Improved b7-h3 antibody and use thereof

Abstract

Provided are an improved B7-H3 antibody and the use thereof. Specifically provided are an antibody that specifically binds to B7-H3 or an antigen-binding fragment thereof, and the use thereof.

Description

Improved B7-H3 antibody and its application Technical Field

[0001] This disclosure relates to the biomedical field, and in particular to the use of antibodies targeting B7-H3 or antigen-binding fragments thereof, and their applications. Background Technology

[0002] B7-H3 (CD276) is a type I transmembrane protein located on human chromosome 15, with a molecular weight of 45kD–66kD. It is one of the co-stimulatory molecules in the B7 family and shares 20%–27% amino acid sequence homology with other family members. Structurally, B7-H3 has extracellular IgV / IgC tandem repeat sequences, a transmembrane region, and an intracellular domain (similar to PD-L1). Based on the number of extracellular tandem repeat units, two forms of B7-H3 have been identified: 2Ig-B7-H3 and 4Ig-B7-H3 (with one more IgV / C repeat unit), with 4Ig-B7-H3 considered more common.

[0003] B7-H3 mRNA is widely distributed, constitutively expressed in non-immune resting fibroblasts, endothelial cells, osteoblasts, and amniotic fluid stem cells, and inducibly expressed in activated T cells, NK cells, dendritic cells, and macrophages. In normal tissues, B7-H3 is negative in many tissues, but low to moderate expression has been detected in tissues such as the pancreas, liver, colon, stomach, placenta, skin, and adrenal glands.

[0004] Studies have shown that B7-H3 mRNA is overexpressed in various tumors, including breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, and thyroid cancer. Multiple studies have demonstrated that B7-H3 plays a crucial role in tumor progression, including promoting tumor proliferation / migration, mediating epithelial-mesenchymal transition (EMT) in tumor cells, and influencing tumor cell metabolism. B7-H3 expression is regulated by oncogenes, and upregulation of B7-H3 promotes tumor growth and metastasis through multiple signaling pathways.

[0005] Antibodies targeting B7-H3 can be used to treat or prevent diseases associated with B7-H3. However, antibody aggregation is a common problem. In vivo, protein folding is primarily driven by the embedding of hydrophobic residues, and the exposure of these residues can lead to non-natural self-binding, misfolding, and ultimately aggregation. The formation of such misfolded aggregates is triggered by a variety of factors, including hydrophobic regions and charge distribution. Antibody aggregation may be triggered by the partial unfolding and exposure of antibody domains, leading to monomer-monomer association, followed by the formation of nuclear seeds and subsequent growth. Such aggregation can reduce antibody activity and may trigger an immune response.

[0006] The sensitivity of IgG antibody aggregation is largely regulated by the CDR region. The CDR region of an antibody is the core part of antigen binding. Generally, the amino acid composition contains a high frequency of aromatic and hydrophobic amino acids (e.g., Tyr, Phe, Leu, and Ile), as well as amino acids (e.g., Ser, Thr, Asn, and Gln) as hydrogen bond donors.

[0007] Since the amino acids in the CDR region are primarily responsible for antigen binding, designing CDR region modifications to resist antibody aggregation without compromising the original antigen affinity activity is a major challenge in antibody aggregation modification. Summary of the Invention

[0008] This disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to B7-H3 and exhibit improved thermal stability, capable of resisting aggregation. Based on existing B7-H3 antibodies (WO2024114525A1, the entire contents of which are incorporated herein by reference), this disclosure, through analysis and screening, reveals that amino acid substitution at specific sites on the antibody (particularly the CDR region) enhances its anti-aggregation properties without affecting antigen affinity, thereby avoiding adverse effects such as reduced antibody activity and triggering immune responses. Simultaneously, it increases the dissociation rate of the antibody from the antigen under acidic pH conditions, thus contributing to a longer antibody half-life and improved stability and bioavailability.

[0009] According to one aspect of this disclosure, an antibody or antigen-binding fragment thereof specifically binds to B7-H3 is provided, said antibody or antigen-binding fragment comprising: an amino acid residue of asparagine (Asn, N) or tyrosine (Tyr, Y) at position 34 of the heavy chain variable region; an amino acid residue of histidine (His, H) at position 57 of the heavy chain variable region; and / or an amino acid residue of D at position 93 of the light chain variable region.

[0010] In some embodiments, the amino acid positions in the heavy chain variable region are defined relative to the amino acid sequence shown in SEQ ID NO:1. In some embodiments, the amino acid positions in the light chain variable region are defined relative to the amino acid sequence shown in SEQ ID NO:7.

[0011] In some embodiments, the antibody or its antigen-binding fragment may include an amino acid residue with N at position 34 and an amino acid residue with F at position 57 in the variable region of the heavy chain.

[0012] In some embodiments, the antibody or its antigen-binding fragment may include an amino acid residue with N at position 34 and an amino acid residue with H at position 57 in the variable region of the heavy chain.

[0013] In some embodiments, the antibody or its antigen-binding fragment may include an amino acid residue with position Y at position 34 and an amino acid residue with position F at position 57 in the variable region of the heavy chain.

[0014] In some embodiments, the antibody or its antigen-binding fragment may include an amino acid residue with position Y at position 34 and an amino acid residue with position H at position 57 in the variable region of the heavy chain.

[0015] In some embodiments, the antibody or its antigen-binding fragment may include an amino acid residue with a W at position 34 and an amino acid residue with an H at position 57 in the variable region of the heavy chain.

[0016] In some embodiments, the antibody or its antigen-binding fragment may further include an amino acid residue with a D at position 93 in the variable region of the light chain.

[0017] In some embodiments, the antibody or its antigen-binding fragment may include:

[0018] (1) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0019] (2) The heavy chain variable region (VH) comprises the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain variable region (VL) comprises the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0020] (3) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:3, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0021] (4) Heavy chain variable region (VH), comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO:3; and / or, light chain variable region (VL), comprising an amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0022] (5) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0023] (6) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0024] (7) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0025] (8) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0026] (9) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0027] (10) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or

[0028] (11) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:1, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0029] In some embodiments, the antibody or its antigen-binding fragment may also include a constant region of an immunoglobulin derived from a human or mouse.

[0030] In some embodiments, the antibody or its antigen-binding fragment may include: (1) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0031] (2) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0032] (3) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:39, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0033] (4) A heavy chain comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO:39; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0034] (5) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0035] (6) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0036] (7) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0037] (8) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0038] (9) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0039] (10) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0040] (11) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:37, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0041] (12) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0042] (13) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0043] (14) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:47, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0044] (15) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:48, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0045] (16) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0046] (17) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0047] (18) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or

[0048] (19) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0049] According to another aspect of this disclosure, antibodies or antigen-binding fragments thereof that specifically bind to B7-H3 are provided, including:

[0050] (1) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:2; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:7.

[0051] (2) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:2; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8.

[0052] (3) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:3; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:7.

[0053] (4) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:3; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8.

[0054] (5) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:4; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:7.

[0055] (6) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:4; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8.

[0056] (7) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:5; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:7.

[0057] (8) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:5; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8.

[0058] (9) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:6; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:7.

[0059] (10) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:6; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8.

[0060] (11) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:1; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8; or

[0061] (12) Having one or more amino acid substitutions, deletions or additions compared to CDR-H1, CDR-H2 and CDR-H3 as shown in any of (1) to (11); and / or having one or more amino acid substitutions, deletions or additions compared to CDR-L1, CDR-L2 and CDR-L3 as shown in any of (1) to (11).

[0062] In some implementations, the CDR is defined according to a numbering system of IMGT, Kabat, Chothia, AbM, or combinations thereof.

[0063] In some embodiments, the CDR is defined according to the IMGT numbering system. In such embodiments, the antibody or its antigen-binding fragment comprises:

[0064] (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:15, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0065] (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:15, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0066] (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0067] (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0068] (5) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0069] (6) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0070] (7) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0071] (8) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0072] (9) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0073] (10) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0074] (11) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:15, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or

[0075] (12) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-H1, CDR-H2 and CDR-H3 as shown in (1) to (11); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-L1, CDR-L2 and CDR-L3 as shown in (1) to (11).

