Novel anti-L1CAM antibody

The screening of high specificity and high affinity anti-L1CAM monoclonal antibodies was obtained through hybridoma technology, which solved the problem of lack of effective therapeutic antibodies in the prior art, and achieved specific binding and efficient treatment of L1CAM.

CN119798440APending Publication Date: 2025-04-11CURON BIOPHARMACEUTICAL (SHANGHAI) CO LTD
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Patent Information

Application Number
CN202411997153.8
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Priority Date
2022-01-10
Filing Date
2022-11-01
Publication Date
2025-04-11

AI Technical Summary

Technical Problem

The lack of effective therapeutic antibodies against L1 cell adhesion molecules (L1CAM) in the prior art has led to increased difficulty in treating in a variety of tumors.

Method used

Anti-L1CAM monoclonal antibodies with high specificity, high affinity and high stability were obtained through hybridoma technology screening, and corresponding nucleic acids, plasmids, host cells and pharmaceutical compositions were constructed.

Benefits of technology

A fully human monoclonal antibody specifically binding to L1CAM can be used clinically to diagnose or treat autoimmune diseases, chronic viral infections and tumors, improving the therapeutic effect of L1CAM.

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Abstract

The invention provides a fully human monoclonal antibody combined with human L1CAM with high specificity, an antigen binding fragment of the fully human monoclonal antibody, a bispecific antibody of the fully human monoclonal antibody and the like. The antibodies of the invention can be used in the diagnosis and treatment of cancer.
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Description

[0001] This application is a divisional application of Chinese invention patent application No. 202280073016.8, with the filing date of November 10, 2022 and the invention name being “Novel anti-L1CAM antibody”.

[0002] This application claims priority to the Chinese patent application filed with the Chinese Patent Office on November 3, 2021, with application number 202111296133.4 and invention name “Novel anti-L1CAM antibody” and the Chinese patent application filed with the Chinese Patent Office on January 10, 2022, with application number 202210022509.0 and invention name “Novel anti-L1CAM antibody”, the entire contents of which are incorporated by reference into this application. Technical Field

[0003] The present invention belongs to the field of biotechnology, and specifically relates to a fully human monoclonal antibody, an antigen binding fragment and a chimeric antigen receptor that specifically bind to human L1CAM, as well as nucleic acids, plasmids, host cells required for constructing the antibody, antigen binding fragment and chimeric antigen receptor, and corresponding pharmaceutical compositions and applications thereof. Background Art

[0004] L1 cell adhesion molecule, also known as CD171, is a transmembrane glycoprotein with a molecular weight between 200,000 and 220,000 [Miriam van der Maten et al., Int. J. Mol. Sci., 2019, 20: 4180.]. The expression of L1 cell adhesion molecule in healthy people is mainly limited to the nervous system, and is expressed in small amounts in kidney tissue and skin [Annette Kunkele et al., Clin. Cancer Res., 2016, 23: 466-477.]. L1 cell adhesion molecule has a high expression level in many tumors and is closely related to the proliferation and drug resistance of many tumor cells.

[0005] L1 cell adhesion molecule is a highly potential target for tumor therapy, but a therapeutic antibody against L1 cell adhesion molecule is still needed. Summary of the invention

[0006] The inventors of the present invention used hybridoma technology to screen and obtain an anti-L1CAM monoclonal antibody with high specificity, high affinity and high stability.

[0007] In a first aspect, the present invention provides a L1CAM binding molecule, which includes a fully human immunoglobulin that specifically binds to L1CAM.

[0008] In another aspect, the present invention relates to nucleic acid molecules encoding L1CAM binding molecules, and expression vectors and host cells containing the nucleic acid molecules.

[0009] In addition, the present invention also discloses nucleic acids encoding these whole clone antibodies and antigen binding fragments. In some embodiments, vectors comprising these nucleic acids and host cells comprising these vectors are disclosed. In further embodiments, pharmaceutical compositions comprising these whole clone antibodies, antigen binding fragments, bispecific antibodies, immunoconjugates, fusion proteins, nucleic acids, and vectors are disclosed. In other embodiments, these whole clone antibodies, antigen binding fragments, bispecific antibodies, immunoconjugates, fusion proteins, nucleic acids, and vectors are used to diagnose or treat diseases such as autoimmune diseases, chronic viral infections, and tumors.

[0010] The present invention discloses a novel fully human monoclonal antibody specific to L1CAM. Using the antibody CDR region or part or the whole gene, different forms of genetically engineered antibodies can be transformed and produced in prokaryotic and eukaryotic cells and any expression system, and can be used to clinically diagnose or treat autoimmune diseases, chronic viral infections, tumors and other diseases.

[0011] The present invention discloses a fully human monoclonal antibody that specifically binds to human L1CAM, and a CDR region of a light chain variable region and / or a heavy chain variable region thereof. In some embodiments, the amino acid sequence of the light chain variable region and / or the amino acid sequence of the heavy chain variable region of the monoclonal antibody or antigen-binding fragment contains at least one CDR region of a light chain and / or heavy chain variable region. The present invention also discloses antigen-binding fragments of these monoclonal antibodies, bispecific antibodies, chimeric antigen receptor CARs, antibody conjugates, corresponding nucleic acids, plasmids and host cells, corresponding pharmaceutical compositions, and uses in the diagnosis and treatment of cancer.

[0012] The present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to L1CAM, which comprises a light chain variable region having light chain complementary determining regions LCDR1, LCDR2 and LCDR3 and / or a heavy chain variable region having heavy chain complementary determining regions HCDR1, HCDR2 and HCDR3, wherein the antibody or the antigen-binding fragment thereof can specifically bind to the Ig1 or FN-4 domain of L1-CAM and does not have a CDR combination that is identical or equivalent to that of antibody L1-9.3 or UJ127.

[0013] In the technical scheme of the present invention, the heavy chain variable region has HCDR1, HCDR2 and HCDR3 that are at least 80%, 90% or 100% identical to HCDR1, HCDR2 and HCDR3 of a heavy chain variable region sequence selected from SEQ ID NO:43, 45, 47, 49 and 51, and the light chain variable region has LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of a light chain variable region sequence selected from SEQ ID NO:44, 46, 48, 50 and 52.

[0014] In the technical solution of the present invention, it comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region have a CDR combination selected from the following:

[0015] 1) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:43, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:44;

[0016] 2) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:45, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:46;

[0017] 3) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:47, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:48;

[0018] 4) HCDR1, HCDR2 and HCDR3 having at least 80% identity with HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:49, and LCDR1, LCDR2 and LCDR3 having at least 80% identity with LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:50; and

[0019] 5) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:51, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:52.

[0020] In the technical solution of the present invention,

[0021] The HCDR1 has at least 80% identity to a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 13, SEQ ID NO: 19, SEQ ID NO: 25, SEQ ID NO: 31, SEQ ID NO: 37;

[0022] The HCDR2 has at least 80% identity to a sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 8, SEQ ID NO: 14, SEQ ID NO: 20, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 38;

[0023] The HCDR3 has at least 80% identity to a sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 9, SEQ ID NO: 15, SEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 39;

[0024] The LCDR1 has at least 80% identity to a sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:16, SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:34, SEQ ID NO:40;

[0025] The LCDR2 has at least 80% identity to a sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:23, SEQ ID NO:29, SEQ ID NO:35, SEQ ID NO:41;

[0026] The LCDR3 has at least 80% identity to a sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:42.

[0027] In the technical solution of the present invention, wherein:

[0028] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 1;

[0029] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 2;

[0030] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 3;

[0031] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 4;

[0032] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 5;

[0033] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 6.

[0034] In the technical solution of the present invention, wherein:

[0035] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 7;

[0036] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 8;

[0037] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 9;

[0038] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 10;

[0039] (5) The LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 11; (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 12. In the technical solution of the present invention, wherein:

[0040] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 13;

[0041] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 14;

[0042] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 15;

[0043] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 16; (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 17; (6) the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 18. In the technical solution of the present invention, wherein:

[0044] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 19;

[0045] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 20;

[0046] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 21;

[0047] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 22; (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 23; (6) the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 24. In the technical solution of the present invention, wherein:

[0048] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 25;

[0049] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 26;

[0050] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 27;

[0051] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 28; (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 29; (6) the LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 30. In the technical solution of the present invention, wherein:

[0052] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 31;

[0053] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 32;

[0054] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 33;

[0055] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 34; (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 35;

[0056] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 36.

[0057] In the technical solution of the present invention, wherein:

[0058] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 37;

[0059] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 38;

[0060] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 39;

[0061] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 40;

[0062] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 41;

[0063] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 42.

