in tissues. Figure 2 (A) shows the antibody concentration in serum 7 days after antibody administration. The vertical axis represents the antibody concentration (ng / mL), and the horizontal axis represents the administered antibody. Figure 2 (B) shows the antibody concentration in brain tissue 7 days after antibody administration. The vertical axis represents the antibody elution amount (ng / g brain), and the horizontal axis represents the administered antibody.

[0159] [ Figure 3 ] Figure 3 (A) and (B) show the results of measuring the concentration of each antibody in tissues. Figure 3 (A) shows the antibody concentration in serum 7 days after antibody administration. 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 7 days after antibody administration. The vertical axis represents the amount of antibody eluted (ng / g brain) and the horizontal axis represents the administered antibody. The antibody concentration is expressed as a value converted from the molar concentration using the molecular weight of the monoclonal antibody (150 kDa).

[0160] [ Figure 4 ] Figure 4 (A) and (B) show the results of imaging assessment of the migration ability of each antibody in mouse brain. Figure 4 (A) shows imaging images of the brain 9 days after antibody administration. Figure 4 (B) shows the ratio of the amount of fluorescence in the brain corrected for the fluorescence intensity of the administered antibody to the anti-AVM antibody. The vertical axis represents the ratio to the anti-AVM antibody, and the horizontal axis represents the administered antibody.

[0161] [ Figure 5 ] Figure 5 The results of imaging evaluation of the migration ability of each antibody in the mouse brain are shown, and imaging images of the brain 7 days after administration of the antibody are indicated.

[0162] [ Figure 6 ] Figure 6The results of imaging evaluation of the migration ability of each antibody in the mouse brain are shown, and the ratio of the fluorescence amount in the brain to the anti-AVM antibody is indicated, corrected for the fluorescence intensity of the administered antibody. The vertical axis represents the ratio to the anti-AVM antibody, and the horizontal axis represents the administered antibody.

[0163] [ Figure 7 ] Figure 7 (A) and (B) show the results of measuring the concentration of each antibody in tissues. Figure 7 (A) shows the antibody concentration in serum 7 days after antibody administration. The vertical axis represents the antibody concentration (ng / mL), and the horizontal axis represents the administered antibody. Figure 7 (B) shows the antibody concentration in brain tissue 7 days after antibody administration. The vertical axis represents the antibody elution amount (ng / g brain), and the horizontal axis represents the administered antibody.

[0164] [ Figure 8 ] Figure 8 (A) and (B) show the results of imaging assessment of the migration ability of each antibody in mouse brain. Figure 8 (A) shows imaging images of the brain 7 days after antibody administration. Figure 8 (B) shows the ratio of the amount of fluorescence in the brain corrected for the fluorescence intensity of the administered antibody to the anti-AVM antibody. The vertical axis represents the ratio to the anti-AVM antibody, and the horizontal axis represents the administered antibody.

[0165] Description of Implementation

[0166] The present invention relates to antigen-binding molecules that bind to CADM3. More specifically, the present invention relates to antibodies or antibody fragments thereof that bind to CADM3.

[0167] The CADM3-binding molecules of the present invention can be in any molecular form, as long as they specifically bind to CADM3 and the resulting molecules are retained in the brain, and can be any molecule, such as a protein, nucleic acid, or a low-molecular-weight compound or high-molecular-weight compound obtained by organic synthesis. Specifically, the CADM3-binding molecule can be any of a recombinant protein, an antibody, an aptamer, a low-molecular-weight compound obtained by low-molecular-weight screening, etc., but antibodies and antibody fragments thereof are preferably exemplified. The CADM3-binding molecule is preferably a molecule that binds to the extracellular domain of CADM3.

[0168] CADM3 is a calcium-independent immunoglobulin-like cell adhesion molecule that exhibits cell-cell adhesion activity through calcium-independent homophilic binding. For example, the full-length human CADM3, including its signal sequence, consists of 398 amino acids and is expressed at contact sites between axon terminals, between axon terminals and axon shafts, and between axon terminals and glial processes at axon terminals in the peripheral and central nervous systems, where it plays a role in cell adhesion.

[0169] The CADM3-binding molecules of the present invention bind to CADM3 in animal species such as mice, rats, cynomolgus monkeys, and / or humans, but are not particularly limited to these species. Suitable animal species can be selected based on the intended use of the antibody. For example, when the antibodies of the present invention are used for medical purposes in humans, the antibodies are preferably antibodies that bind to at least human CADM3.

[0170] In the present invention, examples of human CADM3 include a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 52 or the amino acid sequence of NCBI Accession No. AAH33819, a polypeptide consisting of an amino acid sequence in which one or more amino acids are deleted, substituted or added in the amino acid sequence represented by SEQ ID NO: 52 or the amino acid sequence of NCBI Accession No. AAH33819 and having the function of human CADM3, a polypeptide consisting of an amino acid sequence having 60% or more, preferably 80% or more, more preferably 90% or more, and most preferably 95% or more homology to the amino acid sequence represented by SEQ ID NO: 52 or the amino acid sequence of NCBI Accession No. AAH33819 and having the function of human CADM3, and the like.

[0171] The polypeptide having an amino acid sequence in which one or more amino acids are deleted, substituted or added in the amino acid sequence represented by SEQ ID NO: 52 or the amino acid sequence represented by NCBI Accession No. AAH33819 can be produced, for example, by using a site-directed mutagenesis method [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989); Current Protocols in Molecular Biology, John Wiley & Sons (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)] can be obtained by introducing site-specific mutations into a DNA encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 52.

[0172] The number of amino acids to be deleted, substituted or added is not particularly limited, but is preferably one to several tens, such as 1 to 20, more preferably one to several, such as 1 to 5 amino acids.

[0173] The same applies to the amino acid sequence of mouse CADM3 [SEQ ID NO: 54 or NCBI Accession No. NP_444429.1], the amino acid sequence of rat CADM3 [NCBI Accession No. AAI61811.1], and the amino acid sequence of cynomolgus monkey CADM3 [SEQ ID NO: 56 or NCBI Accession No. NP_001270618.1].

[0174] In the present invention, as the gene encoding human CADM3, the nucleotide sequence represented by SEQ ID NO: 51 or the nucleotide sequence of NCBI Accession No. BC033819.1 is exemplified. The present invention also includes a gene consisting of a nucleotide sequence in which one or more nucleotides are deleted, substituted or added in the nucleotide sequence represented by SEQ ID NO: 51 or the nucleotide sequence of NCBI Accession No. BC033819.1 and comprising a DNA encoding a polypeptide having the function of CADM3; a gene consisting of a nucleotide sequence having at least 60% or higher homology, preferably 80% or higher homology, more preferably 95% or higher homology to the nucleotide sequence represented by SEQ ID NO: 51 or the nucleotide sequence of NCBI Accession No. BC033819.1 and comprising a DNA encoding a polypeptide having the function of CADM3; a gene consisting of a DNA which hybridizes with a DNA comprising the nucleotide sequence represented by SEQ ID NO: 51 or the nucleotide sequence of NCBI Accession No. BC033819.1 under stringent conditions and encodes a polypeptide having the function of CADM3, etc.

[0175] The DNA that hybridizes under stringent conditions refers to hybridizable DNA obtained by colony hybridization, plaque hybridization, Southern blot hybridization, DNA microarray, etc. using a DNA comprising the nucleotide sequence represented by SEQ ID NO: 51 or the nucleotide sequence of NCBI Accession No. BC033819.1 as a probe.

[0176] Specifically, there can be exemplified a DNA that can be identified by performing a hybridization method at 65°C in the presence of 0.7 to 1.0 mol / L sodium chloride using a filter or microscope slide on which DNA derived from a colony or plaque to be hybridized or a PCR product or oligoDNA having the sequence is immobilized [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989); Current Protocols in Molecular Biology, John Wiley & Sons (1987-1997); DNA Cloning 1: Core Techniques, A Practical Approach, 2nd edition, Oxford University (1995)], and then the filter or microscope slide is washed at 65°C using a 0.1 to 2 times concentration saline sodium citrate (SSC) solution (the composition of a 1 times concentration SSC solution consists of 150 mmol / L sodium chloride and 15 mmol / L sodium citrate).

[0177] As the hybridizable DNA, there can be exemplified a DNA having at least 60% or higher homology to the nucleotide sequence represented by SEQ ID NO: 51 or the nucleotide sequence of NCBI Accession No. BC033819.1, preferably a DNA having 80% or higher homology, more preferably a DNA having 95% or higher homology.

[0178] The same applies to the nucleotide sequence of mouse CADM3 [SEQ ID NO: 53 or NCBI accession number NM_053199.3], the nucleotide sequence of rat CADM3 [NCBI accession number NM_001047103.1], and the nucleotide sequence of cynomolgus monkey CADM3 [SEQ ID NO: 55 or NCBI accession number NM_001283689.1].

[0179] Examples of the functions of CADM3 include participation in cell adhesion between axon terminals and other sites in the central nervous system and peripheral nervous system as described above, and the like.

[0180] Genetic polymorphisms are often observed in the nucleotide sequences of protein-encoding genes in eukaryotes. Genes that have undergone small-scale mutations in their nucleotide sequences due to such polymorphisms in the genes used in the present invention are also included in the genes encoding CADM3 in the present invention.

[0181] In the present invention, unless otherwise specified, the numerical value of homology may be a value calculated using a homology search program known to those skilled in the art, however, for nucleotide sequences, examples are values ​​calculated using the default parameters in BLAST [J. Mol. Biol., 215, 403 (1990)] and the like, and for amino acid sequences, examples are 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] and the like.

[0182] Regarding the default parameters, G (cost of opening a gap) is 5 in the case of nucleotide sequences and 11 in the case of amino acid sequences; -E (cost of extending a gap) is 2 in the case of nucleotide sequences and 1 in the case of amino acid sequences; -q (nucleotide mismatch penalty) is -3; -r (nucleotide match reward) is 1, and -e (expectation) is 10; -W (wordlength) is 11 in the case of nucleotide sequences and 3 in the case of amino acid sequences; -y [decay (X) in bits for blast extension] is 20 in the case of blastn and 7 in the case of programs other than blastn; -X (X decay value in bits for gapped alignments) is 15; and -Z (final X decay value in bits for gapped alignments) is 50 in the case of blastn and 25 in the case of programs other than blastn (http: / / www.ncbi.nlm.nih.gov / blast / html / blastcgihelp.html).

[0183] Polypeptides comprising a partial sequence of the amino acid sequence of any of the various types of CADM3 mentioned above can be produced by methods known to those skilled in the art. Specifically, the polypeptides can be produced by deleting a portion of the DNA encoding the amino acid sequence of any of the various types of CADM3 mentioned above, and culturing transformants transfected with an expression vector containing the resulting DNA. Furthermore, polypeptides having an amino acid sequence in which one or more amino acids in the amino acid sequence of any of the various types of CADM3 are deleted, substituted, or added can be obtained in the same manner as described above.

[0184] In addition, a polypeptide consisting of the amino acid sequence of any one of the various different types of CADM3 or a polypeptide having an amino acid sequence in which one or more amino acids are deleted, substituted or added in the amino acid sequence of any one of the various different types of CADM3 can also be produced by chemical synthesis methods such as the fluorenylmethoxycarbonyl (Fmoc) method or the tert-butoxycarbonyl (tBoc) method.

[0185] In the present invention, the extracellular domain of human CADM3 refers to the amino acid sequence from positions 25 to 330 in the amino acid sequence represented by SEQ ID NO: 52 or NCBI Accession No. AAH33819.

[0186] The extracellular domain of mouse CADM3 refers to the amino acid sequence from positions 23 to 328 in the amino acid sequence represented by SEQ ID NO: 54 or NCBI Accession No. NP_444429.1. The extracellular domain of rat CADM3 refers to the amino acid sequence from positions 23 to 328 in the amino acid sequence represented by NCBI Accession No. AAI61811.1.

[0187] The extracellular domain of cynomolgus monkey CADM3 refers to the amino acid sequence from position 23 to position 328 in the amino acid sequence represented by SEQ ID NO: 56 or NCBI Accession No. NP_001270618.1.

[0188] Binding of the CADM3-binding molecules of the present invention to the extracellular domain of CADM3 can be confirmed by measuring the affinity of the CADM3-binding molecules of the present invention for CADM3-expressing cells or recombinant CADM3 protein using enzyme-linked immunosorbent assay (ELISA), flow cytometry, surface plasmon resonance, etc. Furthermore, this can be confirmed by combining known immunoassay methods [Monoclonal Antibodies - Principles and Practice, 3rd Edition, Academic Press (1996); Antibodies - A Laboratory Manual, Cold Spring Harbor Laboratory (1988); Manual for monoclonal antibody experiments, Kodansha Scientific Books (1987)].