[0076] In some embodiments, the CDR is defined according to the Chothia numbering system. In such embodiments, the antibody or its antigen-binding fragment comprises:

[0077] (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:23, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0078] (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:23, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0079] (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:22, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0080] (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:22, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or

[0081] (5) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-H1, CDR-H2 and CDR-H3 as shown in (1) to (4); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-L1, CDR-L2 and CDR-L3 as shown in (1) to (4).

[0082] In some embodiments, the CDR is defined according to the Kabat numbering system. In such embodiments, the antibody or its antigen-binding fragment comprises:

[0083] (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:25, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0084] (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:25, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0085] (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0086] (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0087] (5) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0088] (6) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0089] (7) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0090] (8) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0091] (9) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0092] (10) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0093] (11) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:25, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or

[0094] (12) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-H1, CDR-H2 and CDR-H3 as shown in (1) to (11); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-L1, CDR-L2 and CDR-L3 as shown in (1) to (11).

[0095] In some embodiments, the CDR is defined according to the AbM numbering system. In such embodiments, the antibody or its antigen-binding fragment comprises:

[0096] (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:30, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0097] (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:30, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0098] (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0099] (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0100] (5) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0101] (6) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0102] (7) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0103] (8) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0104] (9) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:11.

[0105] (10) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0106] (11) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:30, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12.

[0107] (12) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-H1, CDR-H2 and CDR-H3 as shown in (1) to (11); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-L1, CDR-L2 and CDR-L3 as shown in (1) to (11).

[0108] Those skilled in the art should understand that the above-mentioned amino acid substitutions are conservative substitutions.

[0109] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0110] (1) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0111] (2) The heavy chain variable region (VH) comprises the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, the light chain variable region (VL) comprises the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0112] (3) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:3, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0113] (4) Heavy chain variable region (VH), comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO:3; and / or, light chain variable region (VL), comprising an amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0114] (5) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0115] (6) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0116] (7) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0117] (8) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0118] (9) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0119] (10) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or

[0120] (11) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:1, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0121] In some embodiments, the antibody or antigen-binding fragments disclosed herein include antibodies or antigen-binding fragments that specifically bind to antigens and may include post-translational modifications thereof (e.g., C-terminal lysine cleavage in the heavy chain, N-terminal glutamine or glutamate conversion to pyroglutamic acid or pyroglutamate salt in the heavy or light chain), which may occur during recombinant expression in host cells (e.g., CHO cells) or during purification / storage.

[0122] In some embodiments of the antibody or antigen-binding fragment disclosed herein, the heavy chain constant domain may contain a C-terminal lysine residue or lack a C-terminal lysine residue or a C-terminal glycine-lysine dipeptide. In some embodiments of the antibody or antigen-binding fragment thereof, the N-terminal amino acid of the antibody or antigen-binding fragment thereof may be cyclized to pyroglutamic acid. In some embodiments of the antibody or antigen-binding fragment thereof, the N-terminal amino acid of the antibody or antigen-binding fragment thereof may be cyclized to pyroglutamic acid salt.

[0123] As is known to those skilled in the art, pyroglutamic acid is the conjugate acid of pyroglutamate and is in equilibrium with pyroglutamate in solution.

[0124] In some embodiments, the heavy and / or light chains of the antibody or antigen-binding fragment may include a C-terminal lysine, a lack of a C-terminal lysine, or a lack of a C-terminal glycine-lysine.

[0125] In some embodiments, the heavy and / or light chains of the antibody or antigen-binding fragment may include N-terminal glutamine or glutamate, N-terminal amino acid cyclized to pyroglutamate, or N-terminal amino acid cyclized to pyroglutamate. In some embodiments, compositions comprising the antibody or antigen-binding fragments disclosed herein are provided, wherein the various antibody or antigen-binding fragments may independently comprise C-terminal lysine, lack C-terminal lysine, lack C-terminal glycine-lysine, and / or comprise N-terminal glutamine or glutamate, N-terminal amino acid cyclized to pyroglutamate, or N-terminal amino acid cyclized to pyroglutamate.

[0126] In some embodiments, the N-terminal glutamine of the heavy chain variable region having a sequence or a variant thereof as shown in any one of SEQ ID NO:1, 2, 3, 4, 5 or 6 undergoes cyclization to form pyroglutamic acid or pyroglutamate, and / or, the C-terminus of the heavy chain constant region having a sequence or a variant thereof as shown in SEQ ID NO:35 or 45 lacks lysine.

[0127] In some embodiments, the antibody or its antigen-binding fragment may further include at least a portion of a constant region derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, or goats. In some embodiments, the antibody or its antigen-binding fragment may include at least a portion of a constant region derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans, preferably a constant region derived from human immunoglobulins.

[0128] In some embodiments, at least a portion of the heavy chain constant region of the antibody or its antigen-binding fragment may include a heavy chain constant region derived from IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4 or IgM immunoglobulin.

[0129] In some embodiments, the antibody or its antigen-binding fragment may include a wild-type heavy chain constant region or a mutated or chemically modified heavy chain constant region. The mutated or chemically modified heavy chain constant region may have altered effector functions (e.g., reduced ADCC activity) compared to the wild-type heavy chain constant region.

[0130] In some embodiments, the antibody or its antigen-binding fragment may include a heavy chain constant region derived from human IgG1, or a variant thereof containing mutations or chemical modifications.

[0131] In some embodiments, the heavy chain constant region of the antibody or its antigen-binding fragment may include the amino acid sequence shown in SEQ ID NO:35 or 45, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or having at most 20, at most 19, at most 18, at most 17, at most 16, at most 15, at most 14, at most 13, at most 12, at most 11, at most 10, at most 9, at most 8, at most 7, at most 6, at most 5, at most 4, at most 3, at most 2, or at most 1 conserved amino acid substitutions compared to it.

[0132] In some embodiments, at least a portion of the light chain constant region of the antibody or its antigen-binding fragment may include light chain constant regions derived from human κ and λ immunoglobulins, or variants thereof containing mutations or chemical modifications.

[0133] In some embodiments, the light chain constant region of the antibody or its antigen-binding fragment may include the amino acid sequence shown in SEQ ID NO:36, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or having at most 20, at most 19, at most 18, at most 17, at most 16, at most 15, at most 14, at most 13, at most 12, at most 11, at most 10, at most 9, at most 8, at most 7, at most 6, at most 5, at most 4, at most 3, at most 2, or at most 1 conserved amino acid substitutions with it.

[0134] In some embodiments, the antibody or its antigen-binding fragment may be a murine antibody, a chimeric antibody, a humanized antibody, or a fully human antibody.

[0135] In some embodiments, the antibody or its antigen-binding fragment may include:

[0136] (1) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or, a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0137] (2) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0138] (3) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:39, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0139] (4) A heavy chain comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO:39; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0140] (5) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0141] (6) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0142] (7) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0143] (8) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0144] (9) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0145] (10) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or

[0146] (11) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:37, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0147] (12) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0148] (13) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0149] (14) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:47, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0150] (15) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:48, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0151] (16) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0152] (17) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0153] (18) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it, or

[0154] (19) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with it.

[0155] In some embodiments, the N-terminal glutamine of a heavy chain having a sequence or a variant thereof as shown in any one of SEQ ID NO:37, 38, 39, 40, 41, 42 or 46-50 undergoes cyclization to form pyroglutamic acid or pyroglutamate, and / or, the C-terminus lacks lysine.

[0156] In some embodiments, the antigen-binding fragment of the antibody may include, but is not limited to, Fab fragment, Fab' fragment, Fab'-SH fragment, F(ab')2 fragment, Fv fragment, single-chain Fv (scFv), dsFv or Fd fragment.