[0064] In the technical solution of the present invention, the heavy chain variable region comprises an amino acid sequence shown in SEQ ID NO: 43 or an amino acid sequence having at least 90%, at least 95% or 100% sequence identity with SEQ ID NO: 43.

[0065] In the technical solution of the present invention, the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 44 or an amino acid sequence having at least 90%, at least 95% or 100% sequence identity with SEQ ID NO: 44.

[0066] In the technical solution of the present invention, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 45 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 45.

[0067] In the technical solution of the present invention, the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 46 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 46.

[0068] In the technical solution of the present invention, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 47 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 47.

[0069] In the technical solution of the present invention, the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 48 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 48.

[0070] In the technical solution of the present invention, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 49 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 49.

[0071] In the technical solution of the present invention, the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 50 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 50.

[0072] In the technical solution of the present invention, the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 51 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 51.

[0073] In the technical solution of the present invention, the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 52 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 52.

[0074] In the technical solution of the present invention, the antibody is a humanized antibody.

[0075] The present invention provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 53 or SEQ ID NO: 54 or SEQ ID NO: 55 or SEQ ID NO: 56, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 57 or SEQ ID NO: 58 or SEQ ID NO: 59.

[0076] The present invention provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and / or a light chain variable region, wherein:

[0077] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0078] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0079] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59; or

[0080] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0081] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0082] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59; or

[0083] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0084] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0085] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59; or

[0086] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0087] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0088] 12) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59.

[0089] The present invention provides an antibody or an antigen-binding fragment thereof, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 60 or SEQ ID NO: 61 or SEQ ID NO: 62 or SEQ ID NO: 63, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 64 or SEQ ID NO: 65 or SEQ ID NO: 66.

[0090] The present invention provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and / or a light chain variable region, wherein:

[0091] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0092] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0093] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66; or

[0094] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0095] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0096] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66; or

[0097] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0098] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0099] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66; or

[0100] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0101] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0102] 12) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66.

[0103] The present invention provides an antibody or an antigen-binding fragment thereof, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 67 or SEQ ID NO: 68 or SEQ ID NO: 69 or SEQ ID NO: 70 or SEQ ID NO: 71 or SEQ ID NO: 72 or SEQ ID NO: 73 or SEQ ID NO: 74, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 75 or SEQ ID NO: 76 or SEQ ID NO: 77 or SEQ ID NO: 78 or SEQ ID NO: 79 or SEQ ID NO: 80 or SEQ ID NO: 81.

[0104] The present invention provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and / or a light chain variable region, wherein:

[0105] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0106] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0107] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0108] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0109] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0110] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0111] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0112] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0113] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0114] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0115] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0116] 12) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0117] 13) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0118] 14) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0119] 15) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0120] 16) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0121] 17) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0122] 18) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0123] 19) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0124] 20) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0125] 21) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0126] 22) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0127] 23) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0128] 24) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0129] 25) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0130] 26) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0131] 27) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0132] 28) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0133] 29) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0134] 30) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0135] 31) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0136] 32) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0137] 33) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0138] 34) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0139] 35) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0140] 36) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0141] 37) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0142] 38) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0143] 39) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0144] 40) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0145] 41) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0146] 42) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0147] 43) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0148] 44) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0149] 45) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0150] 46) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0151] 47) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0152] 48) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0153] 49) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0154] 50) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0155] 51) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0156] 52) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0157] 53) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0158] 54) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0159] 55) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0160] 56) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81.

[0161] The present invention provides an antibody or an antigen-binding fragment thereof, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 82 or SEQ ID NO: 83 or SEQ ID NO: 84 or SEQ ID NO: 85 or SEQ ID NO: 86 or SEQ ID NO: 87 or SEQ ID NO: 88 or SEQ ID NO: 89 or SEQ ID NO: 90, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 91 or SEQ ID NO: 92 or SEQ ID NO: 93 or SEQ ID NO: 94 or SEQ ID NO: 95 or SEQ ID NO: 96.

[0162] The present invention provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and / or a light chain variable region, wherein:

[0163] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0164] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0165] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0166] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0167] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0168] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0169] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0170] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0171] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0172] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0173] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0174] 12) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0175] 13) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0176] 14) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0177] 15) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0178] 16) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0179] 17) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0180] 18) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0181] 19) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0182] 20) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0183] 21) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0184] 22) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0185] 23) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0186] 24) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0187] 25) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0188] 26) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0189] 27) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0190] 28) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0191] 29) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0192] 30) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0193] 31) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0194] 32) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0195] 33) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0196] 34) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0197] 35) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0198] 36) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0199] 37) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0200] 38) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0201] 39) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0202] 40) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0203] 41) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0204] 42) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0205] 43) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0206] 44) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0207] 45) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0208] 46) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0209] 47) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0210] 48) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0211] 49) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0212] 50) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0213] 51) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0214] 52) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0215] 53) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0216] 54) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96.

[0217] Furthermore, the present invention provides a fully humanized antibody or an antigen-binding fragment thereof, which further comprises an Fc region, optionally comprising a human immune protein (Ig) Fc region, or optionally comprising the Fc region of human IgG, and the Fc region is derived from human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM.

[0218] The antigen-binding fragment of the present invention is scFv, scFv-Fc, Fv, Fab, Fab', F(ab)2 or F(ab')2.

[0219] Preferably, the antigen-binding fragment of the present invention is F(ab)2, Fab and scFv.

[0220] F(ab)2 fragments are "bivalent". "Bivalent" means that the antibody and F(ab')2 fragments have two antigen binding sites. In contrast, Fab, Fv, and scFv fragments are monovalent and have only one antigen binding site.

[0221] F(ab)2 is an antibody fragment having a molecular weight of about 50,000 and antigen-binding activity, among the fragments obtained by treating IgG antibody molecules with the protease papain (cleaving the amino acid residue at position 224 of the H chain), in which about half of the N-terminal side and the entire L chain are bound together by a disulfide bond (CAK Borrebaeck, editor (1995) Antibody Engineering (Breakthroughs in Molecular Biology), Oxford University Press; R. Kontermann & S. Duebel, editors (2001) Antibody Engineering (Springer Laboratory Manual), Springer Verlag).

[0222] Fab is an antibody fragment having a molecular weight of about 50,000 and antigen-binding activity obtained by cleaving the disulfide bond of the hinge region of the above-mentioned F(ab')2. The Fab of the present invention can be produced by treating F(ab)2 with a reducing agent such as dithiothreitol.

[0223] "Single-chain Fv" or "scFv" means a molecule comprising an antibody heavy chain variable domain (or region; VH) and an antibody light chain variable domain (or region; VL) connected by a linker. Such scFv molecules may have the general structure: H2-VL-linker-VH-COOH or H2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeated GGGGS amino acid sequences or variants thereof, for example using 1-4 repeated variants.

[0224] The antibody or antigen-binding fragment thereof described in the present invention is a monoclonal antibody, a bispecific antibody, a multispecific antibody, a recombinant antibody, a chimeric antibody, a bivalent antibody, an anti-idiotypic antibody or a fusion protein.

[0225] In addition, the present invention also provides an antibody-drug conjugate, comprising any of the aforementioned antibodies or antigen-binding fragments of the present invention, and an additional effector molecule conjugate.

[0226] In addition, the present invention also provides a nucleic acid molecule encoding any of the aforementioned antibodies or antigen-binding fragments of the present invention.

[0227] In the nucleic acid molecule of the present invention, it is operably linked to a promoter.

[0228] In addition, the present invention also provides a recombinant expression vector, which contains the aforementioned nucleic acid molecule of the present invention.

[0229] In addition, the present invention also provides a plasmid containing the nucleic acid molecule of the present invention.

[0230] In addition, the present invention also provides a host cell containing the nucleic acid molecule of the present invention, the recombinant expression vector of the present invention or the plasmid of the present invention.

[0231] In addition, the present invention also provides a method for detecting L1CAM expression in a biological sample, comprising the following steps: contacting the biological sample with any one of the antibodies or antigen-binding fragments thereof described above, and detecting the presence or absence of immune complexes, wherein the presence of immune complexes indicates the presence of L1CAM in the biological sample.

[0232] In addition, the present invention also provides a pharmaceutical composition, comprising any one of the aforementioned antibodies or antigen-binding fragments thereof of the present invention, the antibody-drug conjugate of the present invention as an active substance, and a pharmaceutically acceptable carrier.