[0189] The CADM3-binding molecules of the present invention are molecules that accumulate in the brain by specifically binding to CADM3 in the brain. For example, the antibodies of the present invention are antibodies that accumulate in the brain by binding to CADM3 in the brain. Furthermore, when the antibodies are administered to an animal in the periphery, they accumulate in the brain by crossing the blood-brain barrier in the brain from the periphery, migrating to the brain, and binding to CADM3 in the brain. The antibodies of the present invention are preferably antibodies that have excellent or enhanced brain accumulation.

[0190] In the present invention, the "property of accumulating in the brain" refers to a property in which the target agent is retained in the brain when administered to a test animal. In other words, it means that the concentration of the target agent in the brain (or the amount in the brain) increases, or the target agent is present at a fixed concentration, so that it can be detected due to at least one reason selected from the following: increased migration into the brain, increased accumulation in the brain, decreased migration from the inside of the brain to the outside, decreased excretion from the inside of the brain to the outside, and decreased decomposition in the brain.

[0191] In the present invention, “having an excellent property of accumulating in the brain”, “having a high property of accumulating in the brain” or “having an enhanced property of accumulating in the brain” means that when the target substance is administered to a test animal, the concentration of the target substance in the brain (or the amount in the brain) is increased compared to the control after the same number of days from administration, or the target substance is present at a fixed concentration (amount) so that it can be detected in the brain for a long time.

[0192] This phenomenon occurs due to at least any of the following reasons: increased migration of the target substance into the brain, increased accumulation in the brain, decreased migration from the inside to the outside of the brain, decreased efflux from the inside to the outside of the brain, and decreased decomposition in the brain compared to the control.

[0193] In the present invention, “having an excellent property of accumulating in the brain”, “having a high property of accumulating in the brain” or “having an enhanced property of accumulating in the brain” include, for example, when the target subject is administered to a test animal, the concentration (amount) of the target subject in the brain is higher than that of the control 1 to 10 days after administration, preferably 2 to 10 days, 3 to 10 days, more preferably 4 to 10 days after administration, or the concentration of the target subject in the brain (or the amount in the brain) reaches its peak on the 4th day or later, preferably on the 5th day or later, 6th day or later, 7th day or later, 8th day or later, 9th day or later, more preferably 10th day or later after administration, and the like.

[0194] The antibody having an excellent property of accumulating in the brain, the antibody having a high property of accumulating in the brain, or the antibody having an enhanced property of accumulating in the brain may be any antibody as long as the antibody concentration (antibody amount) in the brain is higher than that of a control antibody or the antibody has the characteristic of being able to exist in the brain for a long time.

[0195] For example, examples include antibodies having characteristics that the ability to migrate into the brain and / or the ability to accumulate in the brain is higher than that of a control antibody, characteristics that the ability to migrate from the inside of the brain to the outside, the ability to be excreted, and / or the ability to be degraded in the brain are lower than those of a control antibody, and characteristics that the ability to migrate into the brain and / or the ability to accumulate in the brain is higher than the ability to migrate from the inside of the brain to the outside, the ability to be excreted, and / or the ability to be degraded in the brain, etc.

[0196] Therefore, examples of the antibodies or antibody fragments thereof of the present invention include antibodies or antibody fragments thereof that, when administered to an animal, have a higher antibody concentration (or antibody amount) in the brain than a control antibody after the same number of days have passed from administration, or antibodies or antibody fragments thereof that are able to exist in the brain for a long time, and the like.

[0197] The change in the antibody concentration (or antibody amount) in the brain may be any change, examples of which include a case where, after the antibody concentration in the brain once reaches a peak during the measurement period, the antibody concentration gradually decreases, a case where, after the antibody concentration in the brain has reached a peak, the antibody concentration continues to be maintained, or a case where the antibody concentration in the brain continues to increase after administration of the antibody, and the like.

[0198] Examples of the antibodies or antibody fragments thereof of the present invention include, for example, antibodies whose antibody concentration or antibody amount in the brain is higher than that of a control antibody on the 4th or 10th day after administration to rats, antibodies whose antibody concentration or antibody amount in the brain is maintained or increased during the period from the 4th day to the 10th day after administration to rats, or antibodies whose presence in the brain can be clearly confirmed even on the 10th day or later after administration to rats, etc., but are not limited thereto.

[0199] The control antibody may be any antibody as long as it is of the same type or subclass as the test antibody, but for example, an anti-avermectin (AVM) antibody can be used.

[0200] In the present invention, examples of “in the brain” include, but are not limited to, in the brain parenchyma, in the cerebral ventricles, in the cerebrospinal fluid, and the like.

[0201] By immunoelectron microscopy, staining of CADM3 was confirmed at, for example, parallel fiber terminals of granule cells, contact sites between parallel fiber terminals and parallel fiber axons, and contact sites between parallel fiber terminals and glial cell processes (NPL 26). Therefore, as one aspect of the CADM3-binding molecules of the present invention, examples include molecules that specifically bind to CADM3 in neurons and / or neural tissue, thereby having an affinity for neurons and thus accumulating in the brain. As one aspect of the antibodies of the present invention, examples include antibodies that, for example, bind to CADM3 in neurons and / or neural tissue, thereby having an affinity for neurons and thus accumulating in the brain.

[0202] In the present invention, as a method of administering the antibody to an animal, for example, intravenous administration, intracerebroventricular administration, intraperitoneal administration, subcutaneous administration, intradermal administration, intranasal administration, intrathecal administration and the like are exemplified, but are not limited thereto.

[0203] In the present invention, as a method for measuring the property of accumulation of an antibody in the brain, examples include, for example, a method of collecting brain tissue a few days after administration of the antibody to an animal, followed by homogenization and centrifugation, and then measuring the antibody concentration in the obtained supernatant and calculating the amount of the antibody per unit brain weight, a method of detecting the presence of the antibody by a known immunological method using the collected brain tissue, a method of administering a labeled antibody to an animal and detecting the presence of the antibody over time using an in vivo imaging system, and the like.

[0204] Examples of the antibodies or antibody fragments thereof of the present invention include antibodies or antibody fragments selected from the following (a) to (x). Among them, (d), (j), (o), or (t) is preferred from the viewpoint of the antibody's brain accumulation and the amount of antibody in the brain.

[0205] (a) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 23, 24, and 25, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 28, 29, and 30, respectively;

[0206] (b) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 34, 35, and 36, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 38, 39, and 40, respectively;

[0207] (c) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 3, 4, and 5, respectively;

[0208] (d) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 8, 9, and 10, respectively;

[0209] (e) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 13, 14, and 15, respectively;

[0210] (f) an antibody fragment, wherein the amino acid sequences of CDR1 to CDR3 of VHH comprise the amino acid sequences represented by SEQ ID NOs: 18, 19, and 20, respectively;

[0211] (g) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 89, 90, and 91, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 94, 95, and 96, respectively;

[0212] (h) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 99, 100, and 101, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0213] (i) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 104, 105, and 106, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0214] (j) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 109, 110, and 111, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0215] (k) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 114, 115, and 116, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0216] (1) An antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 119, 120, and 121, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0217] (m) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 124, 125, and 126, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0218] (n) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 129, 130, and 131, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 134, 135, and 136, respectively;

[0219] (o) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 139, 140, and 141, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0220] (p) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 144, 145, and 146, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0221] (q) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 149, 150, and 151, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0222] (r) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 154, 155, and 156, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0223] (s) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 159, 160, and 161, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 164, 165, and 166, respectively;

[0224] (t) an antibody, wherein the amino acid sequences of CDR1 to CDR3 of VH comprise the amino acid sequences represented by SEQ ID NOs: 169, 170, and 171, respectively, and wherein the amino acid sequences of CDR1 to CDR3 of VL comprise the amino acid sequences represented by SEQ ID NOs: 174, 175, and 176, respectively;

[0225] (u) an antibody that competes with at least one of the antibodies or antibody fragments described in (a) to (t) for binding to CADM3;

[0226] (v) an antibody that binds to an epitope comprising the epitope to which any one of the antibodies or antibody fragments described in (a) to (t) binds;

[0227] (w) an antibody that binds to the same epitope as that bound by any one of the antibodies or antibody fragments described in (a) to (t); and

[0228] (x) An antibody comprising an amino acid sequence having 85% or higher homology to the amino acid sequence of any one of the antibodies or antibody fragments described in (a) to (t).

[0229] The antibodies of the present invention include antibodies comprising an amino acid sequence of CDR1 to CDR3 of VH and CDR1 to CDR3 of VL of an antibody having 85% or more, preferably 90% or more homology to the amino acid sequence of CDR1 to CDR3 of VH and CDR1 to CDR3 of VL of any of the antibodies or antibody fragments described in (a) to (t). The 90% or more homology is more preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homology, etc.

[0230] In the present invention, as one case of the antibodies or antibody fragments described in (a) to (t), examples are CADM301 antibody, CADM3102 antibody, CADM3219 antibody, CADM3301 antibody, CADM3309 antibody, CADM3312 antibody, CADM3314 antibody, CADM3316 antibody, CADM3349 antibody, CADM3351 antibody, CADM3402 antibody, CADM3404 antibody, CADM3432 antibody, CADM3448 antibody, CADM3458 antibody and CADM3501 antibody, each of which is a human anti-CADM3 monoclonal antibody, and iCADM3-3R1-L5 antibody, iCADM3-3R1-L8 antibody, iCADM3-3R1-L10 antibody and iCADM3-3R1-L11 antibody, each of which is an alpaca anti-CADM3 monoclonal VHH antibody. Among them, from the viewpoint of the property of the antibody to accumulate in the brain and the amount of the antibody in the brain, the CADM3312 antibody, the CADM3402 antibody, the CADM3502 antibody, or the iCADM3-3R1-L8 antibody is preferred.

[0231] Additional examples thereof include human chimeric antibodies and humanized antibodies generated from any of the above-mentioned monoclonal antibodies by genetic recombination techniques, etc. Specific examples thereof also include iCADM3-3R1-L8_01 humanized antibody, iCADM3-3R1-L8_02 humanized antibody, iCADM3-3R1-L8_03 humanized antibody, iCADM3-3R1-L8_04 humanized antibody, iCADM3-3R1-L11_01 humanized antibody, iCADM3-3R1-L11_02 humanized antibody, iCADM3-3R1-L11_03 humanized antibody, iCADM3-3R1-L11_04 humanized antibody, iCADM3-3R1-L11_05 humanized antibody, iCADM3-3R1-L11_06 humanized antibody, and the like.

[0232] In the present invention, the antibody (u) refers to a second antibody that inhibits binding of any one of the antibodies or antibody fragments described in (a) to (t) to CADM3 when the first antibody is defined as the first antibody.

[0233] In the present invention, the antibody (w) refers to a second antibody that binds to a second epitope including the first epitope, when any one of the antibodies or antibody fragments described in (a) to (t) is defined as a first antibody and the epitope to which the first antibody binds is defined as a first epitope.

[0234] Furthermore, the antibody (x) of the present invention refers to a second antibody that binds to the first epitope when any one of the antibodies or antibody fragments described in (a) to (t) is defined as a first antibody and the epitope to which the first antibody binds is defined as a first epitope.

[0235] Furthermore, the antibodies or antibody fragments thereof of the present invention specifically include antibodies or antibody fragments selected from the following (1) to (31). Among them, (4), (20), (25), or (30) is preferred from the viewpoint of the property of accumulation of the antibody in the brain and the amount of the antibody in the brain.

[0236] (1) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 22, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 27;

[0237] (2) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 32, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 37;

[0238] (3) an antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 2;

[0239] (4) an antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 7;

[0240] (5) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 12;

[0241] (6) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 17;

[0242] (7) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 68;

[0243] (8) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 70;

[0244] (9) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 72;

[0245] (10) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 74;

[0246] (11) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 76;

[0247] (12) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 78;

[0248] (13) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 80;

[0249] (14) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 82;

[0250] (15) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 84;

[0251] (16) An antibody fragment, wherein the amino acid sequence of VHH comprises the amino acid sequence represented by SEQ ID NO: 86;

[0252] (17) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 88, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 93;

[0253] (18) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 98, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0254] (19) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 103, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0255] (20) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 108, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0256] (21) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 113, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0257] (22) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 118, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0258] (23) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 123, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0259] (24) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 128, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 133;

[0260] (25) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 138, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0261] (26) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 143, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0262] (27) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 148, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0263] (28) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 153, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0264] (29) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 158, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 163;

[0265] (30) An antibody, wherein the amino acid sequence of VH comprises the amino acid sequence represented by SEQ ID NO: 168, and wherein the amino acid sequence of VL comprises the amino acid sequence represented by SEQ ID NO: 173; and

[0266] (31) An antibody comprising an amino acid sequence having 85% or higher homology to the amino acid sequence of any one of the antibodies or antibody fragments described in (1) to (30).