[0157] In some embodiments, the antibody or antigen-binding fragment of the present disclosure is capable of binding B7-H3 (e.g., human, monkey, or mouse B7-H3) at an EC50 of less than about 150 ng / mL, such as less than about 140 ng / mL, 130 ng / mL, 120 ng / mL, 110 ng / mL, 100 ng / mL, 90 ng / mL, 80 ng / mL, 70 ng / mL, 60 ng / mL, 50 ng / mL, 20 ng / mL, or less.

[0158] In some embodiments, the antibodies or antigen-binding fragments of this disclosure possess good endocytic activity, capable of inducing the internalization of B7-H3 (e.g., human, monkey, or mouse B7-H3), and / or, the antibodies or antigen-binding fragments or conjugates of this disclosure can enter cancer cells via endocytosis. In some embodiments, the mutated antibodies or antigen-binding fragments provided in this disclosure have significantly lower HIC values ​​compared to unmutated antibodies (e.g., 2#8890).

[0159] In some embodiments, the antibodies or antigen-binding fragments of the present disclosure have pH-dependent binding characteristics. In some embodiments, the dissociation rate of the antibodies or antigen-binding fragments of the present disclosure is higher under acidic conditions than at neutral pH. In some embodiments, the antibodies or antigen-binding fragments of the present disclosure bind B7-H3 (e.g., human, monkey, or mouse B7-H3) with a higher affinity at neutral pH than at acidic pH.

[0160] In some embodiments, the mutated antibody or its antigen-binding fragment provided in this disclosure has a higher dissociation rate under acidic conditions than the unmutated antibody, and has a longer half-life, higher recycling rate, improved pharmacokinetic properties and / or higher bioavailability compared to the unmutated antibody (such as 2#8890).

[0161] In some embodiments, the antibodies or antigen-binding fragments of this disclosure are capable of specifically binding to B7-H3 in humans or monkeys (e.g., rhesus macaques (Macaca mulatta)).

[0162] In some embodiments, the antibodies or antigen-binding fragments of this disclosure do not bind or substantially do not bind to B7-1, B7-2, B7-H1, B7-H2 and / or B7-H4.

[0163] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein do not have ADCC activity and / or do not have CDC activity.

[0164] In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein has reduced ADCC activity and / or reduced CDC activity.

[0165] According to another aspect of this disclosure, a chimeric antigen receptor is provided, the chimeric antigen receptor comprising an antigen-binding domain, a transmembrane domain and an intracellular signal transduction domain, wherein the antigen-binding domain comprises the antibody or antigen-binding fragment thereof disclosed herein.

[0166] According to another aspect of this disclosure, a multispecific antibody or antigen-binding fragment thereof is provided, the multispecific antibody or antigen-binding fragment thereof comprising two or more (e.g., three or four) antigen-binding domains, wherein one antigen-binding domain comprises the antibody or antigen-binding fragment thereof disclosed herein.

[0167] According to another aspect of this disclosure, isolated nucleic acid molecules are provided, comprising nucleotide sequences encoding the antibodies or antigen-binding fragments thereof described above, the chimeric antigen receptors thereof, the multispecific antibodies or antigen-binding fragments thereof.

[0168] Those skilled in the art will understand that nucleotides in nucleic acid molecules can be substituted based on codon degeneracy. In some embodiments, the nucleotide sequence of the nucleic acid molecule is codon-optimized.

[0169] According to another aspect of this disclosure, a carrier comprising the aforementioned nucleic acid molecules of this disclosure is provided.

[0170] In some embodiments, the vector may be an expression vector or a cloning vector. In some embodiments, the expression vector may include eukaryotic cell expression vectors and / or prokaryotic cell expression vectors. In some embodiments, the eukaryotic expression vector includes, for example, but not limited to, yeast expression vectors, mammalian expression vectors, and insect expression vectors. For example, the vector may include, but is not limited to, plasmids, kinases, retroviral vectors, lentiviral vectors, bacteriophage vectors, adenovirus vectors, adeno-associated vectors, or herpes simplex vectors.

[0171] In some embodiments, the carrier may be selected from nanoparticles, liposomes, exogenous bodies, microbubbles, or gene guns.

[0172] According to another aspect of this disclosure, a host cell is provided, comprising the antibody or antigen-binding fragment thereof of the present disclosure, the chimeric antigen receptor thereof, the multispecific antibody or antigen-binding fragment thereof, or the nucleic acid molecule thereof.

[0173] In some embodiments, the cell can be a host cell conventionally used in the art, as long as the expression vector stably expresses the carried nucleic acid molecule as the antibody or antigen-binding fragment thereof disclosed herein, chimeric antigen receptor, or multispecific antibody or antigen-binding fragment thereof. In some embodiments, the host cell can be a prokaryotic cell and / or a eukaryotic cell. The prokaryotic cell may include, for example, *Escherichia coli*, and the eukaryotic cell may include, for example, CHO cells, HEK293 cells, BHK cells, NSO cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, HeLa cells, Vero cells, Expi293 cells, hybridoma cells, yeast cells, insect cells, and plant cells.

[0174] In some embodiments, the cells may be immune cells. In some embodiments, the immune cells may include, but are not limited to, T cells, NK cells, dendritic cells (DCs), and macrophages. In these embodiments, the immune cells may express the chimeric antigen receptors described above in this disclosure.

[0175] According to another aspect of this disclosure, a method for preparing the antibody or antigen-binding fragment thereof of this disclosure is provided, the method comprising culturing the host cell of this disclosure under conditions allowing expression of the antibody or antigen-binding fragment thereof, and collecting the antibody or antigen-binding fragment thereof from the cultured host cell culture.

[0176] According to another aspect of this disclosure, a conjugate is provided, comprising the antibody or antigen-binding fragment thereof described above; and a conjugation portion.

[0177] In some implementations, the coupling portion may include, but is not limited to, a detectable marker or a therapeutic agent.

[0178] In some embodiments, the detectable marker can be any substance detectable by means of fluorescence, spectroscopy, photochemistry, biochemistry, immunology, electricity, optics, chemistry, etc. Such markers are well known in the art, and examples include, but are not limited to, enzymes (e.g., horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radionuclides (e.g., 3H, 125I, 35S, 14C, or 32P), fluorescent dyes (e.g., fluorescein isothiocyanate (FITC), fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin (PE), Texas red, rhodamine, quantum dots, or cyanine dye derivatives (e.g., Cy7, Alexa 750)), acridine esters, magnetic beads, calorimetric markers such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) microbeads, and biotin for binding avidin (e.g., streptavidin) modified with the above markers. In some embodiments, such markers are suitable for immunological assays (e.g., enzyme-linked immunosorbent assay, radioimmunoassay, fluorescence immunoassay, chemiluminescence immunoassay, etc.). In some embodiments, the detectable marker is selected from radioactive isotopes, fluorescent substances, luminescent substances, colored substances, or enzymes. In some embodiments, the detectable markers described above can be linked to the antibodies or antigen-binding fragments of this disclosure using linkers of different lengths to reduce potential steric hindrance.

[0179] In some embodiments, the detectable marker may include, but is not limited to, enzymes (e.g., horseradish peroxidase), radionuclides, fluorescent dyes, luminescent substances (e.g., chemiluminescent substances), colored substances, biotin, etc.

[0180] In some embodiments, the drug may include, for example, but not limited to, chemotherapeutic agents, immunosuppressants, and cytotoxic drugs.

[0181] In some embodiments, the coupling portion is selected from substances that can improve the biological properties of the antibody (e.g., increase serum half-life), such as chemical groups, such as polyethylene glycol (PEG), methyl, ethyl, or glycosyl groups.

[0182] According to another aspect of this disclosure, a pharmaceutical composition is provided comprising: the antibody or antigen-binding fragment thereof of the present disclosure, the chimeric antigen receptor thereof, the multispecific antibody or antigen-binding fragment thereof, the nucleic acid molecule thereof, the carrier thereof, the host cell thereof, or the conjugate thereof; and a pharmaceutically acceptable carrier thereof.

[0183] In some embodiments, the pharmaceutical composition may also include additional pharmaceutically active agents.