[0233] The present invention provides a pharmaceutical composition, which is prepared by mixing an effective preventive or therapeutic dose of the antibody or antigen-binding fragment or bispecific antibody or chimeric antigen receptor or its nucleic acid molecule, plasmid or cell expressing the chimeric antigen receptor of the present invention with a physiologically or pharmaceutically acceptable carrier, excipient or stabilizer, and the composition includes but is not limited to a lyophilized dosage form, an aqueous solution dosage form, a liposome or a capsule dosage form, etc. The concentration of the monoclonal antibody or antigen-binding fragment or bispecific antibody or chimeric antigen receptor or its nucleic acid molecule, plasmid or cell expressing the chimeric antigen receptor of the present invention can be varied from about 0.1% to 100% (by weight).

[0234] In addition, the present invention further provides a kit, comprising any one of the aforementioned antibodies or antigen-binding fragments thereof of the present invention, the antibody-drug conjugate of the present invention and / or the pharmaceutical composition of the present invention.

[0235] In addition, the present invention also provides a method for treating cancer, which comprises administering any of the aforementioned antibodies or antigen-binding fragments thereof, the antibody-drug conjugates of the present invention, or the pharmaceutical composition of the present invention to a subject in need thereof.

[0236] In the above method of the present invention, it further comprises detecting the presence of L1-CAM in the cancer sample before administering the drug.

[0237] Preferably, the antibody or antigen-binding fragment of the present invention is fully human. The fully human refers to an antibody or antigen-binding fragment thereof defined herein as non-chimeric (e.g., non-"humanized") and not from (whether in whole or in part) a non-human species. A fully human antibody or antigen-binding fragment thereof may be derived from a human, or may be a synthetic human antibody. Another example of a fully human antibody or antigen-binding fragment thereof is a human antibody encoded by a nucleic acid isolated from a library of antibody sequences of human origin (e.g., such a library based on antibodies taken from natural sources of humans).

[0238] The antibody or antigen-binding fragment thereof of the present invention is a monoclonal antibody, a bispecific antibody, a multispecific antibody, a recombinant antibody, a chimeric antibody, a bivalent antibody, an anti-idiotypic antibody or a fusion protein.

[0239] The present invention also provides an antibody or antigen binding fragment capable of binding to L1CAM, which is a bispecific antibody or bispecific antigen binding fragment. In some embodiments, the bispecific antibody or bispecific antigen binding fragment can be separated. The term "bispecific" refers to an antigen binding molecule that can specifically bind to at least two different antigenic determinants. In some embodiments, the bispecific antibody and bispecific antigen binding fragment comprise an antigen binding fragment according to the present invention. In some embodiments, the bispecific antibody and bispecific antigen binding fragment comprise an antigen binding fragment capable of binding to L1CAM, wherein the antigen binding fragment capable of binding to L1CAM comprises an antigen binding fragment according to the present invention. In some embodiments, the bispecific antibody and bispecific antigen binding fragment comprise an antigen binding fragment capable of binding to L1CAM, and an antigen binding fragment capable of binding to another target protein. In some embodiments, the bispecific antibody or antigen binding fragment comprises an antigen binding molecule capable of binding to L1CAM and an antigen binding molecule capable of binding to other than L1CAM. In some embodiments, antigens other than L1CAM are immune cell surface molecules. In some embodiments, antigens other than L1CAM are cancer cell antigens. In some embodiments, antigens other than L1CAM are receptor molecules, such as cell surface receptors.

[0240] The present invention provides nucleic acids encoding the monoclonal antibody or antigen binding fragment or bispecific antibody or chimeric antigen receptor. The present invention provides plasmids comprising the nucleic acid, optionally operably connected to regulatory sequences, such as promoters, enhancers, etc. The present invention provides host cells comprising the plasmid, and methods for producing and optionally recovering the monoclonal antibody or antigen binding fragment or bispecific antibody or chimeric antigen receptor. The host cell of the present invention can be any prokaryotic cell or eukaryotic cell, including but not limited to bacterial cells (e.g., Escherichia coli, Bacillus subtilis), insect cells (e.g., using baculovirus expression systems), yeast or mammalian cells (e.g., CHO or BHK cell lines). Other suitable host cells are known to those skilled in the art.

[0241] Preferably, the disease of the present invention is cancer, including but not limited to lymphoma, blastoma, sarcoma (including liposarcoma), neuroendocrine tumor, mesothelioma, schwannoma, meningioma, adenoma, melanoma and non-leukemic leukemia or lymphoid malignancy. More specific examples of the above-mentioned cancer include squamous cell carcinoma (e.g., squamous epithelial cell carcinoma), lung cancer, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma and lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal cancer, pancreatic cancer, malignant glioma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penis cancer, testicular cancer, esophageal cancer, bile duct tumor, head cancer, neck cancer, bone marrow stromal tumor, osteoclastoma, multiple myeloma, osteolytic bone marrow cancer (osteolytic bone cancers), central nervous system tumors, brain tumors (glioma, neuroblastoma, astrocytoma, medulloblastoma, ependymoma, and retinoblastoma), nasopharyngeal carcinoma, basal cell carcinoma, bile duct carcinoma, Kaposi's sarcoma, primary liver or endometrial cancer, and vascular system tumors (angiosarcoma and hemagiopericytoma), hematological tumors, Hodgkin's lymphoma, non-Hodgkin's lymphoma (Burkitt's lymphoma, small lymphocytic lymphoma / chronic lymphocytic leukemia, mycosis fungoides, mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, marginal zone lymphoma, hairy cell leukemia, and lymphoplasmacytic leukemia), lymphocyte precursor tumors, B-cell acute lymphoblastic leukemia The following are examples of the following: peripheral T-cell non-leukemic leukemia / lymphoma, T-cell acute lymphoblastic non-leukemic leukemia / lymphoma, thymoma, mature T and NK cell neoplasms, peripheral T-cell non-leukemic leukemia, mature T-cell non-leukemic leukemia / T-cell lymphoma, large granular lymphocytic leukemia, Langerhans cell histiocytosis, myeloma of acute myeloid leukemia, mature acute myeloid leukemia (AML), differentiated acute myeloid leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, myelodysplastic syndrome, chronic myeloproliferative disease, chronic myeloid leukemia, hematological malignancies such as multiple myeloma (MM), myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML).

[0242] The antibodies or antigen-binding fragments or bispecific antibodies or chimeric antigen receptors or nucleic acid molecules thereof, plasmids or cells expressing the chimeric antigen receptors, and pharmaceutical compositions of the present invention can be administered to human or animal subjects via various routes of administration, which usually depend on the characteristics of the disease to be treated. In general, the methods of the present invention can be implemented using any medically acceptable mode of administration, including oral, rectal, topical, intraocular, intracisternal, intraventricular, intratracheal, intranasal instillation, transdermal, subcutaneous, intrathecal, intramuscular, peritoneal, intraperitoneal, intracranial infusion or intravenous infusion.

[0243] The present invention also provides the following alternative embodiments:

[0244] 1. An antibody or antigen-binding fragment thereof that specifically binds to L1CAM, comprising a light chain variable region having light chain complementary determining regions LCDR1, LCDR2 and LCDR3 and / or a heavy chain variable region having heavy chain complementary determining regions HCDR1, HCDR2 and HCDR3, wherein the antibody or antigen-binding fragment thereof can specifically bind to the Ig1 or FN-4 domain of L1-CAM and does not have a CDR combination that is identical or equivalent to that of antibody L1-9.3 or UJ127.

[0245] 2. An antibody or antigen-binding fragment thereof according to embodiment 1, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having at least 80%, 90% or 100% identity with HCDR1, HCDR2 and HCDR3 of a heavy chain variable region sequence selected from SEQ ID NO: 43, 45, 47, 49 and 51, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having at least 80% identity with LCDR1, LCDR2 and LCDR3 of a light chain variable region sequence selected from SEQ ID NO: 44, 46, 48, 50 and 52.

[0246] 3. The antibody or antigen-binding fragment thereof according to embodiment 1 or 2, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region have a CDR combination selected from the following:

[0247] (1) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:43, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:44;

[0248] (2) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:45, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:46;

[0249] (3) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:47, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:48;

[0250] (4) HCDR1, HCDR2 and HCDR3 having at least 80% identity to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:49, and LCDR1, LCDR2 and LCDR3 having at least 80% identity to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:50; and

[0251] (5) HCDR1, HCDR2 and HCDR3 that are at least 80% identical to HCDR1, HCDR2 and HCDR3 of the heavy chain variable region sequence of SEQ ID NO:51, and LCDR1, LCDR2 and LCDR3 that are at least 80% identical to LCDR1, LCDR2 and LCDR3 of the light chain variable region sequence of SEQ ID NO:52.