[0267] The antibodies of the present invention include antibodies comprising VH and VL amino acid sequences of an antibody having 85% or higher, preferably 90% or higher homology to the VH and VL amino acid sequences of any of the antibodies or antibody fragments described in (1) to (30). The 90% or higher homology is more preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homology, etc.

[0268] In the present invention, as one case of the antibodies or antibody fragments described in (1) to (31), examples are CADM301 antibody, CADM3102 antibody, CADM3219 antibody, CADM3301 antibody, CADM3309 antibody, CADM3312 antibody, CADM3314 antibody, CADM3316 antibody, CADM3349 antibody, CADM3351 antibody, CADM3402 antibody, CADM3404 antibody, CADM3432 antibody, CADM3448 antibody, CADM3458 antibody and CADM3501 antibody, each of which is a human anti-CADM3 monoclonal antibody, and iCADM3-3R1-L5 antibody, iCADM3-3R1-L8 antibody, iCADM3-3R1-L10 antibody and iCADM3-3R1-L11 antibody, each of which is an alpaca anti-CADM3 monoclonal VHH antibody. Among them, from the viewpoint of the property of the antibody to accumulate in the brain and the amount of the antibody in the brain, the CADM3312 antibody, the CADM3402 antibody, the CADM3502 antibody, or the iCADM3-3R1-L8 antibody is preferred.

[0269] Additional examples thereof include human chimeric antibodies and humanized antibodies generated from any of the above-mentioned monoclonal antibodies by genetic recombination technology, etc. Specific examples thereof also include: a humanized antibody in which at least one amino acid residue at a position selected from the group consisting of position 6, 27, 37, 44, 45, 47, 49, 79, and 98 in the amino acid sequence of SEQ ID NO: 177 is substituted; a humanized antibody in which at least one amino acid residue at a position selected from the group consisting of position 1, 12, 14, 27, 28, 29, 37, 44, 45, 46, 47, 49, 78, 96, and 97 in the amino acid sequence of SEQ ID NO: 178 is substituted; a humanized antibody comprising SEQ ID NO: 179; A humanized antibody comprising at least one of the following amino acid residue substitutions in the amino acid sequence of NO: 177: the amino acid residue at position 6 is replaced with Glu, the amino acid residue at position 27 is replaced with Arg, the amino acid residue at position 37 is replaced with Phe, the amino acid residue at position 44 is replaced with Glu, the amino acid residue at position 45 is replaced with Arg, the amino acid residue at position 47 is replaced with Phe, the amino acid residue at position 49 is replaced with Ala, the amino acid residue at position 79 is replaced with Val, and the amino acid residue at position 98 is replaced with Ala; comprising SEQ ID NO: 177 a humanized antibody in which at least one of the following amino acid residue substitutions is made in the amino acid sequence of ID NO: 178: the amino acid residue at position 1 is replaced with Gln, the amino acid residue at position 12 is replaced with Val, the amino acid residue at position 14 is replaced with Ala, the amino acid residue at position 27 is replaced with Ser, the amino acid residue at position 28 is replaced with Ile, the amino acid residue at position 29 is replaced with Phe, the amino acid residue at position 37 is replaced with Tyr, the amino acid residue at position 44 is replaced with Gln, the amino acid residue at position 45 is replaced with Arg, the amino acid residue at position 46 is replaced with Gly, and the amino acid residue at position 47 is replaced with The amino acid residue at position 1 is replaced with Leu, the amino acid residue at position 49 is replaced with Ala, the amino acid residue at position 78 is replaced with Val, the amino acid residue at position 96 is replaced with Asn, and the amino acid residue at position 97 is replaced with Ala; iCADM3-3R1-L8_01 humanized antibody; iCADM3-3R1-L8_02 humanized antibody; iCADM3-3R1-L8_03 humanized antibody; iCADM3-3R1-L8_04 humanized antibody; iCADM3-3R1-L11_01 humanized antibody; iCADM3-3R1-L11_02 humanized antibody; iCADM3-3R1-L11_03 humanized antibody;iCADM3-3R1-L11_04 humanized antibody; iCADM3-3R1-L11_05 humanized antibody; iCADM3-3R1-L11_06 humanized antibody; etc.;

[0270] In the present invention, the EU index refers to the position of amino acid residues according to Sequences of Proteins of Immunological Interest, 5th edition (1991). Unless otherwise specified, the positions of amino acid residues shown below refer to the positions of amino acid residues according to the EU index.

[0271] An antibody molecule is also called an immunoglobulin (Ig), and its basic structure is a tetramer having two polypeptides called heavy chains (H chains) and two polypeptides called light chains (L chains).

[0272] Furthermore, each H chain is composed of the corresponding domains of the H chain variable domain (also called VH) and the H chain constant domain (also called CH) from the N-terminal side, and each L chain is composed of the corresponding domains of the L chain variable domain (also called VL) and the L chain constant domain (also called CL) from the N-terminal side.

[0273] As CH, α, δ, ε, γ, and μ chains are known for each subclass. The CH is further composed of the corresponding domains of the CH1 domain, hinge domain, CH2 domain, and CH3 domain from the N-terminal side.

[0274] The domain refers to the functional structural unit of each polypeptide that constitutes the antibody molecule. In addition, the CH2 domain and CH3 domain are collectively referred to as the Fc (crystallizable fragment) region or simply Fc. As CL, C λ Chain and C κ chain.

[0275] The subclasses of antibodies in which CH is an α, δ, ε, γ, and μ chain are called IgA, IgD, IgE, IgG, and IgM, respectively. Each antibody subclass sometimes has isotypes depending on the animal. In humans, there are IgA1 and IgA2 isotypes for IgA, and IgG1, IgG2, IgG3, and IgG4 isotypes for IgG.

[0276] In the present invention, the CH1 domain, hinge domain, CH2 domain, CH3 domain and Fc region can be specified according to the EU index by numbering amino acid residues from the N-terminus.

[0277] Specifically, CH1 is defined as the amino acid sequence at positions 118 to 215 according to the EU index, the hinge is defined as the amino acid sequence at positions 216 to 230 according to the EU index, CH2 is defined as the amino acid sequence at positions 231 to 340 according to the EU index, CH3 is defined as the amino acid sequence at positions 341 to 447 according to the EU index, and the Fc region is defined as the amino acid sequence at positions 231 to 447 according to the EU index.

[0278] As the antibody of the present invention, polyclonal antibodies, monoclonal antibodies and oligoclonal antibodies are all included. The polyclonal antibody refers to a group of antibody molecules secreted by antibody-producing cells of different clones. The monoclonal antibody is an antibody secreted by antibody-producing cells of a single clone, and refers to an antibody that only recognizes one epitope (also referred to as an antigenic determinant), and wherein the amino acid sequence (primary sequence) constituting the monoclonal antibody is homogeneous. The oligoclonal antibody refers to a group of antibody molecules in which a plurality of different monoclonal antibodies are mixed.

[0279] As the monoclonal antibody in the present invention, examples are antibodies produced by hybridomas or genetically recombinant antibodies produced by transformants transformed with an expression vector containing an antibody gene.

[0280] Examples of the epitope include a single amino acid sequence, a conformation constituted by an amino acid sequence, a conformation constituted by an amino acid sequence, a post-translationally modified amino acid sequence, and a conformation constituted by a post-translationally modified amino acid sequence, etc., each of which is recognized and bound by the monoclonal antibody.

[0281] As the post-translationally modified amino acid sequence, examples are O-linked glycans in which glycans are attached to Tyr and Ser having an OH substituent, N-linked glycans in which glycans are attached to Gln and Asn having an NH2 substituent, and tyrosine-sulfated amino acid sequences in which a sulfate molecule is attached to Tyr having an OH substituent.

[0282] The epitope of CADM3 to which the antibody of the present invention binds can be identified by performing an antibody binding assay using a deletion variant in which certain domains of CADM3 are deleted, a mutant in which certain domains of CADM3 are replaced with a domain derived from another protein, a partial peptide fragment of CADM3, etc. In addition, the antibody binding assay can also be performed using cells expressing the deletion variant or the mutant.

[0283] Alternatively, the epitope of CADM3 to which the antibody of the present invention binds can also be identified by adding the antibody of the present invention to a peptide fragment of CADM3 obtained by protease digestion and performing epitope mapping using known mass spectrometry.

[0284] The antibodies of the present invention also include genetically recombinant antibodies produced by genetic recombination technology, such as mouse antibodies, rat antibodies, hamster antibodies, rabbit antibodies, llama antibodies, camel antibodies, alpaca antibodies, chimeric antibodies, humanized antibodies (also known as "CDR-grafted antibodies") and human antibodies.

[0285] In the present invention, the chimeric antibody refers to an antibody in which VH and VL are derived from an animal species different from CH and CL. An antibody composed of VH and VL of an antibody from an animal other than humans (non-human animals) and CH and CL of a human antibody is called a human chimeric antibody, and an antibody composed of VH and VL of an antibody from an animal other than mice and CH and CL of a mouse antibody is called a mouse chimeric antibody. Other chimeric antibodies are also named in the same way.

[0286] As the non-human animal, any animal such as mouse, rat, hamster, rabbit, llama, camel or alpaca can be used as long as it is an animal capable of producing hybridomas or antibody phage libraries.

[0287] The hybridoma refers to a cell obtained by fusing B cells obtained by immunizing a non-human animal with an antigen with myeloma cells derived from mice or the like, and produces a monoclonal antibody having desired antigen specificity.

[0288] The antibody phage library is a library in which immunoglobulin variable region genes are cloned into phage and antigen-binding molecules are expressed on the surface of the phage. Examples of the phage used are M13 phage, but are not particularly limited.

[0289] The antigen-binding molecules displayed on phages may be in any form, but are preferably antibody fragments such as scFv, Fab, or VHH.

[0290] In the present invention, the antibody phage library can be any of an immune library, a natural library and a synthetic library.

[0291] The immune library refers to an antibody phage library constructed based on antibody genes derived from lymphocytes of animals or patients immunized with antigens. The natural library refers to an antibody phage library constructed based on antibody genes derived from lymphocytes of normal animals or healthy humans. The synthetic library refers to a library in which the CDRs of V genes or reconstructed functional V genes in genomic DNA are replaced by oligonucleotides encoding random amino acid sequences of appropriate length.

[0292] As a method for producing chimeric antibodies, a method for producing human chimeric antibodies will be described below. Other chimeric antibodies can also be produced in the same manner.

[0293] The human chimeric antibody can be produced as follows: cDNAs encoding VH and VL are obtained from a hybridoma derived from non-human animal cells producing a monoclonal antibody, each of the cDNAs is inserted into an expression vector for animal cells having DNA encoding CH and CL of a human antibody, thereby constructing a human chimeric antibody expression vector, and then the vector is introduced into animal cells and the antibody is expressed.

[0294] In addition, the human chimeric antibody can also be produced as follows: genes encoding VH and VL are cloned from an antibody phage library derived from a non-human animal, each of the genes is inserted into an expression vector for animal cells having DNA encoding CH and CL of a human antibody, thereby constructing a human chimeric antibody expression vector, and then the vector is introduced into animal cells to express the antibody.

[0295] The humanized antibody refers to an antibody in which the amino acid sequences of the CDRs of VH and VL of a non-human animal antibody are transplanted into the corresponding CDRs of VH and VL of a human antibody. The regions outside the CDRs of VH and VL are called FRs.

[0296] The humanized antibody can be produced as follows: construct a cDNA encoding the amino acid sequence of VH composed of the amino acid sequence of CDR of VH of a non-human animal antibody and the amino acid sequence of FR of VH of any human antibody, and a cDNA encoding the amino acid sequence of VL composed of the amino acid sequence of CDR of VL of a non-human animal antibody and the amino acid sequence of FR of VL of any human antibody, insert each of the cDNAs into an expression vector for animal cells having DNA encoding CH and CL of a human antibody, thereby constructing a humanized antibody expression vector, and then introduce the vector into animal cells and express the antibody.

[0297] The human antibodies originally refer to antibodies naturally existing in the human body, but also include antibodies obtained from human antibody phage libraries or transgenic animals producing human antibodies.

[0298] The human antibodies can be obtained by immunizing mice having human immunoglobulin genes with the desired antigen (Tomizuka K. et al., Proc Natl Acad Sci USA. 97, 722-7, 2000). In addition, the human antibodies can be obtained by selecting human antibodies having the desired binding activity using a phage display library obtained by amplifying antibody genes from human-derived B cells without using immunization (Winter G. et al., Annu Rev Immunol. 12: 433-55. 1994).

[0299] Furthermore, the human antibodies can be obtained by immortalizing human B cells using Epstein-Barr virus to generate cells producing human antibodies having the desired binding activity (Rosen A. et al., Nature 267, 52-54. 1977).