[0184] In some embodiments, the additional pharmaceutically active agent may be a biologically active drug, such as a drug capable of treating diseases or conditions related to B7-H3. In some embodiments, the additional pharmaceutically active agent may be selected from, but is not limited to: B7-H3 inhibitors, EGFR inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, IGFR-1 inhibitors, mTOR inhibitors, PI3 kinase inhibitors, c-met or VEGF inhibitors, chemotherapeutic agents, or any combination thereof.

[0185] In some embodiments, the antibody or its antigen-binding fragment is provided as a separate component or as a mixed component with the additional pharmaceutically active agent.

[0186] In some embodiments, the pharmaceutical composition can be administered via, for example, parenteral, subcutaneous, sublingual, rectal, nasal, intravenous, intramuscular, oral, ocular, or topical routes.

[0187] In some embodiments, the pharmaceutical composition is in the form of, for example, an aqueous solution, suspension, powder, tablet, capsule, granule, powder, pill, disintegrant, syrup, spray, gel, emulsion, injection, elixir, lozenge, suppository, etc.

[0188] According to another aspect of this disclosure, a diagnostic or therapeutic kit is provided, the kit comprising the antibody or antigen-binding fragment thereof of the present disclosure, the chimeric antigen receptor thereof, the multispecific antibody or antigen-binding fragment thereof, the nucleic acid molecule thereof, the vector thereof, the host cell thereof, the conjugate thereof, or the pharmaceutical composition thereof.

[0189] In an optional implementation, the kit may also include instructions and / or a delivery device.

[0190] In some embodiments, the kit can be used to diagnose diseases or conditions related to B7-H3.

[0191] In some embodiments, the kit can be used to prevent or treat diseases or conditions associated with B7-H3.

[0192] According to another aspect of this disclosure, the antibodies or antigen-binding fragments thereof, the chimeric antigen receptors, the multispecific antibodies or antigen-binding fragments thereof, the nucleic acid molecules, the vectors, the host cells, the conjugates, or the pharmaceutical compositions thereof are provided for inhibiting the expression of B7-H3 protein in cells or blocking the B7-H3 signaling pathway in cells.

[0193] According to another aspect of this disclosure, the use of the antibodies or antigen-binding fragments thereof, the chimeric antigen receptors, the multispecific antibodies or antigen-binding fragments thereof, the nucleic acid molecules, the carriers, the host cells, the conjugates, or the pharmaceutical compositions thereof in the preparation of medicaments for inhibiting cell (e.g., cells expressing B7-H3 protein, such as tumor cells) proliferation, or for preventing or treating diseases or conditions related to B7-H3.

[0194] In some embodiments, the antibody or its antigen-binding fragment, isolated nucleic acid molecule, carrier, host cell, conjugate, multispecific antibody, or pharmaceutical composition is administered in combination with additional pharmaceutically active agents, such as simultaneously, separately, or sequentially.

[0195] In some embodiments, the additional pharmaceutically active agent is a biologically active drug, such as a drug capable of treating diseases or conditions associated with B7-H3.

[0196] In some embodiments, the additional pharmaceutically active agent is selected from: B7-H3 inhibitors, EGFR inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, IGFR-1 inhibitors, mTOR inhibitors, PI3 kinase inhibitors, c-met or VEGF inhibitors, chemotherapeutic agents, or any combination thereof.

[0197] In some embodiments, the disease or condition associated with B7-H3 may be a tumor, such as, but not limited to, breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, thyroid cancer, or any combination thereof.

[0198] According to another aspect of this disclosure, a method for inhibiting cell proliferation is provided, the method comprising contacting the cells with the antibody or antigen-binding fragment thereof disclosed herein, the chimeric antigen receptor, the multispecific antibody or antigen-binding fragment thereof, the nucleic acid molecule, the carrier, the host cell, the conjugate, or the pharmaceutical composition thereof.

[0199] In some embodiments, the cells may be in vitro. In some embodiments, the cells may be in vivo in a subject, wherein the subject suffers from a disease or condition related to B7-H3. In some embodiments, the cells may be tumor cells.

[0200] According to another aspect of this disclosure, a method for preventing or treating diseases or conditions related to B7-H3 is provided, the method comprising administering to a subject in need an effective amount of the antibody or antigen-binding fragment thereof of the present disclosure, the chimeric antigen receptor, the multispecific antibody or antigen-binding fragment thereof, the nucleic acid molecule, the carrier, the host cell, the conjugate, or the pharmaceutical composition thereof.

[0201] In some embodiments, the method may further include administering a second therapy to the subject, the second therapy being selected from surgery, chemotherapy, radiotherapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, viral therapy, adjuvant therapy, and any combination thereof.

[0202] In an optional embodiment, the second therapy may be applied simultaneously, separately, or sequentially with the antibodies or antigen-binding fragments thereof disclosed herein, the chimeric antigen receptors, the multispecific antibodies or antigen-binding fragments thereof, the nucleic acid molecules, the carriers, the host cells, the conjugates, or the pharmaceutical compositions thereof.

[0203] In some embodiments, the disease or condition associated with B7-H3 may be a tumor, such as, but not limited to, breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, thyroid cancer, or any combination thereof.

[0204] According to another aspect of this disclosure, a method for detecting the presence or level of B7-H3 in a sample is provided, the method comprising contacting the sample with the antibody or antigen-binding fragment of this disclosure, allowing the antibody or antigen-binding fragment thereof to form a complex with B7-H3, and detecting the formation of the complex. In such an embodiment, the method can be used to diagnose diseases or conditions related to B7-H3. In such an embodiment, the method may include the steps of: detecting the presence or level of B7-H3 in a sample from a subject and comparing it with a reference value (e.g., from a healthy subject); wherein an increase in the expression level compared to the reference value indicates that the subject has a disease or condition related to B7-H3.

[0205] According to another aspect of this disclosure, the use of the antibodies or antigen-binding fragments thereof described herein in the preparation of a diagnostic kit for detecting the presence or level of B7-H3 in a sample, or for diagnosing diseases or conditions related to B7-H3.

[0206] In some embodiments, the disease or condition associated with B7-H3 may be a tumor, such as, but not limited to, breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, thyroid cancer, or any combination thereof. Attached Figure Description

[0207] Figure 1: Results of endocytic activity assay of the 2#8890 mutant antibody with human breast ductal carcinoma HCC-1954 cells. Detailed Implementation

[0208] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention in any way. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concepts of this disclosure. Such structures and techniques have also been described in many publications.

[0209] definition

[0210] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0211] Unless the context clearly indicates otherwise, the terms “a” and “an” as used herein include plural references. For example, reference to “a cell” includes multiple such cells and equivalents known to those skilled in the art, etc.

[0212] As used herein, the term "about" indicates a range of ±20% of the following value. In some embodiments, the term "about" indicates a range of ±10% of the following value. In some embodiments, the term "about" indicates a range of ±5% of the following value.

[0213] Those skilled in the art will understand that when a range of values ​​is listed herein, it is intended to indicate that any intermediate value or range of values ​​within the listed range is also within the scope of this disclosure.

[0214] As used in this article, "antibody" refers to a globulin produced by the immune system in response to antigen stimulation, resulting from the proliferation and differentiation of B lymphocytes into plasma cells. Antibodies specifically bind to corresponding antigens and mediate immune effects. They are primarily found in serum and body fluids and are important immune molecules mediating humoral immunity. The term "antibody" can encompass various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific, trispecific, or tetraspecific antibodies), single-chain molecules, and antigen-binding fragments. The chemical basis of antibodies is immunoglobulin (Ig).

[0215] As used herein, the term "monoclonal antibody" refers to antibodies derived from a substantially homogeneous group of antibodies, meaning that, apart from possible trace amounts of variant antibodies (e.g., containing naturally occurring mutations or generated during the production of the monoclonal antibody formulation, typically present in small quantities), the individual antibodies within the group are identical and / or bind to the same epitopes. Unlike polyclonal antibody formulations, which typically comprise different antibodies targeting different antigenic determinants (epitaxes), each monoclonal antibody in a monoclonal antibody formulation targets a single determinant on the antigen.