[0252] 4. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 3, wherein:

[0253] (1) the HCDR1 has at least 80% identity with a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 13, SEQ ID NO: 19, SEQ ID NO: 25, SEQ ID NO: 31, SEQ ID NO: 37;

[0254] (2) the HCDR2 has at least 80% identity with a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 8, SEQ ID NO: 14, SEQ ID NO: 20, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 38;

[0255] (3) the HCDR3 has at least 80% identity with a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:15, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:33, SEQ ID NO:39;

[0256] (4) the LCDR1 has at least 80% identity with a sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:16, SEQ ID NO:22, SEQ ID NO:28, SEQ ID NO:34, SEQ ID NO:40;

[0257] (5) the LCDR2 has at least 80% identity with a sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:23, SEQ ID NO:29, SEQ ID NO:35, SEQ ID NO:41;

[0258] (6) The LCDR3 has at least 80% identity with a sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:42.

[0259] 5. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein:

[0260] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 1;

[0261] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 2;

[0262] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 3;

[0263] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 4;

[0264] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 5;

[0265] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 6.

[0266] 6. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein:

[0267] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 7;

[0268] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 8;

[0269] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 9;

[0270] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 10;

[0271] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 11;

[0272] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 12.

[0273] 7. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein: (1) the HCDR1 comprises the amino acid sequence as shown in SEQ ID NO: 13;

[0274] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 14;

[0275] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 15;

[0276] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 16;

[0277] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 17;

[0278] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 18.

[0279] 8. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein: (1) the HCDR1 comprises the amino acid sequence as shown in SEQ ID NO: 19;

[0280] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 20;

[0281] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 21;

[0282] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 22;

[0283] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 23;

[0284] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 24.

[0285] 9. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein: (1) the HCDR1 comprises the amino acid sequence as shown in SEQ ID NO: 25;

[0286] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 26;

[0287] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 27;

[0288] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 28;

[0289] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 29;

[0290] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 30.

[0291] 10. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein: (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 31;

[0292] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 32;

[0293] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 33;

[0294] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 34;

[0295] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 35;

[0296] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 36.

[0297] 11. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 4, wherein:

[0298] (1) the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 37;

[0299] (2) the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 38;

[0300] (3) the HCDR3 comprises the amino acid sequence shown in SEQ ID NO: 39;

[0301] (4) the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 40;

[0302] (5) the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 41;

[0303] (6) The LCDR3 comprises the amino acid sequence shown in SEQ ID NO: 42.

[0304] 12. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the heavy chain variable region comprises an amino acid sequence shown in SEQ ID NO: 43 or an amino acid sequence having at least 90%, at least 95% or 100% sequence identity with SEQ ID NO: 43.

[0305] 13. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 44 or an amino acid sequence having at least 90%, at least 95% or 100% sequence identity with SEQ ID NO: 44.

[0306] 14. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 45 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 45.

[0307] 15. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 46 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 46.

[0308] 16. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 47 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 47.

[0309] 17. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 48 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 48.

[0310] 18. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 9, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 49 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 49.

[0311] 19. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 50 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 50.

[0312] 20. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 51 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 51.

[0313] 21. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 11, wherein the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 52 or an amino acid sequence having at least 90%, at least 95% or higher sequence identity with SEQ ID NO: 52.

[0314] 22. The antibody or antigen-binding fragment thereof according to any one of embodiments 1-21, wherein the antibody is a humanized antibody.

[0315] 23. The antibody or antigen-binding fragment thereof according to embodiment 22, comprising a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 53 or SEQ ID NO: 54 or SEQ ID NO: 55 or SEQ ID NO: 56, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 57 or SEQ ID NO: 58 or SEQ ID NO: 59.

[0316] 24. The antibody or antigen-binding fragment thereof according to embodiment 23, comprising a heavy chain variable region and / or a light chain variable region, wherein:

[0317] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0318] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0319] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59; or

[0320] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0321] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0322] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59; or

[0323] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0324] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0325] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59; or

[0326] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57; or

[0327] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 58; or

[0328] 12) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 59.

[0329] 25. An antibody or antigen-binding fragment thereof according to embodiment 23, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 60 or SEQ ID NO: 61 or SEQ ID NO: 62 or SEQ ID NO: 63, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 64 or SEQ ID NO: 65 or SEQ ID NO: 66.

[0330] 26. The antibody or antigen-binding fragment thereof according to embodiment 25, comprising a heavy chain variable region and / or a light chain variable region, wherein:

[0331] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0332] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0333] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66; or

[0334] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0335] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0336] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66; or

[0337] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0338] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0339] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66; or

[0340] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64; or

[0341] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 65; or

[0342] 12) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 66.

[0343] 27. The antibody or antigen-binding fragment thereof according to embodiment 22, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 67 or SEQ ID NO: 68 or SEQ ID NO: 69 or SEQ ID NO: 70 or SEQ ID NO: 71 or SEQ ID NO: 72 or SEQ ID NO: 73 or SEQ ID NO: 74, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 75 or SEQ ID NO: 76 or SEQ ID NO: 77 or SEQ ID NO: 78 or SEQ ID NO: 79 or SEQ ID NO: 80 or SEQ ID NO: 81.

[0344] 28. The antibody or antigen-binding fragment thereof according to embodiment 27, comprising a heavy chain variable region and / or a light chain variable region, wherein:

[0345] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0346] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0347] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0348] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0349] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0350] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0351] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0352] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0353] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0354] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0355] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0356] 12) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0357] 13) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0358] 14) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0359] 15) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0360] 16) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0361] 17) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0362] 18) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0363] 19) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0364] 20) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0365] 21) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0366] 22) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0367] 23) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0368] 24) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0369] 25) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0370] 26) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0371] 27) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0372] 28) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 70, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0373] 29) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0374] 30) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0375] 31) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0376] 32) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0377] 33) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0378] 34) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0379] 35) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 71, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0380] 36) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0381] 37) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0382] 38) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0383] 39) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0384] 40) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0385] 41) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0386] 42) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0387] 43) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0388] 44) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0389] 45) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0390] 46) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0391] 47) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0392] 48) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0393] 49) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81; or

[0394] 50) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 75; or

[0395] 51) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 76; or

[0396] 52) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 77; or

[0397] 53) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 78; or

[0398] 54) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 79; or

[0399] 55) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 80; or

[0400] 56) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 81.

[0401] 29. The antibody or antigen-binding fragment thereof according to embodiment 22, wherein the heavy chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 82 or SEQ ID NO: 83 or SEQ ID NO: 84 or SEQ ID NO: 85 or SEQ ID NO: 86 or SEQ ID NO: 87 or SEQ ID NO: 88 or SEQ ID NO: 89 or SEQ ID NO: 90, and the light chain variable region comprises one of the amino acid sequences shown in SEQ ID NO: 91 or SEQ ID NO: 92 or SEQ ID NO: 93 or SEQ ID NO: 94 or SEQ ID NO: 95 or SEQ ID NO: 96.

[0402] 30. The antibody or antigen-binding fragment thereof according to embodiment 29, comprising a heavy chain variable region and / or a light chain variable region, wherein:

[0403] 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0404] 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0405] 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0406] 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0407] 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0408] 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0409] 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0410] 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0411] 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0412] 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0413] 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0414] 12) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0415] 13) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0416] 14) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0417] 15) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0418] 16) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0419] 17) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0420] 18) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0421] 19) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0422] 20) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0423] 21) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0424] 22) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0425] 23) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0426] 24) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0427] 25) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0428] 26) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0429] 27) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0430] 28) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0431] 29) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0432] 30) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0433] 31) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0434] 32) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0435] 33) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0436] 34) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0437] 35) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0438] 36) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0439] 37) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0440] 38) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0441] 39) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0442] 40) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0443] 41) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0444] 42) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0445] 43) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0446] 44) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0447] 45) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0448] 46) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0449] 47) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0450] 48) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96; or

[0451] 49) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 91; or

[0452] 50) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 92; or

[0453] 51) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 93; or

[0454] 52) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 94; or

[0455] 53) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 95; or

[0456] 54) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 96.

[0457] 31. The antibody or antigen-binding fragment thereof according to any one of embodiments 1-30, further comprising an Fc region, optionally comprising an Fc region of a human immunoglobulin (Ig), or optionally comprising an Fc region of a human IgG, and the Fc region is derived from human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM.