[0300] The human antibody phage library is a library of phages expressing antibody fragments such as Fab, scFv or VHH on the surface obtained by inserting antibody genes prepared from lymphocytes of humans (healthy humans or patients) into phage genes. Phages expressing antibody fragments with desired antigen-binding activity can be collected from the library using binding activity to a substrate with an antigen fixed thereon as an indicator. The antibody fragments can also be further converted into human antibody molecules consisting of two complete H chains and two complete L chains using genetic engineering techniques.

[0301] The transgenic animal producing human antibodies refers to an animal in which human antibody genes are incorporated into the chromosomes of the host animal. Specifically, transgenic animals producing human antibodies can be produced by introducing human antibody genes into mouse ES cells, implanting the ES cells into the early embryos of another mouse, and then allowing the embryos to develop into animals.

[0302] Production of human antibodies from the human antibody-producing transgenic animal can be performed by culturing a human antibody-producing hybridoma obtained by a general hybridoma production method using a mammal other than humans to produce and accumulate the human antibody in culture, and purifying the antibody from the culture.

[0303] The antibodies of the present invention include heavy chain antibodies consisting only of heavy chains. The heavy chain antibodies are antibodies obtained from camelids such as llamas, camels and alpacas, or genetically recombinant antibodies generated based on the antibodies.

[0304] In the present invention, the antibody fragment is an antibody fragment and refers to a fragment having antigen-binding activity. Examples include Fab, Fab', F(ab')2, scFv, diabodies, dsFv, peptides containing multiple CDRs, VHH, etc. In addition, the antibody fragment of the present invention also includes any antibody fragment, as long as the antibody fragment comprises a partial fragment of an antibody and has CADM3 binding activity, such as an antibody fragment obtained by fusing the full length or a portion of the constant region or Fc of an antibody to the antibody fragment, or an antibody fragment comprising a constant region or Fc.

[0305] The Fab is an antibody fragment obtained by treating an IgG antibody with papain (cleavage at the amino acid residue at position 224 in the H chain), has a molecular weight of about 50,000, and has antigen-binding activity, and in which about half of the H chain and the entire L chain on the N-terminal side are linked by a disulfide bond (SS bond).

[0306] The F(ab')2 is an antibody fragment obtained by treating IgG with pepsin (cleavage at the 234th amino acid residue in the H chain), has a molecular weight of about 100,000 and has antigen-binding activity, and is slightly larger than a molecule obtained by linking Fab via an SS bond in the hinge region.

[0307] The Fab' is an antibody fragment having a molecular weight of about 50,000 and antigen-binding activity, wherein the SS bond in the hinge region of the F(ab')2 is cleaved.

[0308] The scFv is a VH-P-VL or VL-P-VH polypeptide, in which one VH and one VL are linked using a suitable peptide linker (P), for example, a linker peptide obtained by linking any number of linkers consisting of 4 Gly residues and 1 Ser residue (G4S), and is an antibody fragment having antigen-binding activity.

[0309] The diabody is an antibody fragment in which scFvs having the same or different antigen-binding specificities form a dimer, and is an antibody fragment having bivalent antigen-binding activity to the same antigen or antigen-binding activity specifically to different antigens.

[0310] The dsFv is an antibody fragment obtained by linking polypeptides in which one amino acid residue in each of VH and VL is replaced by a cysteine ​​residue through an SS bond between the cysteine ​​residues, and has antigen-binding activity.

[0311] The peptide comprising CDRs is constructed to comprise at least one or more CDR regions of VH or VL and is an antibody fragment with antigen-binding activity. In a peptide comprising multiple CDRs, the CDRs may be connected directly or through a suitable peptide connector. As a peptide comprising CDRs of the present invention, an example is a peptide comprising 6 CDRs derived from an antibody of the present invention.

[0312] The peptide comprising the CDR can be produced by constructing a DNA encoding the CDRs of the VH and VL of the antibody of the present invention, inserting the DNA into an expression vector for prokaryotes or an expression vector for eukaryotes, then introducing the expression vector into the prokaryote or eukaryote and expressing the peptide. Alternatively, the peptide comprising the CDR can be produced by chemical synthesis methods such as the Fmoc method or the tBoc method.

[0313] The VHH is a variable domain of a heavy chain antibody and is also called a nanobody. The antibody fragment of the present invention includes any antibody fragment, as long as the antibody fragment includes any of the above-mentioned antibody fragments or a partial fragment thereof and has CADM3 binding activity.

[0314] In the present invention, an antibody or antibody fragment thereof having one antigen binding site is referred to as a monovalent antibody. Examples of monovalent antibody formats include the formats of antibodies or antibody fragments thereof having one antigen binding site described in WO 2014 / 054804, WO 2011 / 090754, WO 2007 / 048037, WO 2012 / 116927, and other formats.

[0315] In the present invention, an antibody or antibody fragment thereof that binds to three or more different antigens or epitopes in one molecule is referred to as a multispecific antibody. In addition, in the present invention, an antibody or antibody fragment thereof that binds to two different antigens or epitopes in one molecule is referred to as a bispecific antibody.

[0316] Examples of formats for multispecific or bispecific antibodies include those described in WO 2009 / 131239; WO 2014 / 054804; WO 01 / 077342; U.S. Patent Application Publication No. 2007 / 0071675; WO 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]; Jong et al. [http: / / dx.doi.org / 10.1371 / journal.pbio.1002344]; Kontermann et al. [mAbs 2012, vol. 4, issue 2, pp. 182-197]; Spiess et al. [Molecular Biology, 2012, vol. 4, issue 2, pp. 182-197]; and others. Immunology 67(2015)95-106]; Ridgway et al. [Proteinengineering, 1996 vol.9, no.7, pp.617-621]; WO 2009 / 080251; WO 2010 / 151792; WO 2014 / 033074, etc.

[0317] Specific examples of the bispecific antibody include the following bispecific antibodies and the like.

[0318] (1) A bispecific antibody, wherein amino acid modifications S354C / T366W are introduced into CH3 of one heavy chain (heavy chain A) of the two heavy chains of the antibody, and amino acid modifications Y349C / T366S / L368A / Y407V are introduced into CH3 of the other heavy chain (heavy chain B).

[0319] (2) A bispecific antibody in which an antibody fragment is fused to the C-terminus of the antibody.

[0320] (3) A bispecific antibody in which an antibody fragment is fused to the N-terminus of the antibody.

[0321] The bispecific antibody described in (1) may be a bispecific antibody in which the antigen-binding site comprising VH of heavy chain A binds to CADM3 and the antigen-binding site comprising VH of heavy chain B binds to an antigen present in the brain, or a bispecific antibody in which the antigen-binding sites bind in exactly opposite manners.

[0322] Examples of the bispecific antibodies described in (2) include a bispecific antibody in which an antibody fragment is linked to the C-terminus of one of the two heavy chains constituting the antibody, a bispecific antibody in which an antibody fragment is linked to the C-terminus of both of the two heavy chains constituting the antibody, a bispecific antibody in which an antibody fragment is linked to the C-terminus of one of the two light chains constituting the antibody, a bispecific antibody in which an antibody fragment is linked to the C-terminus of both of the two light chains constituting the antibody, a bispecific antibody in which an antibody fragment is linked to the C-terminus of each of the two light chains and the C-terminus of the two heavy chains constituting the antibody, etc. Note that a suitable linker may be present between the C-terminus of the antibody and the antibody fragment.

[0323] The antibody fragment contained in the bispecific antibody described in (2) is preferably scFv, Fab, VHH, etc., but is not particularly limited thereto.

[0324] The bispecific antibody described in (2) may be a bispecific antibody in which the antigen binding site at the N-terminus binds to CADM3 and in which the antigen binding site at the C-terminus binds to an antigen present in the brain or a bispecific antibody in which the antigen binding sites bind in exactly the opposite manner.

[0325] The bispecific antibody described in (3) is a bispecific antibody in which an antibody fragment is linked to the N-terminus of at least one of the two heavy chains or two light chains constituting the antibody. In addition, a suitable linker may be present between the N-terminus of the heavy chain and / or light chain of the antibody and the antibody fragment. The antibody fragment contained in the bispecific antibody described in (3) is preferably, but not particularly limited to, scFv, Fab, VHH, etc.

[0326] In addition, examples of the bispecific antibody described in (3) include a bispecific antibody having a VH1-CH1-VH2-CH1-hinge-CH2-CH3 structure from the N-terminus of the heavy chain, a bispecific antibody having the above-mentioned heavy chain structure in which VH1 and VH2 each form an antigen-binding site together with VL, etc. VL that forms an antigen-binding site with VH1 and VH2 may have the same amino acid sequence or different amino acid sequences.

[0327] In the present invention, the multispecific antibody or bispecific antibody may be any antibody, as long as it is a multispecific antibody or bispecific antibody that binds to CADM3. Among these antibodies, multispecific antibodies or bispecific antibodies that bind to CADM3 and an antigen present in the brain are preferred, and multispecific antibodies or bispecific antibodies comprising an antigen-binding site that binds to CADM3 and an antigen-binding site that binds to an antigen present in the brain are even more preferred.

[0328] In the present invention, examples of the antigen present in the brain include proteins, polysaccharides, lipids, etc., and among these antigens, the antigen is preferably a protein.

[0329] Examples of proteins present in the brain include prion protein, 5T4, AFP, ADAM10, ADAM12, ADAM17, AFP, AXL, BCAM, BSG, C5, C5R, CA9, CA72-4, CADM3, CCL11, CCL2, CCR1, CCR4, CCR5, CCR6, CD2, CD3E, CD4, CD5, CD6, CD8, CD11, CD18, CD19, CD20, CD22, CD24, CD25, CD29, 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, CDH2, CDH3, CEA, CEACAM1, CEACAM5, CEACAM6, CEACAM8, Claudin-3, Claudin-4, CSF-1, CSF2RA, CSPG-4, CSPG5, CTLA4, CRF-1, Cripto, CXCR4, CXCR5, DJ-1, DLL4, DR4, DR5, ED-B, EFNA2, E GFR, EGFRvIII, ETBR, ENPP3, EPCAM, EphA2, EphA4, EPOR, ERBB2, ERBB3, ERBB4, FAPα, FAS, FcγRI, FCER2, FGFR1, FGFR2, FGFR3, FGFR4, FLT1, FOLH1, FOLR1, GDF2, GFR, GLP1R, Glycan-3, GPNMB, GRP78, HAPLN4, HB-EGF, HGF, HLA-DRβ, HMGB1, ICAM1, ICAM5, IFNA1, IFNB, IgE, IgE-Fc, IGF1R, IL10, IL12B, I L13, IL15, IL17A, IL1A, IL1B, IL2RA, IL4, IL5, IL5RA, IL6, IL6R, IL9, IL2Rα, IL2Rβ, IL2Rγ, INSR, ITGA2, ITGA2B2, ITGB3, ITGA4, ITGB7, ITGA5, ITGAL, ITGAV, ITGB3, ITGB2, KDR, L1CAM, LAG3, LRP3, mesothelin, MAG, MMP14, MMP15, MOG, MST1R, MSTN, MUC1, MUC4, MUC16, MUC5AC, myostatin, connexin 4, NCAN, NGF, NMDAR,NOTCH, NRG1, NRP, OX40, OX40L, P2Y6, PAR1, PDGFA, PDGFB, PDGFRA, PDGFRB, PD1, PDL1, PLP1, PSCA, PTPRZ, RET, RGMA, SLAM7, SL C44A4, TAG-72, TCR, TGFB1, TGFB2, TGFBR, TIMP2, TLR9, TNF, TNFR, TNFRSF10A, TNFRSF10B, TNFRSF12A, TNFSF13, TNFSF14, TNFS F2, TNFSF7, TREM2, TRAILR2, TRKA, TRKB, TRKC, transferrin, VEGF, VEGFR, VLA-4, CGRP, α-synuclein, TDP-43, Tau, FUS, amyloid peptide-β (Aβ), APP, BACE1, presenilin, LINGO-1, Nogo, Troy, polyQ, androgen receptor, huntingtin protein, spinocerebellar ataxia protein 1, spinocerebellar ataxia protein 2, phosphorylated Tau, phosphorylated α-synuclein, etc., but the proteins are not limited to these proteins.

[0330] Examples of glycans present in the brain include Lewis-x, Lewis-y, CD15, and the like, but the glycans are not limited to these glycans.

[0331] Examples of lipids present in the brain include GD1a, GD2, GD3, GM1, GM2, GM3, phosphatidylserine, and the like, but the lipids are not limited to these lipids.

[0332] The antibodies or antibody fragments thereof of the present invention also include antibodies comprising any post-translationally modified amino acids. Examples of such post-translational modifications include deletion of lysine residues at the C-terminus of the H chain (lysine trimming), conversion of glutamine residues at the N-terminus of the polypeptide to pyroglutamine (pyroGlu), etc. [Beck et al., Analytical Chemistry, 85, 715-736 (2013)].