[0216] As used herein, the term "multispecific antibody" is used in its broadest sense to encompass antibodies exhibiting multi-epitope specificity. These multispecific antibodies include, but are not limited to: antibodies comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH-VL unit exhibits multi-epitope specificity; antibodies having two or more VL and VH regions, each VH-VL unit binding to a different target or a different epitope of the same target; antibodies having two or more single variable regions, each single variable region binding to a different target or a different epitope of the same target; full-length antibodies, antibody fragments, bispecific antibodies, and trispecific antibodies, antibody fragments covalently or non-covalently linked, etc.

[0217] The terms “full-length antibody” and “intact antibody” used herein are used interchangeably to refer to antibodies that are structurally similar to natural antibodies. “Natural antibody” refers to a naturally occurring immunoglobulin molecule. For example, natural IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 Daltons, composed of two light chains and two heavy chains linked by disulfide bonds. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH) (also called a variable heavy chain domain or heavy chain variable domain) and three constant domains (CH1, CH2, and CH3) (also called heavy chain constant regions). From the N-terminus to the C-terminus, each light chain has a variable region (VL) (also called a variable light chain domain or light chain variable domain) and a light chain constant domain (CL) (also called light chain constant regions). The heavy chain of an antibody can be one of five types: α (IgA), δ (IgD), ε (IgE), γ (IgG), or μ (IgM), and can be further subdivided into subtypes such as γ1 (IgG1), γ2 (IgG2), γ3 (IgG3), γ4 (IgG4), α1 (IgA1), and α2 (IgA2). The light chain of an antibody, based on the amino acid sequence of its constant domain, can be one of two types: κ light chain and λ light chain.

[0218] Within the light and heavy chains, variable and constant regions are linked by a "J" region containing approximately 12 or more amino acid residues, and the heavy chain also contains a "D" region containing approximately 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of one domain, CL. The constant region of an antibody mediates the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0219] The term "Fd fragment" as used herein refers to an antibody fragment consisting of VH and CH1 domains. The term "dAb fragment" as used herein refers to an antibody fragment consisting of a VH domain (Ward et al., Nature 341:544546 (1989)). The term "Fab fragment" as used herein refers to an antibody fragment consisting of VL, VH, CL, and CH1 domains. The term "F(ab')2 fragment" as used herein refers to an antibody fragment containing two Fab fragments linked by disulfide bridges on the hinge region. The term "Fab' fragment" as used herein refers to the fragment obtained by reducing the disulfide bonds connecting the two heavy chain fragments in the F(ab')2 fragment, consisting of a complete light and heavy chain Fd fragment (composed of VH and CH1 domains). The term "Fab'-SH" as used herein refers to a Fab fragment containing free thiol groups.

[0220] As used in this article, the term "Fv fragment" refers to an antibody fragment consisting of the VL and VH domains of a single arm of the antibody. The Fv fragment is generally considered to be the smallest antibody fragment capable of forming a complete antigen-binding site. It is generally believed that six CDRs confer antigen-binding specificity to the antibody. However, even a variable region (such as the Fd fragment, which contains only three antigen-specific CDRs) can recognize and bind to the antigen, although its affinity may be lower than that of a complete binding site.

[0221] As used herein, the term "scFv" refers to a single polypeptide chain containing VL and VH domains linked by a linker. In some cases, a disulfide bond may also exist between the VH and VL domains of the scFv.

[0222] As used herein, the terms "linker" or "connector" refer to a (peptide) connector of natural and / or synthetic origin, composed of linear amino acids. The domains in the bispecific fusion polypeptide of the present invention can be linked by linkers, wherein each linker is fused to and / or otherwise linked (e.g., via peptide bonds) with at least two polypeptides or domains. In some embodiments, all linkers present in the bispecific fusion polypeptide of the present invention have the same amino acid sequence. In other embodiments, at least two linkers present in the bispecific fusion polypeptide of the present invention have different amino acid sequences. The linker should have a length suitable for linking two or more monomeric domains in this manner, and the linker should ensure that the different domains to which it is linked fold correctly and are properly presented to perform their biologically active functions. In various embodiments, the linker has a flexible conformation. Suitable flexible linkers include, for example, those having glycine, glutamine, and / or serine residues. In some embodiments, the linker may be selected from (Glycine, glutamine, and / or serine residues). n S) m (G) nOr (EAAAK) n , where n and m are each independently selected from integers from 0 to 5. For example, n is selected from 0, 1, 2, 3, 4 or 5, and m is selected from 1, 2, 3, 4 or 5.

[0223] As used herein, the term "variable region" or "variable domain" refers to the domain of the antibody heavy or light chain involved in the binding of the antigen-binding molecule to the antigen. The variable domains (VH and VL, respectively) of the heavy and light chains of natural antibodies typically have similar structures, with each domain containing four conserved framework regions (FR1-4) and three hypervariable regions (HVR1-3), arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A single VH or VL domain is sufficient to confer antigen-binding specificity. The three HVRs within the VH and VL together constitute the antigen-binding site of Ig, which can bind complementary to the corresponding antigenic epitope; therefore, the HVRs are also called complementarity-determining regions (CDRs), denoted as CDR1, CDR2, and CDR3, respectively. The VH or VL chain of the antibody may further contain all or part of the constant regions of the heavy or light chain.

[0224] The CDR of the antibody or antigen-binding fragment thereof disclosed herein can be determined according to various numbering systems known in the art. In some embodiments, the CDR contained in the antibody or antigen-binding fragment thereof disclosed herein is preferably determined by the IMGT, Kabat, Chothia, or AbM numbering system.

[0225] The term "antibody" also includes embodiments in which the heavy chain constant region contains a C-terminal lysine, or lacks a C-terminal lysine, or a C-terminal glycine-lysine dipeptide. The term also includes embodiments in which the N-terminal amino acid of the antibody variable region has been cyclized into a pyroglutamate salt. Therefore, in compositions comprising the antibodies disclosed herein or antigen-binding fragments thereof, the various antibodies therein may independently contain a C-terminal lysine, lack a C-terminal lysine, lack a C-terminal glycine-lysine, and / or contain N-terminal glutamine or glutamate, or have the N-terminal amino acid cyclized into pyroglutamate.

[0226] As used in this paper, the term "variable" refers to the fact that certain segments of the variable region are generally different in sequence between antibodies. The V domain mediates antigen binding and defines the specificity of a particular antibody for its specific antigen. However, variability is not uniformly distributed throughout the variable region, but is concentrated in three segments called hypervariable regions (HVRs) within the variable regions of the light and heavy chains. The relatively highly conserved portions of the variable region are called framework regions (FRs). The variable regions of the native heavy and light chains each contain four FRs, mostly in a β-sheet configuration, linked by three HVRs that form loops and, in some cases, form part of a β-sheet structure. The HVRs in each chain are tightly held together by the FRs and, together with the HVRs of other chains, contribute to the formation of the antibody's antigen-binding site. Constant domains do not directly participate in antibody-antigen binding but have other effector functions, such as involvement in antibody-dependent cytotoxicity.

[0227] Antibody "classes" refer to the types of constant structural domains or constant regions possessed by the antibody's heavy chain. Based on differences in heavy chain structure and antigenicity, they can be classified into five classes: μ chain, γ chain, α chain, δ chain, and ε chain. Immunoglobulins composed of different heavy and light chains are respectively called IgA, IgD, IgE, IgG, and IgM. Even within the same class of Ig, the amino acid composition of the hinge region and the number and position of disulfide bonds in the heavy chain differ, thus allowing for further subclassing of the same class of Ig. For example, human IgG can be divided into IgG1–IgG4; IgA can be divided into IgA1 and IgA2. Based on differences in light chain structure and antigenicity, immunoglobulin (Ig) light chains are divided into κ (kappa) chains and λ (lambda) chains, thus classifying Ig into two types: κ type and λ type.