[0458] 32. An antibody or antigen-binding fragment thereof according to any one of embodiments 1-31, wherein the antigen-binding fragment is scFv, scFv-Fc, Fv, Fab, Fab', F(ab)2 or F(ab')2.

[0459] 33. The antibody or antigen-binding fragment thereof according to any one of embodiments 1-32, which is a monoclonal antibody, a bispecific antibody, a multispecific antibody, a recombinant antibody, a chimeric antibody, a bivalent antibody, an anti-idiotypic antibody or a fusion protein.

[0460] 34. An antibody-drug conjugate comprising the antibody or antigen-binding fragment of any one of embodiments 1-33, and an additional effector molecule.

[0461] 35. A nucleic acid molecule encoding the antibody or antigen-binding fragment of any one of embodiments 1-33.

[0462] 36. The nucleic acid molecule of embodiment 39, operably linked to a promoter.

[0463] 37. A recombinant expression vector, comprising the nucleic acid molecule of embodiment 39 or 40.

[0464] 38. A plasmid comprising the nucleic acid molecule of embodiment 39 or 40.

[0465] 39. A host cell comprising the nucleic acid molecule of embodiment 39 or 40, the recombinant expression vector of embodiment 41 or the plasmid of embodiment 42.

[0466] 40. A method for detecting L1CAM expression in a biological sample, comprising the following steps: contacting the biological sample with the antibody or antigen-binding fragment thereof according to any one of embodiments 1-33, and detecting the presence or absence of an immune complex, wherein the presence of the immune complex indicates the presence of L1CAM in the biological sample.

[0467] 41. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of embodiments 1-33, the antibody-drug conjugate according to any one of embodiments 34-38 as an active substance, and a pharmaceutically acceptable carrier.

[0468] 42. A kit comprising the antibody or antigen-binding fragment thereof of any one of embodiments 1-33, the antibody-drug conjugate of embodiment 34 and / or the pharmaceutical composition of embodiment 45.

[0469] 43. A method for treating cancer, comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof of any one of embodiments 1-33, the antibody-drug conjugate of embodiment 34, or the pharmaceutical composition of embodiment 45.

[0470] 44. The method according to embodiment 47, further comprising detecting the presence of L1-CAM in the cancer sample before administering the medicament.

[0471] In order to more thoroughly understand the invention, some definitions are listed below. The above definitions are intended to include grammatical equivalents.

[0472] "Antibody" herein means a protein consisting of one or more polypeptides encoded by all or part of a substantially recognized immunoglobulin gene. The recognized immunoglobulin genes, for example, in humans, include kappa (κ), lambda (λ) and heavy chain loci, which contain numerous variable region genes, and constant region genes mu (μ), delta (δ), gamma (γ), epsilon (ε), alpha (α) encoding IgM, IgD, IgG, IgE and IgA isotypes, respectively. Antibodies herein are meant to include full-length antibodies and antibody fragments, as well as natural antibodies from any organism, engineered antibodies, or antibodies recombinantly produced for experimental, therapeutic purposes or other purposes as further specified below. The term "antibody" includes antibody fragments, as known in the art, such as Fab, Fab', F(ab')2, Fv, scFv or other antigen-binding subsequences of antibodies, or antibody fragments produced by modifying intact antibodies or those synthesized de novo using recombinant DNA technology. The term "antibody" includes monoclonal and polyclonal antibodies. The antibody may be an antagonist, an agonist, a neutralizing antibody, or an inhibitory antibody, or a stimulatory antibody. The antibody of the present invention may be a non-human antibody, a chimeric antibody, a humanized antibody or a fully human antibody.

[0473] The term "monoclonal antibody" used herein refers to an antibody obtained from a substantially homogeneous antibody population, that is, the population comprising a single antibody is identical except for possible mutations (e.g., natural mutations) that may be present in very small amounts. Therefore, the term "monoclonal" indicates the nature of the antibody, i.e., it is not a mixture of unrelated (discrete) antibodies. Contrary to polyclonal antibody preparations that generally include different antibodies for different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed to a single determinant on an antigen. In addition to its specificity, the advantage of monoclonal antibody preparations is that they are generally not contaminated by other immunoglobulins. The term "monoclonal" should not be understood as requiring the antibody to be produced by any specific method. The term monoclonal antibody specifically includes fully human antibodies. (refCN201280067865-Anti-FGFR2 antibodies and their uses-approval authorization).

[0474] The term "complementarity determining region" (CDR, e.g., CDR1, CDR2, and CDR3) as used herein refers to the amino acid residues in the variable region of an antibody, the presence of which is essential for antigen binding. The "antigen binding region" of an antibody is typically present in one or more hypervariable regions of the antibody, such as the CDR regions of CDR1, CDR2, and CDR3. Each complementary determining region may comprise amino acid residues from a "complementarity determining region" as defined by Kabat, e.g., residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable region and 26-33 (H1), 51-57 (H2), and 96-105 (H3) in the heavy chain variable region (Kabat et al., Sequences of Proteins of Immulological Interest, 5th Ed. Public Health Service, National Institutes of Health, 1991).

[0475] As used herein, "antigen" means a compound, composition or substance that can stimulate antibody production or T cell response in an animal, including compositions injected or absorbed into an animal, which may be proteins, carbohydrates, lipids or other pathogens.

[0476] "Functional fragments" or "antigen-binding fragments" of antibodies / immunoglobulins are defined herein as fragments of antibodies / immunoglobulins that retain the antigen-binding region (e.g., the variable region of IgG). The "antigen-binding region" of an antibody is generally present in one or more hypervariable regions of the antibody, such as CDR1, -2 and / or -3 regions; however, the variable "framework" region can also play an important role in antigen binding, for example by providing a scaffold for the CDR. Preferably, the "antigen-binding region" includes at least amino acid residues 4-103 of the variable light chain (VL) and amino acid residues 5-109 of the variable heavy chain (VH), more preferably amino acid residues 3-107 of VL and amino acid residues 4-111 of VH, and particularly preferably complete VL and VH chains (amino acids 1-109 of VL and amino acids 1-113 of VH; numbering is performed according to WO97 / 08320). The preferred immunoglobulin class used in the present invention is IgG.

[0477] As used herein, "amino acid" means one of the 20 naturally occurring amino acids or any non-natural analogs, which may be located at a specifically defined position. As used herein, "protein" means at least two covalently linked amino acids, which include proteins, polypeptides, oligopeptides and peptides. Proteins may be composed of naturally occurring amino acids and peptide bonds, or of synthetic peptide mimetic structures, which peptide mimics are "analogs". Therefore, as used herein, "amino acid" or "peptide residue" means naturally occurring and synthetic amino acids. For example, for the purposes of the present invention, homophenylalanine, citrulline and norleucine are considered amino acids for the purposes of the present invention. "Amino acid" also includes imino acid residues such as proline and hydroxyproline. The side chain may be in the (R) or (S) configuration. In a preferred embodiment, the amino acid is present in the (S) or L-configuration. If non-naturally occurring side chains are used, non-amino acid substitutions may be used, for example to prevent or delay degradation in vivo.

[0478] As used herein, "identity" means the similarity between nucleotide or amino acid sequences, alternatively referred to as sequence identity. Sequence identity is typically measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar the two sequences are. When aligned using standard methods, homologues or variants will have a relatively high degree of sequence identity. Methods of sequence alignment for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith and Waterman, Adv Appl. Math., 2:482, 1981; Needlema and Wunsch, J. Mol. Biol. 48:443, 1970; Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85:2444, 1988; Higgins and Sharp, Gene 73:237-244, 1988; Higgins and Sharp, CABIOS 5:151-153, 1989; Corpet et al., Nucleic Acids Research 16:10881-10890, 1988; and Altschul et al., Nature Genet., 1994, 6:119-129.

[0479] The NCBI Basic Local Alignment Search Tool (BLAST™) (Altschul et al., J. Mol. Biol., 215:403-410, 1990) is available from several sources, including the National Center for Biotechnology Information (NCBI, Bethesda, Md.) and on the Internet for the programs blastp, blastn, blastx, tblastn and tblastx for sequence analysis.

[0480] " Nucleic acid " used herein means the polymer consisting of nucleotide units (ribonucleotides, deoxyribonucleotides, related naturally occurring structural variants and their synthesized non-naturally occurring analogs) by phosphodiester bonds. Therefore, the term includes nucleotide polymers, wherein nucleotides and the bonds therebetween include non-naturally occurring synthetic analogs, such as but not limited to thiophosphates, phosphoramidates, methyl phosphonates, chiral methyl phosphonates, 2'-O-methyl ribonucleotides, peptide nucleic acids (PNA) etc. For example, these polynucleotides can be synthesized using an automatic DNA synthesizer. The term " oligonucleotide " generally refers to short polynucleotides, generally not more than about 50 nucleotides. It should be understood that when the nucleotide sequence is represented by a DNA sequence (i.e. A, T, G, C), this also includes RNA sequences (i.e. A, U, G, C) in which "U" replaces "T".