[0333] In the antibodies or antibody fragments thereof of the present invention, amino acid modifications of the Fc region may be performed. Examples of amino acid modifications of the Fc region include those for stabilizing the antibody or regulating its half-life in blood. Specific examples of amino acid modifications of the Fc region include those described in WO 2006 / 033386, WO 2006 / 075668, WO 2011 / 122011, WO 2009 / 125825, and the like.

[0334] The antibodies or antibody fragments thereof of the present invention also include fusion antibodies or fusion antibody fragments thereof modified by attaching a desired molecule to the antibody or antibody fragment thereof. The method for modifying the antibody is not particularly limited, and any method can be used as long as the method can modify the desired amino acid residues and glycans.

[0335] For example, examples are chemical modification using a chemical reaction [Introduction to Antibody Engineering, Chijinshokan Co., Ltd. (1994); Kolb et al., Angew Chem Int Ed Engl. 40. 2004-21, 2001], modification by genetic engineering technology in which a recombinant protein expression vector is introduced into a suitable host cell for expression using genetic recombination technology, and the like.

[0336] In the present invention, examples of molecules used to modify the antibody or antibody fragment thereof include hydrophilic polymers, amphiphilic polymers, functional molecules, etc. Examples of the hydrophilic polymer and amphiphilic polymer include polyoxyalkylene, molecules containing polyols or polysaccharides, etc.

[0337] In the present invention, when the antibody or antibody fragment thereof is modified with another molecule by chemical modification, examples of modification sites are the constant region of the antibody or antibody fragment, and specifically, a Cys residue or an SS bond site at the C-terminus is preferred. It is also possible to introduce a residue that can be chemically modified later at any position of the antibody or antibody fragment in advance by genetic engineering techniques.

[0338] Furthermore, when the antibody or antibody fragment thereof is directly modified with another molecule by genetic engineering techniques, as the modification site, the N-terminus or C-terminus of the light chain or heavy chain of the antibody or antibody fragment is exemplified.

[0339] Examples of the polyoxyalkylene include polyethylene glycol (PEG), polypropylene glycol, polypropylene glycol-ethylene glycol, and the like composed of a straight chain or a branched chain.

[0340] Examples of the molecule containing polyol or polysaccharide include linear or branched polysaccharides in which glucose is polymerized, such as amylose, dextran, pullulan, glycogen, etc. Furthermore, the molecule is not limited to homopolysaccharides but may be heteropolysaccharides.

[0341] The molecular weight of the molecule containing a hydrophilic polymer or an amphiphilic polymer is not particularly limited, but is preferably 100 Da or more, and preferably, for example, 100 Da to 100 kDa.

[0342] Examples of such functional molecules include antigen-binding molecules, fragments of antigen-binding molecules, drugs, bioactive peptides, bioactive proteins, nucleic acids, radiolabeled compounds, polysaccharides, lipids, fluorescent compounds, etc. Molecules that have bispecificity as a result of modification with functional molecules such as antigen-binding molecules are bispecific antibodies.

[0343] Examples of the antigen-binding molecule include antibodies, receptors, ligands, and the like.

[0344] The fragment of the antigen-binding molecule may be any fragment as long as the fragment is a fragment of the antigen-binding molecule and has antigen-binding activity.

[0345] Examples of the drug include anticancer agents such as alkylating agents, nitrosourea agents, antimetabolites, 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 and kinase inhibitors [Clinicaloncology, Japanese Journal of Cancer and Chemotherapy (1996)], anti-inflammatory agents such as steroidal agents, nonsteroidal agents, immunomodulators, immunosuppressants and antihistamines [Inflammation and anti-inflammatory therapy, Ishiyaku Publishers, Inc. (1982)] and the like.

[0346] More specific examples thereof include mertansine, emtansine, amifostine (Ethyol), cisplatin, dacarbazine (DTIC), dactinomycin, dichloromethyldiethylamine (nitrogen mustard), streptozotocin, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), doxorubicin (Adriamycin), epirubicin, gemcitabine (Gemzar), daunomycin, procarbazine, mitomycin, cytarabine, etoposide, 5-fluorouracil, fluorouracil, vinblastine, vincristine, bleomycin, daunomycin, peplomycin, estramustine, paclitaxel (Taxol), docetaxel, Taxotere, aldesleukin, asparaginase, busulfan, carboplatin, oxaliplatin, nedaplatin, cladribine, camptothecin, 10-hydroxy-7-ethyl-camptothecin (SN38), floxuridine, fludarabine, hydroxyurea, idarubicin, mesna, irinotecan (CPT-11), topotecan, mitoxantrone, topotecan, leuprorelin, megestrol acetate, melphalan, mercaptopurine, hydroxyurea, plicamycin, mitotane, pegaspargase, pentostatin, pipobroman, streptozotocin, tamoxifen, goserelin, leuprorelin, flutamide, teniposide, testolactone, thioguanine, thiotepa, uracil, vinorelbine, chlorambucil, hydrocortisone pine, 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 Tarceva, radicicol, 17-allylamino-17-demethoxygeldanamycin, rapamycin, amsacrine, all-trans retinoic acid, thalidomide, lenalidomide, anastrozole, fadrozole , letrozole, exemestane, bucillamine, azathioprine, mizoribine, cyclosporine, rapamycin, hydrocortisone, bexarotene (Targretin), tamoxifen, dexamethasone, progesterone, estrogen, anastrozole (Arimidex), leuprorelin, aspirin, indomethacin, celecoxib, azathioprine, penicillamine, gold thiomalate, chlorpheniramine maleate, chlorpheniramine, clematin, tretinoin, arsenic, bortezomib, allopurinol, calicheamicin, ibritumomab tiuxetan, targretin, ozogamicin, clarithromycin, tetrahydrofolate, ketoconazole, aminoglutethimide, suramin, methotrexate, maytansine alkaloids, etc., and their derivatives may also be included.

[0347] Examples of methods for linking the drug to the antibody or antibody fragment thereof include, in addition to the above methods, a method of linking the drug to an amino group of the antibody via glutaraldehyde, a method of linking the amino group of the drug to a carboxyl group of the antibody via water-soluble carbodiimide, and the like.

[0348] Examples of the biologically active peptides or biologically active proteins include interferon (IFN) -α, IFN-β, IFN-γ, interleukin (IL) -2, IL-12, IL-15, IL-18, IL-21, IL-23, granulocyte colony stimulating factor (G-CSF), granulocyte / macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), cytokines or growth factors that activate immune active cells such as NK cells, macrophages or neutrophils, proteases such as hydratases, lyases or isomerases, enzymes such as acid sphingomyelinase and glucocerebrosidase, toxins including bacterial toxins and plant toxins such as ricin, diphtheria toxin or ONTAK, antimicrobial peptides with cell membrane damaging activity, peptides with cell membrane affinity or cell membrane permeability, derivatives thereof, etc.

[0349] The nucleic acid may be any molecule as long as it is a molecule in which nucleotides or molecules having a function equivalent to that of nucleotides are polymerized, and examples thereof include siRNA, microRNA, antisense RNA / DNA, DNA aptamer, and the like.

[0350] The radioactive labeled compound may be any radioactive labeled compound as long as it is a nuclide used for diagnostic or therapeutic purposes, and examples thereof include 3 H. 14 C. 32 P. 33 P. 35 S. 51 Cr, 57 CO, 18 F. 153 Gd, 159 Gd, 64 Cu, 68 Ge, 166 Ho, 115 In, 113 In, 112 In, 111 In, 131 I. 125 I. 123 I. 121 I. 140 La, 177 Lu, 54 Mn, 99 Mo, 103 pd,142 pr, 149 pm, 186 Re、 188 Re、 211 At 105 Rh, 97 Such as 153 Sm, 47 Sc, 75 Se, 85 Sr. 99 Tc, 201 Ti, 113 Sn, 117 Sn, 133 Xe, 169 Yb, 175 Yb, 90 Y. 65 Zn, etc., or a compound containing any of said nuclides.

[0351] The radiolabeled compound can be directly linked to the antibody by the chloramine T method or the like. In addition, a substance that chelates the radiolabeled compound can be linked to the antibody. Examples of the chelating agent include 1,4,7,10-tetraazacyclododecanetetraacetic acid (DOTA), 1-[2-(4-aminophenyl)ethyl]-1,4,7,10-tetraazacyclododecanetetraacetic acid (PA-DOTA), 1,4,7,10-tetraazacyclotridecanetetraacetic acid (TRITA), diethylenetriaminepentaacetic acid (DTPA), and the like, and antibodies modified with the chelating agent and modified antibodies labeled with the radiolabeled compound via the chelating agent are also included in the antibodies of the present invention.

[0352] Examples of the polysaccharide include monosaccharides, disaccharides, and oligosaccharides, and more specific examples thereof include fucose, mannose, glucose, allose, aldose, gulose, idose, galactose, talose, ribose, arabinose, xylose, lyxose, erythose, erythrose, threose, cellobiose, maltose, isomaltose, lactose, lipoarabinomannan, Lewis X trisaccharide, sialyl Lewis X tetrasaccharide, etc. In addition, the polysaccharide may be a natural product containing a polysaccharide known as an immune adjuvant, examples of which include β (1→3) glucan (lentinan or schizophyllan), α-galactosylceramide (KRN7000), etc.

[0353] Examples of the lipids include simple lipids (neutral lipids), which are esters of fatty acids with any of various types of alcohols or their analogs. Examples include fats (e.g., triacylglycerols), waxes (e.g., fatty acid esters of long-chain alcohols), sterol esters, cholesterol esters, fatty acid esters of vitamins, and the like; complex lipids having polar groups such as phosphoric acid, sugar, sulfuric acid, or amines in addition to fatty acids and alcohols, such as phospholipids (e.g., glycerophospholipids, sphingomyelin, etc.) and glycolipids (e.g., glyceroglycolipids, sphingolipids, etc.); and derivative lipids, which refer to fat-soluble compounds such as fatty acids, long-chain alcohols, fat-soluble vitamins, steroids, sugars, and the like, among compounds produced by hydrolysis of simple lipids or complex lipids.

[0354] Examples of the fluorescent compound include fluorescent dyes, including fluorescein series such as fluorescein isothiocyanate (FITC), rhodamine series such as rhodamine isothiocyanate (RITC), Cy3, Cy5, eosin series, Alexa Fluor series, NBD series, etc., luminescent substances such as acridinium ester or lophan, fluorescent proteins such as green fluorescent protein (GFP), etc.

[0355] The hydrophilic polymer, amphiphilic polymer or functional molecule can be directly or via a suitable linker to the antibody or antibody fragment thereof of the present invention. Examples of the linker include esters, disulfides, hydrazones, dipeptides, and the like.

[0356] When a fusion antibody or fusion antibody fragment is produced by modifying an antibody of the present invention or an antibody fragment thereof using genetic engineering techniques, the fusion antibody or fusion antibody fragment can be produced as follows: cDNA encoding a protein is ligated to cDNA encoding an antibody to thereby construct a DNA encoding the fusion antibody or fusion antibody fragment, the DNA is inserted into an expression vector for prokaryotes or eukaryotes, the expression vector is introduced into a prokaryote or eukaryote, and the fusion antibody or fusion antibody fragment is expressed.

[0357] The composition of the present invention may be any composition, as long as the composition comprises the antibody of the present invention or its antibody fragment. In addition to the antibody or its antibody fragment, the composition may further comprise a suitable carrier or additive such as a stabilizer.

[0358] Examples of the composition of the present invention include compositions for detection or measurement comprising the antibody of the present invention or its antibody fragment, etc. Examples of the composition of the present invention include pharmaceutical compositions (therapeutic agents) comprising the antibody of the present invention or its antibody fragment as an active ingredient, etc., and the composition is formulated into a desired dosage form together with a pharmaceutically acceptable carrier.

[0359] In the present invention, the composition for detection or measurement may be any composition as long as the composition comprises the antibody of the present invention or its antibody fragment and can detect or measure an antigen that specifically binds to the antibody of the present invention or its antibody fragment. Examples of antigens that specifically bind to the antibody of the present invention or its antibody fragment include CADM3 or CADM3 and an antigen present in the brain.

[0360] The antibodies or antibody fragments thereof of the present invention, when administered to animals, have the property of binding to and accumulating CADM3 in the brain. Therefore, by using a composition for detection or measurement comprising the antibodies or antibody fragments thereof, the antibodies can be maintained in the brain, or the antibody concentration in the brain can be increased, allowing CADM3 or CADM3 and an antigen present in the brain to be detected or measured over a long period of time, and / or allowing CADM3 or CADM3 and an antigen present in the brain to be detected or measured with high sensitivity.

[0361] For example, when the composition for detection or measurement is a composition comprising a bispecific antibody that binds to CADM3 and an antigen present in the brain, CADM3 and the antigen present in the brain bound by the bispecific antibody can be detected or measured for a long time and / or can be detected or measured with high sensitivity.