[0228] "Humanized antibodies" comprise amino acid residues from non-human HVRs and amino acid residues from human FRs. In some embodiments, humanized antibodies comprise at least one, typically two, variable domains, wherein all or substantially all HVRs (e.g., CDRs) correspond to the HVRs of the non-human antibody, and all or substantially all FRs correspond to the FRs of the human antibody. Humanized antibodies may optionally comprise at least a portion of the antibody constant region derived from a human antibody. Antibodies in a "humanized form," such as non-human antibodies, refer to antibodies that have undergone humanization.

[0229] As used herein, the term "Fc domain" or "Fc region" is used to define a C-terminal region of an antibody heavy chain containing at least a portion of a constant region. This term includes native sequence Fc regions and variant Fc regions. An IgG Fc region contains an IgG CH2 domain and an IgG CH3 domain. The CH2 domain as used herein may be a native sequence CH2 domain or a variant CH2 domain. The CH3 region as used herein may be a native sequence CH3 domain or a variant CH3 domain. The CH2 domain may contain one or more mutations that reduce or eliminate the binding of the CH2 domain to one or more Fcγ receptors (e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIII) and / or complement. It is presumed that reducing or eliminating binding to Fc receptor γ will reduce or eliminate antibody-mediated ADCC. Similarly, reducing or eliminating binding to complement is expected to reduce or eliminate antibody-mediated CDC. Mutations that reduce or eliminate the binding of the CH2 domain to one or more Fcγ receptors and / or complement are known in the art. These mutations include the so-called "LALA mutation," which involves replacing leucine residues at positions 1.3 and 1.2 of the IMGT site in the CH2 domain with alanine (L1.3A and L1.2A). Alternatively, it is also known to generate α-glycosylated antibodies by mutating asparagine (N) at position 84.4 of the IMGT site in the CH2 domain to alanine, glycine, or glutamine (N84.4A, N84.4G, or N84.4Q), thereby mutating conserved N-chain glycosylation sites to reduce IgG1 effector function. As another alternative, complement activation (C1q binding) and ADCC are known to be reduced by mutating proline at position 114 of the IMGT site in the CH2 domain to alanine or glycine (P114A or P114G). These mutations can be combined to produce antibody molecules with further reduced or no ADCC or CDC activity.

[0230] "Regions equivalent to the Fc region of immunoglobulins" include variants of the naturally occurring alleles of the immunoglobulin Fc region, as well as modified variants capable of producing substitutions, additions, or deletions that substantially do not diminish the immunoglobulin-mediated effector functions (such as antibody-dependent cytotoxicity). For example, one or more amino acids can be deleted from the N-terminus or C-terminus of the immunoglobulin Fc region without substantially losing biological function. Such variants can be selected according to general rules known in the art to minimize the impact on activity.

[0231] As used in this article, the term "effector function" refers to the biological activity attributable to the Fc region of an antibody, which varies with antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), cytokine secretion, antigen uptake by immune complex-mediated antigen-presenting cells, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0232] As used herein, the terms “polynucleotide,” “nucleic acid,” or “nucleotide sequence” refer to isolated nucleic acid molecules or constructs, such as messenger RNA (mRNA), virus-derived RNA, or plasmid DNA (pDNA). Polynucleotides may contain conventional phosphodiester bonds or unconventional bonds (such as amide bonds, as found in peptide nucleic acids (PNAs)). The term “nucleic acid molecule” refers to any one or more nucleic acid segments, such as DNA or RNA fragments, present in a polynucleotide.

[0233] An "antibody fragment" or "antigen-binding fragment" contains a portion of a complete antibody that retains the antibody's antigen-binding activity. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv; bisomatic antibodies, trisomatic antibodies, tetrasomatic antibodies, cross-Fab fragments; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments and single-domain antibodies (single-domain antibodies).

[0234] As used herein, the term "antigen-binding domain" or "antigen-binding site" refers to the portion of an antigen-binding molecule that specifically binds to an antigenic determinant. More specifically, the term "antigen-binding domain" refers to a portion of an antibody containing a region that specifically binds to and is complementary to a portion or all of the antigen. In cases where the antigen molecule is large, the antigen-binding molecule may bind only a specific portion of the antigen, called an epitope. The antigen-binding domain may be provided by, for example, one or more variable domains (also called variable regions). Preferably, the antigen-binding domain comprises a variable region (VL) of the antibody light chain and a variable region (VH) of the antibody heavy chain. In one aspect, the antigen-binding domain is capable of binding its antigen and blocking or partially blocking the function of said antigen.

[0235] As used herein, the term "antigenic determinant" is synonymous with "antigen" and "epitope" and refers to a site on a polypeptide macromolecule (e.g., a continuous amino acid sequence or a conformation composed of different regions of non-continuous amino acids) to which an antigen-binding moiety binds, thereby forming an antigen-binding moiety-antigen complex. Antigenic determinants can be present, for example, on the surface of tumor cells, on the surface of microbially infected cells, on the surface of other diseased cells, on the surface of immune cells, in serum, and / or in the extracellular matrix (ECM). Unless otherwise stated, proteins used as antigens in this invention can be any naturally occurring form of protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). Antigens can also be human proteins, or antigens can be "full-length," unprocessed proteins, and any form of protein produced by intracellular processing, or naturally occurring protein variants, such as splice variants or allelic variants.

[0236] The specific “binding strength” or “affinity” of an antibody or its antigen-binding fragment to an antigen refers to the strength of the non-covalent interaction between a single binding site and its binding ligand (e.g., antigen), and can be distinguished from unwanted or non-specific binding. The ability of an antigen-binding molecule to bind to a specific antigen can be measured by enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to those skilled in the art, such as surface plasmon resonance (SPR) techniques and conventional binding assays. In one embodiment, for example, as measured by SPR, the degree of binding of the antigen-binding molecule to an unrelated protein is less than about 10% of the degree of binding of the antigen-binding molecule to the antigen. Binding affinity is typically expressed as a dissociation constant (KD), which is the ratio of the dissociation rate constant to the association rate constant (Kd and Ka, respectively). In some embodiments, the dissociation constant (Kd) of the molecule binding to the antigen is ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10). -7 M or lower, such as 10 -7 M to 10 -13 M, for example, 10 -9 M to 10 -13 M).

[0237] As used herein, the term "isolated" nucleic acid molecule or polynucleotide refers to a nucleic acid molecule, DNA, or RNA, that has been separated from its natural environment. In this invention, the recombinant polynucleotide encoding a polypeptide contained in the vector is also isolated. Other examples of isolated polynucleotides include recombinant polynucleotides in heterologous host cells or polynucleotides purified in solution. Isolated polynucleotides include polynucleotide molecules typically found in cells containing the polynucleotide molecule, but which are located extrachromosomally or at chromosomal locations different from their natural chromosomal locations. Isolated RNA molecules include in vivo or in vitro RNA transcripts of this invention, in positive and negative strand forms, and in double strand forms. The isolated polynucleotides or nucleic acids of this disclosure further include synthetically generated molecules of this type. Additionally, the polynucleotide or nucleic acid may be or may include regulatory elements such as promoters, ribosome binding sites, or transcription terminators.

[0238] As used herein, the terms "vector" or "expression vector" and "expression construct" are used interchangeably to refer to a DNA molecule to which a specific gene, operatively linked, is introduced into a target cell and directed for expression. The vector includes vectors that function as self-replicating nucleic acid structures and vectors incorporated into the genome of the host cell into which they have been introduced. The expression vectors of the present invention comprise expression cassettes. Expression vectors can be transcribed into large quantities of stable mRNA. Once the expression vector is within the target cell, cellular transcription and / or translation mechanisms generate a ribonucleic acid molecule or protein encoded by the gene. In one embodiment, the expression vector of the present invention comprises an expression cassette containing a polynucleotide sequence encoding a fragment of the bispecific antigen-binding molecule or thereof of the present invention. The term "expression cassette" of the present invention refers to a recombinant or synthetically generated polynucleotide having a series of nucleic acid elements that allow a specific nucleic acid to be transcribed in a target cell. Recombinant expression cassettes can be introduced into plasmids, chromosomes, mitochondrial DNA, plastid DNA, viruses, or nucleic acid fragments. Typically, in addition to other sequences, the recombinant expression cassette portion of the expression vector includes the nucleic acid sequence to be transcribed and a promoter. In some embodiments, the expression cassette of the present invention comprises a polynucleotide sequence encoding a bispecific antigen-binding molecule or thereof of the present invention.