[0481] Conventional notation is used herein to describe nucleotide sequences: the left-hand end of a single-stranded nucleotide sequence is the 5' end; the left-hand direction of a double-stranded nucleotide sequence is the 5' direction. The direction in which 5' to 3' nucleotides are added to the nascent RNA transcript is called the transcription direction. The DNA strand with the same sequence as the mRNA is called the coding strand.

[0482] As used herein, "coding" means the inherent properties of a specific nucleotide sequence in a polynucleotide, such as a gene, cDNA or mRNA, used as a template for synthesizing other polymers and macromolecules in a biological process with a determined nucleotide sequence, or a determined amino acid sequence and the resulting biological properties. Therefore, if the transcription and translation of the mRNA produced by the gene produces protein in a cell or other biological system, the gene encodes a protein. The coding strand (its nucleotide sequence is identical to the mRNA sequence and is generally provided in a sequence table) and the non-coding strand (used as a transcription template, gene or cDNA) can be referred to as a coded protein. Or other products of the gene or cDNA. Unless otherwise indicated, "nucleotide sequences encoding amino acid sequences" include all nucleotide sequences that are degenerate forms of each other and encode the same amino acid sequence. The nucleotide sequences encoding proteins and RNA may include introns.

[0483] As used herein, "plasmid" means a plasmid artificially constructed on the basis of a natural plasmid to adapt to laboratory operations. A nucleic acid molecule can be introduced into a host cell to produce a transformed host cell. A vector may include a nucleic acid sequence that allows it to replicate in a host cell, such as an origin of replication, and may also include one or more selectable marker genes and other genetic elements known in the art.

[0484] The "host cell" used herein is also called a recipient cell, and refers to a host cell that receives foreign genes during transformation and transduction (infection).

[0485] As used herein, "pharmaceutically acceptable carrier" means a conventional pharmaceutically acceptable carrier. Remington's Pharmaceutical Sciences, EW Martin, Mack Publishing Co., Easton, Pa., 15th edition (1975), describes compositions and formulations suitable for drug delivery of one or more therapeutic compounds or molecules (e.g., one or more antibodies), as well as additional agents.

[0486] As used herein, "diagnosing" a disease means determining a patient's condition and its progression after examination. "Preventing" a disease means inhibiting the full development of a disease. "Treatment" means therapeutic intervention to ameliorate the signs or symptoms of a disease or pathological condition after it has begun to develop.

[0487] "Administering" herein means selecting an appropriate route to introduce the substance into a subject. For example, if the selected route is intravenous, the composition is administered by introducing the substance into a vein of the subject.

[0488] Herein, "effective preventive / therapeutic dose" means a certain amount of a specific agent sufficient to achieve the desired effect in a subject treated with the agent. The exact dose will depend on the purpose of the treatment and can be determined by those skilled in the art using known techniques. The dosage range may be 0.01-100 mg / kg body weight or greater, such as 0.1, 1, 10 or 50 mg / kg body weight, preferably 1-10 mg / kg. As is known in the art, adjustments may be necessary for antibody or Fc fusion degradation, systemic or local drug delivery and new protease synthesis rates, as well as age, weight, general health status, sex, diet, administration time, drug interactions, and severity of the disease, and may be determined by those skilled in the art by conventional experimental methods. Such agents include monomeric Fc domain molecules described herein. In a non-limiting example, this may be the amount of HIV-specific monomeric Fc domains (or HIV-specific CH3 domain molecules) used to prevent, treat or improve HIV infection. Ideally, a therapeutically effective amount of an antibody is an amount sufficient to prevent, treat or ameliorate an infection or disease, such as caused by HIV infection in a subject without causing significant cytotoxic effects in the subject. The therapeutically effective amount of an agent used to prevent, ameliorate and / or treat a subject will depend on the subject being treated, the type and severity of the affliction, and the manner in which the therapeutic composition is administered.

[0489] "Cancer" herein is a solid tumor or hematogenous cancer. Solid tumors of the present invention are sarcomas or cancers, such as fibrosarcomas, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, or another sarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, lymphoid malignancies, pancreatic cancer, breast cancer, lung cancer, ovarian cancer, prostate cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, bile duct carcinoma, choriocarcinoma, Wilms' tumor, cervical cancer, testicular tumors, bladder cancer or central nervous system tumors (such as gliomas, astrocytomas, medulloblastomas, craniopharyngiomas, ependymomas, pineal glands, hemangioblastomas, acoustic neuromas, oligodendrogliomas, hemangiomas, melanomas, neuroblastomas or retinoblastomas). The blood-borne cancer described in the present invention is a leukemia, such as acute leukemia (such as acute lymphocytic leukemia, acute myeloid leukemia, acute myeloid leukemia and myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia); chronic leukemia (such as chronic myeloid (granulocytic) leukemia, chronic myeloid leukemia and chronic lymphocytic leukemia), polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma (indolent and advanced forms), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia or myelodysplasia.

[0490] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. Unless the context clearly indicates otherwise, the singular terms "a", "an" and "the" include plural indicators. It should also be understood that all base sizes or amino acid sizes given for nucleic acids or polypeptides, and all molecular weights or molecular weight values ​​are approximate and are provided for description. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. The term "comprising" means "including". All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, this specification (including term explanations) shall prevail. In addition, materials, methods and embodiments are illustrative only and not restrictive. BRIEF DESCRIPTION OF THE DRAWINGS

[0491] Figure 1 , Binding curve of mouse anti-L1CAM monoclonal antibody to HeLa cells.

[0492] Figure 2 , Binding curves of anti-L1CAM human-mouse chimeric antibody to HeLa cells and HeLa KO cells.

[0493] DETAILED DESCRIPTION OF THE EMBODIMENTS Definition

[0494] Unless otherwise defined, the technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art. In addition, the terms and laboratory procedures related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology used herein are terms and conventional procedures widely used in the corresponding fields. At the same time, in order to better understand the present invention, the definitions and explanations of the relevant terms are provided below.

[0495] The term "L1CAM" used in the present invention refers to any variant, isoform and species homolog of L1CAM. L1CAM is a transmembrane glycoprotein with a molecular weight between 200,000 and 220,000, also known as CD171. Structurally, the extracellular part of L1CAM is composed of 6 immunoglobulin-like domains and 5 type III fibronectin repeats starting from the amino terminus of the protein. In addition, L1CAM also contains a transmembrane domain and an intracellular domain. In certain embodiments, the L1CAM is human L1CAM. Exemplary sequences of human L1CAM proteins are disclosed in NCBI Ref Seq No. NP_001265045.1, NCBI RefSeq No. NP_000416.1, NCBI Ref Seq No. NP_076493.1 and NCBI Ref Seq No. NP_001137435.1. Exemplary predicted sequences of monkey L1CAM proteins are disclosed in NCBI Ref Seq No. XP_005594990.1, Ref Seq No. XP_005594993.1, Ref Seq No. XP_005594994.1, Ref Seq No. XP_005594995.1, and Ref Seq No. XP_005594996.1.

[0496] The term "antibody" as used in the present invention refers to any immunoglobulin that can bind to a specific antigen. A natural complete antibody contains two heavy (H) chains and two light (L) chains. Each heavy chain of a mammal is composed of a variable region (VH) and the first, second, third and fourth (optionally) constant regions (CH1, CH2, CH3, CH4, respectively); the light chains of mammals can be divided into λ or κ, and each light chain is composed of a variable region (VL) and a constant region. The variable regions of the light and heavy chains determine the binding of the antigen. The variable region of each chain usually contains three hypervariable regions, called complementarity determining regions (CDRs) (the light chain CDRs include LCDR1, LCDR2, LCDR3, and the heavy chain CDRs include HCDR1, HCDR2, HCDR3). The CDR boundaries of the antibodies and antigen-binding fragments disclosed in the present invention can be named or identified by Kabat nomenclature.

[0497] The term "monoclonal antibody" as used herein refers to a population of identical antibodies, meaning that each individual antibody molecule in the monoclonal antibody population is identical to the other antibody molecules. This property is in contrast to the properties of a polyclonal population of antibodies, which comprises antibodies with a variety of different sequences.