[0362] Furthermore, for example, when the composition for detection or measurement is a composition comprising a fusion antibody or a fusion antibody fragment thereof that is labeled with a radiolabeled compound or a fluorescent dye and binds to CADM3, CADM3 can be detected or measured for a long time and / or can be detected or measured with high sensitivity.

[0363] The pharmaceutical composition (therapeutic agent) comprising the antibody of the present invention or its antibody fragment can be a therapeutic agent for any disease as long as the antigen to which the antibody of the present invention or its antibody fragment specifically binds is expressed in the disease, but is preferably a therapeutic agent for brain diseases.

[0364] Examples of such brain diseases include Alzheimer's disease, a prodromal stage of Alzheimer's disease, 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, spinal bulbar muscular atrophy, spinocerebellar degeneration, cerebrovascular disorders, epilepsy, migraine, attention deficit hyperactivity disorder, Creutzfeldt-Jakob disease, corticobasal degeneration, lysosomal storage diseases, depression, dystonia, and the like.

[0365] The antibodies or antibody fragments of the present invention, when administered to animals, have the property of binding to and accumulating CADM3 in the brain. Therefore, by using a therapeutic agent containing such an antibody or antibody fragment, the antibody or antibody fragment can be maintained in the brain for a long time, and the antibody concentration in the brain can be increased, thereby exhibiting a therapeutic effect on the aforementioned diseases.

[0366] For example, when the therapeutic agent is a fusion antibody comprising an anti-CADM3 antibody of the present invention, the therapeutic effect of the molecule can be exhibited by delivering the fusion molecule to the brain. Specifically, when the therapeutic agent is a fusion antibody comprising a drug, enzyme, or the like fused to an anti-CADM3 antibody, the therapeutic effect of the drug or enzyme can be exhibited, and when the therapeutic agent is a bispecific antibody that binds to CADM3 and an antigen present in the brain, the therapeutic effect for brain diseases associated with the antigen present in the brain and bound by the bispecific antibody can be exhibited.

[0367] Furthermore, for example, when the therapeutic agent is a fusion antibody or fusion antibody fragment modified with a low molecular weight drug that binds to CADM3, it can exhibit a therapeutic effect on brain diseases targeted by the low molecular weight drug. In this case, the therapeutic effect is preferably higher when the therapeutic agent of the present invention is used than when the low molecular weight drug is used alone.

[0368] The therapeutic agent comprising the antibody of the present invention or its antibody fragment may be a therapeutic agent comprising only the antibody or its antibody fragment as an active ingredient. However, in general, the therapeutic agent is ideally provided as a pharmaceutical preparation produced by mixing with one or more pharmaceutically acceptable carriers using any method known in the art of pharmaceutical technology.

[0369] As the route of administration, the route that is most effective for treatment is preferably used, and examples thereof include oral administration or parenteral administration such as oral, intratracheal, intrarectal, subcutaneous, intradermal, intramuscular, intracerebroventricular, intrathecal, intranasal, intraperitoneal, or intravenous administration, with intravenous or intracerebroventricular administration being particularly preferred. Examples of dosage forms include sprays, capsules, tablets, powders, granules, syrups, emulsions, suppositories, injections, ointments, tapes, and the like.

[0370] The dose or frequency of administration varies depending on the intended therapeutic effect, administration method, duration of treatment, age, body weight, etc., but is usually 10 μg / kg to 20 mg / kg per day for adults.

[0371] Furthermore, the present invention also encompasses methods for retaining antibodies in the brain, methods for enhancing the property of antibodies to accumulate in the brain, and methods for increasing the concentration (or amount) of antibodies in the brain, each of which uses the antibodies of the present invention or antibody fragments thereof.

[0372] In addition, the present invention also relates to a peptide that binds to CADM3, a nucleic acid comprising a nucleotide sequence encoding the peptide, a transformant cell comprising a vector containing the nucleic acid, a method for producing the peptide comprising culturing the transformant cell and collecting the peptide from the culture solution, a composition comprising the peptide, or a method for detecting or measuring an antigen present in the brain, a method for diagnosing or treating a brain disease, a method for enhancing the property of a peptide to accumulate in the brain, or a method for increasing the amount of the peptide in the brain, each of which uses the peptide or composition.

[0373] The peptides of the present invention include fusion peptides in which the peptides are modified.

[0374] For the definitions of various terms related to the peptide that binds to CADM3 etc., the same definitions as those described above for the antibody that binds to CADM3 etc. are used unless otherwise specified.

[0375] Hereinafter, methods for producing the antibodies of the present invention or antibody fragments thereof, methods for treating diseases, methods for diagnosing diseases, and the like will be specifically described.

[0376] 1. Methods for producing antibodies

[0377] (1) Preparation of antigens

[0378] CADM3 serving as an antigen or cells expressing CADM3 can be obtained by introducing an expression vector containing a cDNA encoding full-length CADM3 or a partial-length cDNA into Escherichia coli, yeast, insect cells, animal cells, etc. In addition, CADM3 can also be obtained by purifying CADM3 from various types of animal cell lines, animal cells, animal tissues, etc. in which CADM3 is expressed in large quantities.

[0379] In addition, the animal cell lines, animal cells, animal tissues, etc. can also be used as antigens. In addition, synthetic peptides having a partial sequence of CADM3 are prepared using chemical synthesis methods such as the Fmoc method or the tBoc method, and the synthetic peptides can also be used as antigens.

[0380] A known tag such as FLAG or His can be added to the C-terminus or N-terminus of CADM3 or a synthetic peptide having a partial sequence of CADM3.

[0381] CADM3 used in the present invention can be produced using the methods described in Molecular Cloning, A Laboratory Manual, 2nd edition (Cold Spring Harbor Laboratory Press (1989)), Current Protocols In Molecular Biology (John Wiley & Sons (1987-1997)), etc., for example, by expressing DNA encoding CADM3 in host cells by the following method.

[0382] First, a recombinant vector is produced by inserting a full-length cDNA containing a region encoding CADM3 downstream of a promoter into a suitable expression vector. DNA fragments having a suitable length and containing a region encoding a polypeptide, prepared based on the full-length cDNA, can be used in place of the full-length cDNA. Subsequently, a transformant producing the polypeptide can be obtained by introducing the resulting recombinant vector into a host cell suitable for the expression vector.

[0383] As the expression vector, any vector can be used, as long as it can replicate autonomously or can be integrated into the chromosome of the host cell to be used, and contains a suitable promoter at a position capable of transcribing the DNA encoding the polypeptide. As the host cell, any cell, such as a microorganism belonging to the genus Escherichia (Escherichia), such as Escherichia coli (E. coli), yeast, insect cells, animal cells, etc., can be used, as long as the target gene can be expressed.

[0384] When using prokaryotes such as Escherichia coli as host cells, the expression vector is preferably a vector that can autonomously replicate in prokaryotes and also comprises a promoter, a ribosome binding sequence, a DNA in the region comprising encoding mankind CADM3 and a transcription termination sequence. In addition, although the transcription termination sequence is not essential for the expression vector, the transcription termination sequence is preferably placed in the downstream close to the structural gene. In addition, the recombinant vector can comprise a gene that controls the promoter.

[0385] As the expression vector, a plasmid in which the distance between the Shine-Dalgarno sequence (also called SD sequence) as a ribosome binding sequence and the start codon is adjusted to an appropriate length (eg, 6 to 18 nucleotides) is preferably used.

[0386] Furthermore, in the nucleotide sequence of the DNA encoding CADM3, nucleotides may be substituted so that codons become optimal for expression in a host, and as a result, the production rate of the target CADM3 can be increased.

[0387] As the expression vector, any vector can be used as long as it can express its function in the host cell to be used, and examples thereof include pBTrp2, pBTac1, pBTac2 (manufactured by Roche Diagnostics K.K. above), pKK233-2 (manufactured by Pharmacia Corporation), pSE280 (manufactured by Invitrogen, Inc.), pGEMEX-1 (manufactured by Promega Corporation), pQE-8 (manufactured by QIAGEN, Inc.), pKYP10 (JP-A-S58-110600), pKYP200 [Agricultural Biological Chemistry, 48, 669 (1984)], pLSA1 [Agric. Biol. Chem., 53, 277 (1989)], pGEL1 [Proc. Natl. Acad. Sci. USA, 82, 4306 (1985)], pBluescript II SK(-) (manufactured by Stratagene Corporation), 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), JP-A-S60-221091], pGKA2 [prepared from Escherichia coli IGKA2 (FERM BP-6798), JP-A-S60-221091], pTerm2 (U.S. Patent No. 4,686,191, U.S. Patent No. 4,939,094 and U.S. Patent No. 160,735), pSupex, pUB110, pTP5, pC194, pEG400 [J. Bacteriol., 172, 2392 (1990)], pGEX (manufactured by Pharmacia Corporation), pET system (manufactured by Novagen, Inc.), pME18SFL3, etc.

[0388] As a promoter, any promoter can be used as long as it can show its function in the host cell to be used. Examples are promoters such as trp promoter (Ptrp), lac promoter, PL promoter, PR promoter or T7 promoter derived from Escherichia coli, bacteriophage, etc. In addition, examples are promoters such as artificially designed and modified promoters such as tandem promoters in which two Ptrp are connected in series, tac promoter, lacT7 promoter or let I promoter, etc.

[0389] 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, Escherichia coli DH5α, and the like.

[0390] As a method for introducing a recombinant vector into a host cell, any method can be used as long as it is a method for introducing DNA into a host cell to be used, and examples include a method using calcium ions [Proc. Natl. Acad. Sci. USA, 69, 2110 (1972); Gene, 17, 107 (1982); and Molecular & General Genetics, 168, 111 (1979)].

[0391] When animal cells are used as hosts, as the expression vector, any vector can be used as long as it can exhibit its function in animal cells, and examples thereof include pcDNAI, pCDM8 (manufactured by Funakoshi Co., Ltd.), pAGE 107 [JP-A-H3-22979; Cytotechnology, 3, 133 (1990)], pAS3-3 (JP-A-H2-227075), pCDM8 [Nature, 329, 840 (1987)], pcDNAI / Amp (manufactured by Invitrogen, Inc.), pcDNA3.1 (manufactured by Invitrogen, Inc.), pREP4 (manufactured by Invitrogen, Inc.), pAGE103 [J. Biochemistry, 101, 1307 (1987)], pAGE210, pME18SFL3, pKANTEX93 (WO97 / 10354), N5KG1val (U.S. Patent No. 6,001,358), INPEP4 (manufactured by Biogen-IDEC, Inc.), pCI (manufactured by Promega Corporation), transposon vector (WO 2010 / 143698) etc.

[0392] As the promoter, any promoter can be used as long as it can exhibit its function in animal cells, and examples thereof include cytomegalovirus (CMV) immediate early (IE) gene promoter, SV40 early promoter, retrovirus promoter, metallothionein promoter, heat shock promoter, SRα promoter, and Moloney murine leukemia virus promoter or enhancer. In addition, the human CMV IE gene enhancer can be used together with the promoter.

[0393] Examples of host cells include human leukemia cells Namalwa cells, monkey cells 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); Chromosoma, 41, 129 (1973); Methods in Cell Science, 18, 115 (1996); Radiation Research, 148, 260 (1997); Proc. Natl. Acad. Sci. USA, 77, 4216 (1980); Proc. Natl. Acad. Sci. USA, 60, 1275 (1968); Cell, 6, 121 (1975); Molecular Cell Genetics, 18, 115 (1996); Radiation Research, 148, 260 (1997); Proc. Natl. Acad. Sci. USA, 77, 4216 (1980); Proc. Natl. Acad. Sci. USA, 60, 1275 (1968); Cell, 6, 121 (1975); Molecular Cell Genetics, 18, 115 (1996); [Genetics, Appendix I, II (pp. 883-900)], dihydrofolate reductase gene (dhfr)-deficient CHO cells (CHO / DG44 cells) [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, catalog number 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 (JP-A-S63-000299), etc.

[0394] As a method for introducing an expression vector into a host cell, any method can be used as long as it is a method for introducing DNA into an animal cell. Examples thereof include electroporation [Cytotechnology, 3, 133 (1990)], calcium phosphate method (JP-A-H2-227075), lipofection method [Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)] and the like.

[0395] CADM3 can be produced by culturing a transformant derived from a microorganism, animal cell, or the like, obtained as described above, and harboring an expression vector incorporating a DNA encoding CADM3, in a culture medium to produce and accumulate CADM3 in the culture solution, and then collecting CADM3 from the culture solution. Culturing the transformant in a culture medium can be performed according to conventional methods for culturing a host.

[0396] When expressed in cells derived from eukaryotic organisms, CADM3 to which sugars or glycans are added can be obtained.