[0239] As used herein, the terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells in which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include “transformants” and “transformed cells,” including primary transformed cells and their derived progeny. The nucleic acids of the progeny may not be completely identical to those of the parent cells and may contain mutations. Host cells are any type of cell that can be used to generate the bispecific antigen-binding molecule of this invention. Host cells include cultured cells, such as cultured mammalian cells, such as CHO cells, HEK293 cells, BHK cells, NSO cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, or hybridoma cells, yeast cells, insect cells, and plant cells, as well as cells contained within transgenic animals, transgenic plants, or cultured plant or animal tissues.

[0240] As used herein, the term "antibody-drug conjugate" or "ADC" refers to a binding protein (such as an antibody or its antigen-binding fragment) chemically linked to one or more chemical drugs. In a preferred embodiment, an ADC comprises a binding protein, a drug, and a connector linking the binding protein to the drug.

[0241] As used herein, the term "chimeric antigen receptor" or "CAR" refers to a receptor having desired antigen specificity and signal transduction domains to propagate intracellular signals upon antigen binding. For example, T lymphocytes recognize specific antigens via the interaction of T cell receptors (TCRs) with short peptides presented by class I or II major histocompatibility complex (MHC) molecules. For initial activation and clonal expansion, naïve T cells depend on antigen-presenting cells (APCs) that provide additional co-stimulatory signals. In some embodiments, monocytes and macrophages can be engineered to express, for example, chimeric antigen receptors (CARs). Modified cells can be recruited to the tumor microenvironment, where they act as potent immune effectors by infiltrating the tumor and killing target cancer cells. CARs may include antigen-binding domains, transmembrane domains, and intracellular domains. The antigen-binding domain binds to the antigen on the target cell. Examples of cell surface markers that can be used as antigens binding to the antigen-binding domain of a CAR include those associated with viruses, bacteria, parasitic infections, autoimmune diseases, and cancer cells (e.g., tumor antigens).

[0242] As used herein, the term "modified immune cell" refers to an immune cell that has been genetically modified to express a CAR. In some embodiments, the immune cell is a T cell or a cell derived therefrom. In some embodiments, the immune cell is a natural killer (NK) cell or a cell derived therefrom. In some embodiments, the immune cell is a B cell or a cell derived therefrom. In some embodiments, the immune cell is a B cell or a cell derived therefrom. In some embodiments, the immune cell is a monocyte or macrophage, or a cell derived therefrom.

[0243] An "effective amount" of a drug is the amount necessary to produce physiological changes in the cells or tissues to which it is administered. An "effective amount" includes the amount sufficient to improve or prevent the symptoms or signs of a medically diagnosed disease. An effective amount also means the amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the patient's overall health, the route and dosage of administration, and the severity of any side effects. An effective amount can be the maximum dose or administration regimen that avoids significant side effects or toxicity.

[0244] The "therapeutic effective amount" of a drug (such as a pharmaceutical composition) refers to the amount necessary to effectively achieve the desired therapeutic or preventive effect in terms of dosage, dosing intervals, and time. For example, a therapeutically effective amount of a drug eliminates, mitigates / reduces, delays, minimizes, or prevents the adverse effects of a disease.

[0245] As used herein, the terms “individual” or “subject” refer to mammals. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates, such as monkeys), rabbits, and rodents (e.g., mice and rats). Specifically, an individual or subject is a human.

[0246] As used herein, the term "pharmaceutical composition" refers to a mixture containing one or more antibodies or antigen-binding fragments of the present disclosure, along with other chemical components, such as physiological / pharmaceutical-grade carriers or excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertment of its biological activity.

[0247] As used herein, the term "pharmaceuticalally acceptable excipient" refers to a component in a pharmaceutical composition that, apart from the active ingredient, is non-toxic to the subject. Pharmaceutically acceptable excipients include, but are not limited to, buffers, stabilizers, and / or preservatives.

[0248] As used herein, the term "treatment" refers to the administration of an oral or topical therapeutic agent, such as a composition comprising any antibody of this disclosure or an antigen-binding fragment thereof, or a nucleic acid molecule encoding an antibody or an antigen-binding fragment thereof, to a patient having one or more diseases or symptoms associated with B7-H3, and the therapeutic agent having a therapeutic effect on these diseases or symptoms. Typically, the therapeutic agent is administered in a treated patient or population in an amount that effectively relieves one or more diseases or symptoms, to induce regression of such symptoms, or to inhibit the development of such symptoms to any clinically measurable extent.

[0249] As used herein, the term "prevention" refers to delaying, suppressing, or preventing the onset of B7-H3-related diseases in mammals where the initiation of cancer or tumor development has not been confirmed, but a susceptibility to cancer has been identified, for example, through genetic screening or other methods. The term also includes treating mammals with precancerous lesions to halt the progression of the precancerous lesions to malignancy or to induce their regression.

[0250] As used herein, the term "detectable marker" encompasses a marker that can be detected directly or indirectly, either attached to the antibody or the bispecific binding protein, or present independently in the kit. Suitable markers include, but are not limited to, molecules detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. Suitable markers include, but are not limited to, fluorescent dyes (e.g., GFT and its variants, FITC, TRITC, fluorescein, and rhodamine), electron-dense reagents (e.g., gold), enzymes (e.g., horseradish peroxidase (HRP), alkaline phosphatase (AP), glucose oxidase, β-D-galactosidase, urease, catalase, or glucosylamylase), molecules containing radionuclides (i.e., radioisotopes), chemiluminescent molecules, electrochemiluminescent molecules, biotin, digoxin / digoxigenin, or haptens, and other entities that are or can be detectable. Antibodies or their antigen-binding fragments or bispecific binding proteins in this disclosure are attached to a "detectable marker," thus being detectably labeled.

[0251] The "sequence identity percentage" or "identity percentage" between two polynucleotide or polypeptide sequences refers to the number of identical matching positions shared by sequences within a comparison window, taking into account additions or deletions (i.e., vacancies) that must be introduced for optimal alignment of the two sequences. A matching position is any location where the same nucleotide or amino acid is present in both the target and reference sequences. Vacancies are not nucleotides or amino acids and are not counted in the target sequence. Similarly, vacancies in the reference sequence are not counted because nucleotides or amino acids from the target sequence are included, but those from the reference sequence are excluded.

[0252] The percentage of sequence identity can be calculated as follows: determine the number of positions in both sequences where the same amino acid residue or nucleic acid base appears (the number of matching positions), divide the number of matching positions by the total number of positions in the comparison window, and multiply the result by 100 to obtain the percentage of sequence identity. Sequence comparison and determination of the percentage of sequence identity between two sequences can be accomplished using software that is readily available online and downloadable. Suitable software programs are available from various sources for protein and nucleotide sequence alignment. A suitable program for determining the percentage of sequence identity is bl2seq, which is part of the BLAST program suite available from the National Center for Biotechnology Information (NCBI) website (blast.ncbi.nlm.nih.gov). Bl2seq uses either the BLASTN or BLASTP algorithm for comparing two sequences. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. Other suitable programs are, for example, Needle, Stretcher, Water, or Matcher, which are part of the EMBOSS suite of bioinformatics programs and are also available from the European Institute of Bioinformatics (EBI) at www.ebi.ac.uk / Tools / psa.

[0253] As used herein, the term "conservative substitution" refers to an amino acid substitution that does not adversely affect or alter the intended properties of a protein / peptide containing an amino acid sequence. For example, conservative substitutions can be introduced using standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include substitutions of amino acid residues with amino acid residues having similar side chains, such as substitutions with residues that are physically or functionally similar to the corresponding amino acid residues (e.g., having similar size, shape, charge, chemical properties, including the ability to form covalent or hydrogen bonds). Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid and glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, and methionine), β-branched side chains (e.g., threonine, valine, and isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine). Therefore, it is preferable to replace the corresponding amino acid residue with another amino acid residue from the same side chain family. Methods for identifying conserved amino acid substitutions are well known in the art (see, for example, Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10):879-884 (1999); and Burks et al., Proc. Natl Acad. Set USA 94:412-417 (1997), which are incorporated herein by reference).