[0498] The term "bispecific antibody" used in the present invention refers to an artificial antibody that has fragments derived from two different monoclonal antibodies and can bind to two different epitopes. The two epitopes can exist on the same antigen, or they can exist on two different antigens.

[0499] The term "multispecific antibody" used in the present invention refers to an artificial antibody that has fragments derived from multiple different monoclonal antibodies and can bind to multiple different epitopes. The multiple epitopes can exist on the same antigen, or they can exist on multiple different antigens.

[0500] The term "recombinant antibody" used in the present invention refers to an antibody expressed in an engineered cell line or an engineered bacterial strain through recombinant genetic engineering technology.

[0501] The term "chimeric antibody" used in the present invention refers to an antibody molecule in which the antibody constant region or a portion thereof is replaced such that the antigen binding site (variable region, CDR or a portion thereof) is linked to a constant region of a different species.

[0502] The term "amino acid" as used herein refers to an organic compound containing amino (-NH2) and carboxyl (-COOH) functional groups and side chains unique to each amino acid. The amino acid names are also represented in the present invention by standard single-letter or three-letter codes recognized in the art.

[0503] As used in the present invention, "treatment" or "therapy" for a disease, disorder or condition includes preventing or alleviating a disease, disorder or condition, reducing the rate of occurrence or development of a disease, disorder or condition, reducing the risk of developing a disease, disorder or condition, preventing or delaying the development of symptoms associated with a disease, disorder or condition, reducing or stopping symptoms associated with a disease, disorder or condition, producing complete or partial reversal of a disease, disorder or condition, curing a disease, disorder or condition, or a combination of the above.

[0504] As used herein, the term "diagnosis" refers to the identification of a pathological state, disease or condition, such as the identification of a cancer-related disease, or to the identification of a subject who may benefit from a particular treatment regimen. Example

[0505] The present invention will be further described below by way of examples, but the present invention is not limited to the described examples.

[0506] Example 1: Production of monoclonal antibodies against L1CAM

[0507] In order to produce antibodies against L1CAM, Balb / c mice were immunized with recombinantly expressed human L1CAM antigen fragments. During the immunization operation, the mice were bled regularly to obtain serum samples and monitor the immune response. Mice with sufficient titers of anti-L1CAM antibodies were used for fusion experiments. Splenocytes and / or lymph node cells from immunized mice were isolated and fused with mouse myeloma cell lines. Hybridomas that can produce L1CAM-specific antibodies were screened by using HeLa cells that highly express human L1CAM and HEK293 cells that highly express monkey L1CAM by ELISA analysis with human L1CAM extracellular domain recombinant protein, and subcloned to obtain stable hybridoma clones. After subcloning, antibody sequencing was performed on monoclonal hybridoma cells for further analysis and testing.

[0508] In the present invention, the mouse anti-L1CAM monoclonal antibody is represented by the "hybridoma clone number", for example, the mouse anti-L1CAM monoclonal antibody from the hybridoma clone 25F8H8 is represented by "25F8H8".

[0509] Example 2: Production of chimeric antibodies against L1CAM

[0510] According to the results of monoclonal hybridoma cell antibody sequencing, human-mouse chimeric antibodies with CH1, CH2 and CH3 of human IgG1 as constant regions and VH and VL of mouse anti-L1CAM monoclonal antibody as variable regions were designed, recombinantly expressed in mammalian cells, and purified for further analysis and testing. In the present invention, the chimeric antibody number is represented as "ch+hybridoma clone number", for example, the chimeric antibody from hybridoma clone 25F8H8 is represented as "ch25F8H8".

[0511] Example 3: Binding of mouse anti-L1CAM monoclonal antibody to HeLa cells

[0512] The HeLa cells used in this example were used to determine the binding of mouse anti-L1CAM monoclonal antibodies to endogenously expressed human L1CAM on the cell surface. HeLa cells were cultured in DMEM complete medium (purchased from Corning) containing 10% FBS (purchased from Gibco) and a 100U / mL penicillin-streptomycin mixture (Thermofisher, 15140122). When the cell coverage in the culture container reached 80-90%, the HeLa cells were digested from the culture container using TrypLE (purchased from Thermofisher) and plated at 4x10 6 Cells were seeded at a density of 1:1 cells / ml in a 96-well plate (purchased from AXYGEN), with 100,000 cells per well (25 μL DMEM complete medium), and then 25 μL of diluted hybridoma supernatant containing antibodies was added to each well, gently mixed, and the 96-well plate was incubated at 4°C for 1 hour. Then 150 μL of PBS buffer containing 2% FBS (purchased from Corning) was added to each well, centrifuged at 1400 rpm for 4 minutes, and the supernatant was removed and washed again. Then 50 μL of 1:800 diluted AlexaFlour 647 labeled goat anti-mouse fluorescent secondary antibody (purchased from Jacksonimmuno) was added to each well, and the 96-well plate was incubated at 4°C for 0.5 hours. The cells were washed twice with PBS solution containing 2% FBS, and then the cells were resuspended in 120 μL PBS, and the fluorescence signal on the cell surface was detected using a flow cytometer (BD FACSCelesta) to obtain the median fluorescence intensity (MFI). The concentration of mouse IgG in the supernatant was determined by anti-mouse ELISA analysis of the hybridoma supernatant containing antibodies. Finally, a dose-response curve was generated and the half effective concentration (EC) was calculated by nonlinear regression using GraphPad software. 50 ).

[0513] Table 1. Binding of mouse anti-L1CAM monoclonal antibodies to HeLa cells

[0514] Antibody ID <![CDATA[EC 50 (nM)]]> 25F8H8 0.1927 26A12C1 0.2414 46F1D5 0.1440 49E10H1 0.2333 103E9B3 0.4356

[0515] Example 4: Binding of anti-L1CAM human-mouse chimeric antibody to human L1CAM endogenously expressed on HeLa cells

[0516] The HeLa cells and human L1CAM knockout HeLa cells (purchased from Abcam) used in this example were used to determine the binding of mouse anti-L1CAM monoclonal antibodies to endogenously expressed human L1CAM on the cell surface. Human L1CAM knockout HeLa cells are referred to as HeLa KO cells in the present invention.

[0517] Hela cells were cultured in DMEM complete medium (purchased from Corning) containing 10% FBS (purchased from Gibco) and 100 U / mL penicillin-streptomycin mixture (Thermofisher, 15140122). When the cell coverage rate in the culture vessel reached 80-90%, Hela and Hela KO cells were digested from the culture vessel using TrypLE (purchased from Thermofisher) and plated at 4×10 6 Cells / ml were seeded in a 96-well plate (purchased from AXYGEN), with 100,000 cells per well (25 μL DMEM complete medium), and then the human-mouse chimeric antibody was serially diluted with DMEM complete medium, and 25 μL of chimeric antibody dilution was added to each well, gently mixed, and the 96-well plate was incubated at 4°C for 1 hour. Then 150 μL of PBS buffer containing 2% FBS (purchased from Corning) was added to each well, centrifuged at 1400 rpm for 4 minutes, and the supernatant was removed and washed again. Then 50 μL of 1:800 diluted Alexa Flour 647 labeled goat anti-human fluorescent secondary antibody (purchased from Jacksonimmuno) was added to each well, and the 96-well plate was incubated at 4°C for 0.5 hours. The cells were washed twice with PBS solution containing 2% FBS, and then resuspended in 120 μL PBS. The fluorescence signal on the cell surface was detected using a flow cytometer (BD FACSCelesta) to obtain the median fluorescence intensity (MFI). The dose-response curve was then generated, and the half effective concentration (EC) was calculated by nonlinear regression using GraphPad software. 50 ). HeLa cells highly express L1CAM endogenously. Since the L1CAM gene is knocked out in HeLa KO cells, the expression level of L1CAM is significantly reduced compared with wild-type HeLa. Therefore, the binding ability of antibodies to L1CAM on the surface of HeLa cells can be determined by flow cytometry, and the specificity of antibodies can be evaluated by detecting the fluorescent signal on the surface of HeLa KO cells.

[0518] Table 2. Binding of anti-L1CAM chimeric antibodies to HeLa cells and HeLa KO cells

[0519]

[0520] Note: N / A means no data available.

[0521] Example 5: Production of humanized antibodies against L1CAM

[0522] Mouse anti-L1CAM monoclonal antibodies 46F1D5, 49E10H1 and 103E9B3 were selected as humanized clones. The antibody sequences were aligned with human germline sequences to determine the best fitting model. Based on the homology with the original mouse antibody sequence, the best matching human germline sequence was selected as the template for humanization. The CDRs from the mouse antibody sequence were then grafted onto the template together with residues that maintained the upper core and central core structures of the antibody. Optimized mutations were introduced into the framework region to generate variants of the humanized heavy chain variable region and variants of the humanized light chain variable region, which were mixed and matched to provide multiple humanized antibody clones. Recombinant expression was performed in mammalian cells and purified for further analysis and testing.