[0397] When culturing microorganisms transformed with an expression vector using an inducible promoter, an inducer may be added to the culture medium as needed. For example, when culturing microorganisms transformed with an expression vector using the lac promoter, isopropyl-β-D-thiogalactopyranoside or the like may be added to the culture medium, and when culturing microorganisms transformed with an expression vector using the trp promoter, indoleacrylic acid or the like may be added to the culture medium.

[0398] Examples of the culture medium for culturing transformants obtained using animal cells as hosts include commonly used RPMI1640 medium [The Journal of the American Medical Association, 199, 519 (1967)], Eagle's MEM medium [Science, 122, 501 (1952)], Dulbecco's modified MEM medium [Virology, 8, 396 (1959)], medium 199 [Proc. Soc. Exp. Biol. Med., 73, 1 (1950)], Iscove's modified Dulbecco's medium (IMDM), or any of these media supplemented with fetal bovine serum (FBS) or the like. The culture is usually performed at pH 6 to 8 and 30 to 40°C in the presence of 5% CO 2 for 1 to 7 days. In addition, during the culture period, antibiotics such as kanamycin or penicillin may be added to the medium as needed.

[0399] As a method for expressing a gene encoding CADM3, in addition to direct expression, examples include, for example, secretory production or fusion protein expression methods [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989)].

[0400] Examples of methods for producing CADM3 include methods of producing CADM3 in host cells, methods of secreting CADM3 outside host cells, and methods of producing CADM3 on the outer membrane of host cells, and a suitable method can be selected by changing the host cells to be used or the structure of CADM3 to be produced.

[0401] When CADM3 is produced in host cells or on the outer membrane of host cells, CADM3 can be actively secreted outside the host cells using the method of Paulson et al. [J. Biol. Chem., 264, 17619 (1989)], the method of Lowe et al. [Proc. Natl. Acad. Sci., USA, 86, 8227 (1989); Genes Develop., 4, 1288 (1990)], or the methods described in JP-A-H05-336963, WO 94 / 23021, etc. Furthermore, the production of CADM3 can be increased by utilizing a gene amplification system using the dihydrofolate reductase gene (JP-A-H2-227075), etc.

[0402] The obtained CADM3 can be isolated and purified, for example, as follows. When CADM3 is expressed in a dissolved state in cells, the cells are collected by centrifugation after completion of culture, suspended in an aqueous buffer solution, and then homogenized using an ultrasonic homogenizer, a French press, a Manton Gaulin homogenizer, a Dyno mill, or the like to obtain a cell-free extract solution. From the supernatant obtained by centrifugation of the cell-free extract solution, a purified preparation can be obtained using methods such as conventional protein separation and purification methods, i.e., solvent extraction, salting-out using ammonium sulfate or the like, desalting, precipitation using an organic solvent, anion exchange chromatography using a resin such as diethylaminoethyl (DEAE)-Sepharose or DIAION HPA-75 (manufactured by Mitsubishi Chemical Corporation), cation exchange chromatography using a resin such as S-Sepharose FF (manufactured by Pharmacia Corporation), hydrophobic chromatography using a resin such as butyl sepharose or phenyl sepharose, gel filtration using molecular sieves, affinity chromatography, chromatofocusing, electrophoresis such as isoelectric focusing electrophoresis, and the like, alone or in combination.

[0403] When CADM3 is expressed in cells by forming insoluble bodies, the cells are collected, homogenized in the same manner as described above, and then centrifuged to collect the insoluble CADM3 bodies as a sediment fraction. The collected insoluble CADM3 bodies are dissolved with a protein denaturant. CADM3 is returned to its normal conformation by dilution or dialysis, and a purified preparation of the polypeptide can then be obtained using the same isolation and purification methods as described above.

[0404] When CADM3 or its derivatives, such as glycomodified forms, are secreted extracellularly, CADM3 or its derivatives, such as glycomodified forms, can be collected in the culture supernatant. The culture can be treated in the same manner as described above using a method such as centrifugation to obtain a soluble fraction, and a purified preparation can be obtained from the soluble fraction using the same separation and purification methods as described above.

[0405] In addition, CADM3 used in the present invention can also be produced using a chemical synthesis method such as the Fmoc method or the tBoc method. In addition, chemical synthesis can also be performed using a peptide synthesizer manufactured by Advanced Chemtech, Inc., PerkinElmer, Inc., Pharmacia Corporation, Protein Technology Instrument, Inc., Synthecell-Vega Biomolecules Corporation, Perceptive, Inc., Shimadzu Corporation, etc.

[0406] (2) Animal immunization and preparation of antibody-producing cells for fusion

[0407] Animals aged 3 to 20 weeks, such as mice, rats, rabbits or hamsters, are immunized with the antigen obtained in (1), and antibody-producing cells are collected from the spleen, lymph nodes or peripheral blood of the animals. In addition, animals such as llamas, alpacas or camels can also be used as animals for immunization.

[0408] The immunization is carried out by administering the antigen to the animal subcutaneously, intravenously or intraperitoneally, for example, together with a suitable adjuvant such as Freund's complete adjuvant, aluminum hydroxide gel or Bordetella pertussis vaccine. When the antigen is a partial peptide, a conjugate of the antigen with a carrier protein such as bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) is produced and used as an immunogen.

[0409] When mice or rats are immunized, the antigen is administered 5 to 10 times every 1 to 2 weeks after the first administration. On the 3rd to 7th day after each administration, blood is collected from the venous plexus of the fundus of the eye, and the antibody titer of its serum is measured using an enzyme immunoassay [Antibodies-A Laboratory Manual, Cold Spring Harbor Laboratory (1988)]. Animals whose serum shows sufficient antibody titers against the antigen used for immunization are used as a supply source of antibody-producing cells for fusion.

[0410] On the 3rd to 7th day after the last administration of the antigen, tissues such as the spleen containing antibody-producing cells are removed from the immunized animal and the antibody-producing cells are collected. When splenocytes are used, the spleen is minced and dispersed, then centrifuged, and then the red blood cells are removed to obtain antibody-producing cells for fusion.

[0411] Other animals to be immunized can also be immunized in the same manner, and antibody-producing cells can be obtained. Suitable conditions for the time interval between immunizations and the length of time between the last immunization and tissue removal can be selected according to the animal species to be immunized.

[0412] (3) Preparation of myeloma cells

[0413] As myeloma cells, established cell lines obtained from mice are used, and for example, 8-azaguanine-resistant mouse (BALB / c-derived) myeloma cell line 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)], P3-X63-Ag8 (X63) [Nature, 256, 495 (1975)] and the like are used.

[0414] The myeloma cells were subcultured in a normal medium [RPM11640 medium supplemented with glutamine, 2-mercaptoethanol, gentamicin, FBS and 8-azaguanine] and then subcultured in a normal medium 3 to 4 days before cell fusion, and on the day of fusion, 2×10 7 More than cells.

[0415] (4) Cell fusion and preparation of hybridomas producing monoclonal antibodies

[0416] The antibody-producing cells for fusion obtained in (2) and the myeloma cells obtained in (3) are thoroughly washed and mixed with minimum essential medium (MEM) or phosphate-buffered saline (PBS: 1.83 g disodium hydrogen phosphate, 0.21 g potassium dihydrogen phosphate, 7.65 g sodium chloride, 1 L distilled water, pH 7.2) so that the cell number becomes as follows: antibody-producing cells for fusion: myeloma cells = 5:1 to 10:1, followed by centrifugation, and then the supernatant is removed.

[0417] After the precipitated cell aggregates were sufficiently dispersed, a mixed solution of polyethylene glycol 1000 (PEG-1000), MEM medium, and dimethyl sulfoxide was added thereto while stirring at 37° C. Furthermore, 1 to 2 mL of MEM medium was added thereto several times every 1 to 2 minutes, and then MEM medium was added thereto to make a total amount of 50 mL.

[0418] After centrifugation, the supernatant was removed. The precipitated cell aggregates were gently disrupted and then gently suspended in HAT medium (normal culture medium supplemented with hypoxanthine, thymidine, and aminopterin). The resulting suspension was cultured in a 5% CO2 incubator at 37°C for 7 to 14 days.

[0419] After cultivation, a portion of the culture supernatant is removed and cell aggregates that react with CADM3 but not with antigens other than CADM3 are selected using a hybridoma selection method, such as the binding assay described below. Subsequently, cloning is performed by limiting dilution, and cells that consistently produce high antibody titers are selected as monoclonal antibody-producing hybridomas.

[0420] (5) Preparation of purified monoclonal antibodies

[0421] The hybridoma producing the monoclonal antibody obtained in (4) was intraperitoneally injected into 8- to 10-week-old mice or nude mice that had been treated with pristane [0.5 mL of 2,6,10,14-tetramethylpentadecane (pristane) was intraperitoneally administered and then housed for 2 weeks]. Within 10 to 21 days, the hybridoma transformed into an ascites tumor.

[0422] Ascites is collected from the mouse, and then the solid is removed by centrifugation, followed by salting out using 40 to 50% ammonium sulfate. Subsequently, purification is performed by caprylic acid precipitation, a DEAE-agarose gel column, a protein A column, or a gel filtration column, and then the IgG or IgM fraction is collected to prepare a purified monoclonal antibody.

[0423] Furthermore, after culturing the monoclonal antibody-producing hybridoma obtained in (4) in RPMI1640 medium supplemented with 10% FBS, etc., the supernatant is removed by centrifugation, and the residue is suspended in hybridoma-SFM medium and then cultured for 3 to 7 days.

[0424] Purified monoclonal antibodies can also be obtained by centrifuging the obtained cell suspension, purifying the obtained supernatant by passing it through a protein A column or a protein G column, and then collecting the IgG fraction. Note that 5% Daigo's GF21 can also be added to the hybridoma-SFM medium.

[0425] Antibody subclassification was performed by enzyme immunoassay using a subclass typing kit. Protein quantification was performed by the Lowry method or by calculation from absorbance at 280 nm.

[0426] (6) Antibody selection

[0427] The selection of antibodies is performed as follows by measuring the affinity of the antibodies for CADM3-expressing cells using flow cytometry, etc. The CADM3-expressing cells may be any cells as long as CADM3 is expressed on the cell surface, and examples thereof include animal cells, animal cell lines, the CADM3-forced expression cell line obtained in (1), and the like.

[0428] After the CADM3-expressing cells are distributed into a plate such as a 96-well plate, a test substance such as serum, hybridoma culture supernatant, or purified antibody is distributed therein as the first antibody and allowed to react. After the reaction, the cells are thoroughly washed with PBS containing 1 to 10% BSA (hereinafter referred to as BSA-PBS) or the like, and then an anti-immunoglobulin antibody labeled with a fluorescent reagent or the like as the second antibody is distributed therein and allowed to react. After being thoroughly washed with BSA-PBS or the like, the fluorescence of the labeled antibody is measured using a flow cytometer, thereby selecting an antibody that specifically reacts with cells expressing CADM3.

[0429] Alternatively, antibodies can be selected by measuring the affinity of monoclonal antibodies for CADM3-expressing cells, CADM3 proteins, etc. using ELISA or surface plasmon resonance as described below. The CADM3 protein may be a protein composed of certain domains of CADM3 or a protein to which a tag such as GST is added.

[0430] In ELISA, after the CADM3-expressing cells or CADM3 protein are dispensed into a plate, such as a 96-well plate, the wells are blocked with BSA-PBS, and a test substance, such as serum, hybridoma culture supernatant, or purified antibody, is dispensed as the primary antibody and allowed to react. Subsequently, after sufficient washing with PBS or the like, an anti-immunoglobulin antibody labeled with a fluorescent reagent or the like is dispensed as the secondary antibody and allowed to react.

[0431] Then, after sufficient washing with PBS or the like, a color development solution is added. At the end, the color development reaction is terminated with a reaction termination solution, and the absorbance in each well is measured using a microplate reader to select antibodies that specifically react with the CADM3-expressing cells or CADM3 protein.

[0432] In surface plasmon resonance, the affinity of an antibody binding to CADM3 can be measured by immobilizing the antibody on a suitable sensor chip and using CADM3 protein as an analyte using a known protocol.

[0433] The obtained antibody affinity can be used to select an antibody having a desired affinity for the CADM3 protein. In addition, the affinity of an antibody binding to CADM3 can also be measured by immobilizing the CADM3 protein on a sensor chip and using the antibody as an analyte.

[0434] Furthermore, antibodies that compete with the antibodies of the present invention for binding to CADM3 can be obtained by adding the test antibody to the aforementioned flow cytometry or ELISA assay system to induce a reaction. In other words, by screening for antibodies that inhibit the binding of the antibodies of the present invention to CADM3 when the test antibody is added, antibodies that compete with the antibodies of the present invention for binding to the amino acid sequence or conformation of CADM3 can be obtained.