[0523] Example 6 Determination of humanized L1CAM antibody binding on the surface of Hela cells

[0524] The experimental principle is summarized as follows: L1CAM protein is highly expressed on the surface of Hela cells. After incubating the humanized L1CAM antibody and cells, the antibody's binding ability to LICAM on the surface of Hela cells can be determined by flow cytometry.

[0525] The experimental method is outlined as follows:

[0526] The humanized antibody against L1CAM (Human IgG1-Kappa) was diluted 4-fold in series using complete culture medium, with a total of 7 concentration gradient points.

[0527] Hela cells were cultured in DMEM complete medium (Corning, 10-013-CVR) containing 10% FBS (Gibco, 10099-141) and 100U / mL penicillin-streptomycin mixture (Thermofisher, 15140122). When the cell coverage in the culture vessel reached 80-90%, Hela cells were digested from the culture vessel using TripLE (Thermofisher, 12604-013) and seeded in a 96-well plate (AXYGEN, P-96-450V-C) at a density of 2.2*106 cells / ml, with 100,000 cells per well (45μL complete medium). Then, 5μL of antibody dilution was added to each well, gently mixed, and the 96-well plate was incubated at 4°C for 1 hour. Then add 150 μL of PBS (Corning, 21-040-CVC) solution containing 2% FBS to each well, centrifuge at 1400 rpm for 4 minutes, remove the supernatant, and repeat the washing once. Then add 50 μL of 1:800 diluted Alexa Flour 647 labeled goat anti-human fluorescent secondary antibody (Jacksonimmuno, 109-605-088) to each well, and incubate the 96-well plate at 4°C for 0.5 hours. Wash the cells twice with PBS solution containing 2% FBS, then resuspend the cells in 120 μL of PBS, and use a flow cytometer (BD FACS Celesta) to detect the fluorescent signal on the cell surface to obtain the median fluorescence intensity (MFI). Then generate a dose-response curve, and calculate the half effective concentration (EC) by nonlinear regression using GraphPad software. 50 ).

[0528] Table 3 Binding ability of humanized 26A12C1 antibody to Hela cells

[0529]

[0530]

[0531] Table 4 Binding ability of humanized 49E10H1 antibody to Hela cells

[0532] Humanized Antibody No. <![CDATA[EC 50 (nM)]]> hz49E10H1H4L5 2.15 hz49E10H1H4L6 0.8533 hz49E10H1H4L7 0.6764 hz49E10H1H5L5 0.6326 hz49E10H1H5L6 0.4042 hz49E10H1H5L7 0.3767 hz49E10H1H6L5 24.33 hz49E10H1H6L6 1.03 hz49E10H1H6L7 0.9962 hz49E10H1H7L5 10.39 hz49E10H1H7L6 1.051 hz49E10H1H7L7 0.8516 hz49E10H1H8L5 9.274 hz49E10H1H8L6 1.06 hz49E10H1H8L7 0.8204

[0533] Table 5 Binding ability of humanized 103E9B3 antibody to Hela cells

[0534] Humanized Antibody No. <![CDATA[EC 50 (nM)]]> ch103E9B3 1.21 hz103E9B3H5L4 6.845 hz103E9B3H5L5 1.682 hz103E9B3H5L6 1.926 hz103E9B3H6L4 6.595 hz103E9B3H6L5 1.483 hz103E9B3H6L6 2.184 hz103E9B3H7L4 5.323 hz103E9B3H7L5 2.133 hz103E9B3H7L6 2.034 hz103E9B3H8L4 5.795 hz103E9B3H8L5 1.54 hz103E9B3H8L6 1.997 hz103E9B3H9L4 5.564 hz103E9B3H9L5 1.964 hz103E9B3H9L6 2.065

[0535] In the present invention, the humanized antibody number is expressed as "hz+hybridoma clone number+heavy chain number+light chain number", for example, the humanized antibody "hz46F1D5H1L2" represents a humanized antibody derived from the mouse anti-L1CAM monoclonal antibody 46F1D5, and the variable region heavy chain is VH1 (i.e., SEQ ID NO: 60), and the variable region light chain is VL2 (i.e., SEQ ID NO: 65).

[0536] In the present invention, the human-mouse chimeric antibody number is expressed as "ch+hybridoma clone number", for example, the human-mouse chimeric antibody "ch46F1D5" represents a human-mouse chimeric antibody derived from the mouse anti-L1CAM monoclonal antibody 46F1D5, and the variable region heavy chain is the heavy chain variable region of the mouse monoclonal antibody 46F1D5, the variable region light chain is the light chain variable region of the mouse monoclonal antibody 46F1D5, and the constant region is the human IgG1 constant region.

[0537] Antibody sequence information:

[0538] Table 6 Amino acid sequences of the complementary determining regions of mouse monoclonal antibodies

[0539]

[0540]

[0541] Table 7 Mouse monoclonal antibody amino acid sequence

[0542]

[0543]

[0544] Table 8 Amino acid sequences of humanized monoclonal antibody variable regions

[0545]

[0546]

[0547]

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554]

[0555]

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565]

[0566]

[0567]

[0568]

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to L1CAM, comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region and the heavy chain variable region have: (a) HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 19, SEQ ID NO: 20, and SEQ ID NO: 21, respectively, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, respectively; (b) HCDR1, HCDR2, HCDR3 as shown in SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, respectively, and LCDR1, LCDR2, LCDR3 as shown in SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, respectively; (c) HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively; (d) HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9, respectively, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, respectively; (e) HCDR1, HCDR2, HCDR3 as shown in SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, respectively, and LCDR1, LCDR2, LCDR3 as shown in SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, respectively; or (f) HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:37, SEQ ID NO:38, and SEQ ID NO:39, respectively, and LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO:40, SEQ ID NO:41, and SEQ ID NO:42, respectively.

2. The antibody or antigen-binding fragment thereof according to claim 1, comprising a heavy chain variable region and a light chain variable region, wherein: 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 43, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 44; 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 45, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 46; 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 47, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 48; 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 49, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 50; or 5) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 51, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:

52.

3. The antibody or antigen-binding fragment thereof according to claim 1, comprising a heavy chain variable region and a light chain variable region, wherein: 1) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 53, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 57-59; 2) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 57-59; 3) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 57-59; 4) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 56, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 57-59; 5) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 64-66; 6) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 61, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 64-66; 7) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 62, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 64-66; 8) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 64-66; 9) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 67, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 75-81; 10) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 68, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 75-81; 11) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 69, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 75-81; 12) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 72, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 75-81; 13) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 73, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 75-81; 14) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 74, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 75-81; 15) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 82, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 16) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 83, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 17) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 84, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 18) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 85, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 19) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 86, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 20) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 87, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 21) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 88, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; 22) the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 89, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96; or 23) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 90, and the light chain variable region comprises the amino acid sequence shown in any one of SEQ ID NOs: 91-96.

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, further comprising an Fc region, wherein the Fc region comprises an Fc region of a human immunoglobulin (Ig). The antibody or antigen-binding fragment thereof according to claim 4 , wherein the Fc region is the Fc region of human IgG. The antibody or antigen-binding fragment thereof according to claim 4 , wherein the Fc region is derived from human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM.

7. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antigen-binding fragment is scFv, scFv-Fc, Fv, Fab, Fab', F(ab)2 or F(ab')2. The antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody is a monoclonal antibody.

9. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody is a chimeric antibody.

10. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody is a humanized antibody.

11. A nucleic acid molecule encoding the antibody or antigen-binding fragment of any one of claims 1-10.

12. The nucleic acid molecule of claim 11, operably linked to a promoter.

13. A recombinant expression vector, comprising the nucleic acid molecule according to claim 11 or 12.

14. A plasmid comprising the nucleic acid molecule according to claim 11 or 12.

15. A host cell comprising the nucleic acid molecule according to claim 11 or 12, the recombinant expression vector according to claim 13 or the plasmid according to claim 14.

16. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10 in preparing a preparation for detecting L1CAM expression in a biological sample.

17. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, and a pharmaceutically acceptable carrier.

18. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10 in the preparation of an agent for diagnosing L1CAM-positive cancer or a medicament for treating L1CAM-positive cancer.

19. A kit comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10.

Citation Information

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