[0435] In addition, antibodies that bind to an epitope containing the epitope to which the antibody of the present invention binds can be obtained by identifying the epitope of the antibody obtained by the above-mentioned screening method by a known method, producing a synthetic peptide containing the identified epitope, a synthetic peptide manufactured to mimic the conformation of the epitope, etc., and then using it for immunization.

[0436] In addition, antibodies that bind to the same epitope as the epitope bound by the antibodies of the present invention can be obtained as follows: identify the epitope of the antibody obtained by the above-mentioned screening method, produce a partial synthetic peptide of the identified epitope, a synthetic peptide made to mimic the conformation of the epitope, etc., and then use it for immunization.

[0437] (7) Obtaining antibodies through phage display method

[0438] (7-1) Method for generating antibody phage library

[0439] In the present invention, as antibody phage libraries, immune libraries, natural libraries, and synthetic libraries can be used. The production methods of the corresponding libraries will be described below.

[0440] For the immune library, lymphocytes derived from animals or patients immunized in the same manner as described in (1) above are collected, and for the naive library, lymphocytes derived from normal animals or healthy humans are collected, RNA is extracted from the lymphocytes, and cDNA is synthesized by reverse transcription reaction.

[0441] The antibody gene fragment amplified by PCR using each cDNA as a template is inserted into a phagemid vector and transformed into Escherichia coli with the phagemid vector. When the transformant obtained is infected with a helper phage, an antibody phage library of the antibody gene can be obtained.

[0442] In addition, for synthetic libraries, the CDRs of the V genes in genomic DNA or reconstructed functional V genes are replaced with oligonucleotides encoding random amino acid sequences of appropriate lengths, and E. coli is transformed with a phagemid vector into which the V genes are inserted. When the resulting transformants are infected with helper phage, an antibody phage library can be obtained.

[0443] As the lymphocyte-derived cDNA and antibody phage libraries, commercially available products can also be used.

[0444] As the phagemid vector, pCANTAB 5E (Amersham Pharmacia Biotech, Inc.), pUC118 / pUC119 vector (TAKARA, Inc.), pBlueScript II phagemid vector (Agilent Technologies, Inc.), pKSTV-02 (Miyazaki et al., Biochem. 158 (3), 205-215, 2015), etc. can be used.

[0445] As the helper phage, M13KO7 helper phage (Invitrogen, Inc.), VCSM13 interference-resistant helper phage (Agilent Technologies, Inc.), R408 interference-resistant helper phage (Agilent Technologies, Inc.), etc. can be used.

[0446] Phage display can also use phage vectors. Examples include peptide phage libraries (manufactured by New England Biolabs, Inc., etc.) using filamentous phage g3p as a displayed molecule and methods using g7p, g8p, or g9p as a displayed molecule.

[0447] Phage display using T7 phage can also be used. Examples of display systems on T7 phage include the T7 Select vector (Novagen, Inc.).

[0448] (7-2) Selection of antibody phage clones

[0449] Selection of antibody phage clones from the antibody phage library generated in (7-1) can be performed using the ELISA method shown below.

[0450] CADM3 is immobilized on an immunotube and the tube is blocked with blocking buffer. The antibody phage library produced in (7-1) is added to each well of the tube and allowed to react. Subsequently, the wells are cleaned, fluorescently labeled anti-phage antibodies are added, and allowed to react. Subsequently, the wells are cleaned again and a color development solution is added. The color development reaction is then terminated with a reaction stop solution, and the absorbance in each well is measured using a microplate reader. In this way, antibody phage clones that bind to CADM3 are selected.

[0451] 2. Production of genetically recombinant antibodies

[0452] As examples of genetically recombinant antibody production, methods for producing human chimeric and humanized antibodies will be described below. Genetically recombinant mouse, rat, rabbit, hamster, camel, llama, alpaca, and human antibodies, as well as various types of chimeric antibodies and heavy chain antibodies, can also be produced in the same manner.

[0453] (1) Construction of expression vector for genetically recombinant antibodies

[0454] The expression vector for genetically recombinant antibodies is an expression vector for animal cells into which DNAs encoding CH and CL of human antibodies are incorporated, and can be constructed by cloning each of the DNAs encoding CH and CL of human antibodies into an expression vector for animal cells.

[0455] As the constant region (C region) of a human antibody, the CH and CL of any human antibody can be used. For example, the CH of the γ1 subtype and the CL of the κ subtype of a human antibody can be used. As DNA encoding the CH or CL of a human antibody, cDNA is used, but chromosomal DNA composed of exons and introns can also be used.

[0456] As the expression vector for animal cells, any vector can be used as long as it can incorporate a gene encoding the C region of a human antibody and express the gene. 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)], pSE1UK1Sed1-3 [Cytotechnol., 13, 79 (1993)], etc. can be used.

[0457] Examples of promoters and enhancers in the expression vector for animal cells include the SV40 early promoter [J. Biochem., 101, 1307 (1987)], Moloney murine leukemia virus LTR [Biochem. Biophys. Res. Commun., 149, 960 (1987)], or the immunoglobulin H chain promoter [Cell, 41, 479 (1985)] and enhancer [Cell, 33, 717 (1983)], etc.

[0458] As the expression vector for genetically recombinant antibodies, from the viewpoints of ease of construction of the expression vector for genetically recombinant antibodies, ease of introduction into animal cells, and balance of expression levels of the antibody H chain and L chain in the animal cells, an expression vector for genetically recombinant antibodies of a type (tandem type) in which the antibody H chain and L chain are present on the same vector is used [J. Immunol. Methods, 167, 271 (1994)], but a type in which the antibody H chain and L chain are present on separate vectors may also be used. As tandem type expression vectors for genetically recombinant antibodies, pKANTEX93 (WO 97 / 10354) and pEE18 [Hybridoma, 17, 559 (1998)] are used.

[0459] (2) Obtaining cDNA encoding the variable region (V region) of antibodies derived from animals other than humans and analyzing their amino acid sequences

[0460] Acquisition of cDNA encoding VH and VL of a non-human antibody and analysis of the amino acid sequence can be performed as follows.

[0461] (2-1) When antibodies are obtained by the hybridoma method

[0462] mRNA is extracted from non-human antibody-producing hybridoma cells and cDNA is synthesized. Each of the synthesized cDNAs is cloned into a vector such as a phage or a plasmid, thereby producing a cDNA library.

[0463] Using DNA encoding the C region domain or V region domain of a non-human antibody as a probe, a recombinant phage or recombinant plasmid containing each cDNA encoding VH and VL is isolated from the library. Each complete nucleotide sequence of the target VH or VL of the non-human antibody on the recombinant phage or recombinant plasmid is determined, and the complete amino acid sequence of each VH or VL is deduced from the nucleotide sequence.

[0464] As non-human animals used for producing hybridoma cells producing non-human antibodies, mice, rats, hamsters, rabbits, llamas, camels, alpacas, etc. are used, but any animal can be used as long as it can produce hybridoma cells.

[0465] For preparation of total RNA from hybridoma cells, the guanidine isothiocyanate-cesium trifluoroacetate method [Methods in Enzymol., 154, 3 (1987)] or a kit such as RNeasy kit (manufactured by QIAGEN, Inc.) or the like is used.

[0466] In the preparation of mRNA from total RNA, the oligo(dT)-immobilized cellulose column method [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989)] or a kit such as Oligo-dT30 <super>mRNA Purification (registered trademark) Kit (manufactured by Takara Bio, Inc.), etc. In addition, mRNA can also be prepared from hybridoma cells using a kit such as Fast Track mRNA Isolation (registered trademark) Kit (manufactured by Invitrogen, Inc.) or QuickPrep mRNA Purification (registered trademark) Kit (manufactured by Pharmacia Corporation).

[0467] In the synthesis of cDNA and the production of a cDNA library, a known method [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989), Current Protocols in Molecular Biology, Supplementary File 1, John Wiley & Sons (1987-1997)] or a kit such as the SuperScript Plasmid System for cDNA Synthesis and Plasmid Cloning (manufactured by Invitrogen, Inc.) or the ZAP-cDNA Synthesis (registered trademark) Kit (manufactured by Stratagene Corporation) or the like is used.

[0468] When generating a cDNA library, as a vector into which cDNA synthesized using mRNA extracted from hybridoma cells as a template is incorporated, any vector can be used as long as it is a vector capable of incorporating the cDNA. For example, ZAP ExPress [Strategies, 5, 58 (1992)], pBluescript II SK (+) [Nucleic Acids Research, 17, 9494 (1989)], λZAPII (manufactured by Stratagene Corporation), λgt10, λgt 11 [DNA Cloning: A Practical Approach, 1, 49 (1985)], Lambda Blue Mid (manufactured by Clontech Laboratories, Inc.), λExCell, pT7T3-18U (manufactured by Pharmacia Corporation), pCD2 [Mol. Cell. Biol., 3, 280 (1983)], pUC18 [Gene, 33, 103 (1985)] and the like are used.

[0469] As the Escherichia coli into which the cDNA library constructed by phage or plasmid vector is introduced, any Escherichia coli can be used as long as it can introduce, express and maintain the cDNA library. 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)], JM105 [Gene, 38, 275 (1985)] and the like are used.

[0470] To select a cDNA clone encoding VH or VL of a non-human antibody from the cDNA library, colony hybridization or plaque hybridization using an isotope- or fluorescent-labeled probe [Molecular Cloning, A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press (1989)] or the like is used.

[0471] Alternatively, cDNA encoding VH or VL can be prepared by preparing primers and performing a polymerase chain reaction (PCR) method using cDNA sy...

Claims

1. An antibody or an antibody fragment that binds to cell adhesion molecule 3 (CADM3), wherein the antibody or the antibody fragment comprises a variable heavy domain (VHH) of a heavy chain antibody, and wherein the VHH comprises complementarity-determining regions (CDR) 1 to 3 consisting of the amino acid sequences represented by SEQ ID NO: 13, 14, and 15, respectively.

2. The antibody or the antibody fragment according to claim 1, wherein the antibody has the property of accumulating in the brain.

3. The antibody or the antibody fragment according to claim 2, wherein the antibody has an affinity for neurons and / or nerve tissue.

4. The antibody or the antibody fragment according to claim 1, wherein the amino acid sequence of the VHH consists of the amino acid sequence represented by SEQ ID NO:

12.

5. The antibody or the antibody fragment according to claim 1, wherein the antibody or the antibody fragment is an antibody fragment, and wherein CDR1 to CDR3 of the VHH consist of the amino acid sequences represented by SEQ ID NO: 13, 14, and 15, respectively.

6. The antibody or the antibody fragment according to claim 5, wherein the antibody or the antibody fragment is an antibody fragment, and wherein the amino acid sequence of the VHH consists of the amino acid sequence represented by SEQ ID NO:

12.

7. The antibody or the antibody fragment according to any one of claims 1 to 6, wherein the antibody is a bispecific antibody.

8. The antibody or the antibody fragment according to claim 7, wherein the bispecific antibody comprises an antigen-binding site that binds to CADM3 and an antigen-binding site that binds to an antigen present in the brain.

9. The antibody or the antibody fragment according to any one of claims 1 to 6, wherein the antibody fragment is selected from Fab, Fab’, F(ab’)2, single-chain antibody (scFv), diabody, disulfide-stabilized V region (dsFv), and VHH.

10. The antibody or the antibody fragment according to any one of claims 1 to 6, wherein the antibody is a genetically recombinant antibody.

11. The antibody or the antibody fragment according to any one of claims 1 to 6, wherein the antibody is selected from mouse antibody, rat antibody, rabbit antibody, alpaca antibody, camel antibody, llama antibody, chimeric antibody, humanized antibody, and human antibody.

12. A fusion antibody or a fusion antibody fragment, wherein at least one selected from the following (i) to (iii) is linked to the antibody or the antibody fragment according to any one of claims 1 to 11: (i) A hydrophilic polymer; (ii) An amphiphilic polymer; and (iii) A functional molecule.

13. A nucleic acid that comprises a nucleotide sequence encoding the antibody or the antibody fragment according to any one of claims 1 to 11 or the fusion antibody or the fusion antibody fragment according to claim 12.

14. A transformed somatic cell that comprises a vector containing the nucleic acid according to claim 13.

15. A method for producing an antibody or an antibody fragment according to any one of claims 1 to 11, or a fusion antibody or a fusion antibody fragment according to claim 12, the method comprising: culturing the transformed somatic cells according to claim 14, and collecting from the culture solution an antibody or an antibody fragment according to any one of claims 1 to 11, or a fusion antibody or a fusion antibody fragment according to claim 12.

16. A composition comprising an antibody or an antibody fragment according to any one of claims 1 to 11, or a fusion antibody or a fusion antibody fragment according to claim 12.

17. The composition according to claim 16, which is a composition for detecting or measuring an antigen present in the brain.

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