[0254] As used herein, the term "pharmaceuticalally acceptable carrier and / or excipient" means a carrier and / or excipient that is pharmacologically and / or physiologically compatible with the subject and the active ingredient, which is well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and includes, but is not limited to: pH adjusters, surfactants, adjuvants, ionic strength enhancers, diluents, osmotic pressure maintaining agents, absorption delaying agents, and preservatives. For example, pH adjusters include, but are not limited to, phosphate buffers. Surfactants include, but are not limited to, cationic, anionic, or nonionic surfactants, such as Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Osmotic pressure maintaining agents include, but are not limited to, sugars, NaCl, and their analogues. Absorption delaying agents include, but are not limited to, monostearates and gelatin. Diluents include, but are not limited to, water, aqueous buffers (such as buffered saline), alcohols, and polyols (such as glycerol). Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, etc. Stabilizers have the meaning commonly understood by those skilled in the art for stabilizing the desired activity of the active ingredient in a pharmaceutical product, including, but not limited to, monosodium glutamate, gelatin, SPGA, sugars (such as sorbitol, mannitol, starch, sucrose, lactose, dextran, or glucose), amino acids (such as glutamic acid, glycine), proteins (such as dried whey, albumin, or casein) or their degradation products (such as lactalbumin hydrolysate), etc.

[0255] As used herein, the term "prevention" refers to a method implemented to prevent or delay the occurrence of a disease, condition, or symptom (e.g., a tumor) in a subject. As used herein, the term "treatment" refers to a method implemented to obtain a beneficial or desired clinical outcome. For the purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, alleviating symptoms, reducing the extent of the disease, stabilizing (i.e., no longer worsening) the state of the disease, delaying or slowing the progression of the disease, improving or alleviating the state of the disease, and relieving symptoms (whether partial or complete), whether detectable or undetectable. Furthermore, "treatment" can also refer to prolonged survival compared to the expected survival (if no treatment was received).

[0256] As used herein, the term "subject" refers to a mammal, such as a primate mammal, like a human. In some embodiments, the subject (e.g., a human) has a tumor, or is at risk of having the aforementioned disease.

[0257] As used herein, the term "effective amount" means an amount sufficient to achieve, or at least partially achieve, the desired effect. For example, an effective amount for preventing disease (e.g., cancer) means an amount sufficient to prevent, stop, or delay the onset of disease (e.g., cancer); an effective amount for treating disease means an amount sufficient to cure or at least partially stop the disease and its complications in a patient already suffering from the disease. Determining such an effective amount is entirely within the capabilities of those skilled in the art. For example, an effective amount for therapeutic purposes will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general characteristics such as age, weight, and sex, the manner of administration of the drug, and other concurrent treatments, etc.

[0258] As used in this article, "antibody-mediated internalization" refers to the phenomenon where an antibody, its antigen-binding fragment, or its conjugate crosses the cell membrane after binding to a cell surface antigen. Internalization includes antibody-mediated receptor internalization.

[0259] As used herein, the term "acidic pH" means a pH of 6.0 or lower (e.g., less than about 6.0, less than about 5.5, less than about 5.0, etc.). The term "acidic pH" includes pH values ​​of about 6.0, 5.95, 5.90, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0, or lower. As used herein, the term "neutral pH" means a pH of about 7.0 to about 7.4. The term "neutral pH" includes pH values ​​of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, and 7.4.

[0260] All technical features disclosed in this specification, or all steps in all disclosed methods or processes, may be combined in any way, except for mutually exclusive technical features and / or steps.

[0261] The following embodiments and accompanying drawings are provided to aid in understanding the present invention. However, it should be understood that these embodiments and drawings are for illustrative purposes only and do not constitute any limitation. The actual scope of protection of the present invention is set forth in the claims. It should be understood that any modifications and changes can be made without departing from the spirit of the present invention.

[0262] It should be noted that, unless specific conditions are specified in the examples, experimental conditions should be performed under standard conditions, manufacturer recommendations, or publicly reported conditions (e.g., refer to J. Sambrook et al., *Molecular Cloning: A Laboratory Manual (4th Edition)*, Cold Spring Harbor Laboratory). Reagents or instruments whose manufacturers are not specified are all commercially available standard products. For reagents whose manufacturers are specified, similar products from other manufacturers are substitutes.

[0263] Sequence information

[0264] The information about the sequence involved in this invention is described in Table 1 below.

[0265] Table 1. The sequences involved in this invention.

[0266] Example 1: Expression of 2#8890 mutant antibody

[0267] To improve antibody aggregation, one or more of the following mutations were used in the variable region of antibody 2#8890 (sequence derived from WO2024114525A1) to combat B7H3: heavy chain W34N, W34Y, F57H, and light chain N93D mutation. The modified antibody and 2#8890 both used the human IgG1 heavy chain constant region mutant amino acid sequence (SEQ ID NO: 45) and the human IgG1 heavy chain constant region amino acid sequence (SEQ ID NO: 35) in their heavy chain constant regions, and the light chain constant regions both used the human light chain κ constant region amino acid sequence (SEQ ID NO: 36). The amino acid sequences of the heavy and light chains of 2#8890 and its mutant antibody are shown in the table below.

[0268] Table 2.2 shows the amino acid sequences of the heavy and light chains of #8890 and its mutant antibody.

[0269] The sequences of the antibodies provided were expressed by GenScript and transfected into CHOS cells. After centrifugation, the cell supernatant was collected and purified using Protein A (MabSelect SuRe, GE).

[0270] Example 2: Cell affinity of 2#8890 mutant antibody

[0271] The affinity of the anti-human 2#8890 mutant antibody for primary ductal carcinoma HCC-1954 cells from human breast cancer patients was detected using a flow cytometer (Beckman, model Cytoflex).

[0272] HCC-1954 cells adherent to the culture vessel were digested with Tryple (Gibco, catalog number 12604013) solution. Cells were counted and a suitable amount was taken. The cells were washed twice with 1640 medium containing 1% FBS, resuspended in 1640 medium containing 1% FBS, and then transferred to 96-well conical plates, 50 μL per well. The candidate antibody was diluted with 1640 medium containing 1% FBS, starting at 20 μg / mL, and serially diluted 3-fold. 50 μL of the diluted antibody was then added to the conical plate containing the cells and incubated at 4°C for 1 h. The cells were washed twice with 1640 medium containing 1% FBS, and then 50 μL of diluted secondary antibody (Jason, Cat. 109-115-170) was added to each well. The mixture was incubated at 4°C for 30 min. The cells were washed twice with 1640 medium containing 1% FBS, and then resuspended in 100 μL of 1640 medium containing 1% FBS for flow cytometry analysis. Data processing: Median PE values ​​were exported, and EC50 was calculated. The results are shown in Table 3. The mutant antibodies 2#8890-F57H, 2#8890-F57H / N93D, 2#8890-W34N / N93D, 2#8890-W34Y / N93D, 2#8890-W34N / F57H, 2#8890-W34N / F57H / N93D, 2#8890-W34Y / F57H, and 2#8890-W34Y / F57H / N93D all showed good cell affinity.

[0273] Table 3: Affinity assay results of antibody #28890 mutant with HCC-1954 cells

[0274] Example 3: Detection of endocytic activity of the #2 8890 mutant antibody

[0275] The endocytic activity of the 2#8890 and 2#8890 mutant antibodies against primary ductal carcinoma HCC-1954 cells from human breast cancer patients was detected using flow cytometry (Beckmen, A00-1-1102-CYFOFLEX).

[0276] Adherent cells were digested with Trypsin-EDTA (0.25%) (Thermo) solution and counted. The cell density was adjusted to 1 × 10⁻⁶ cells using complete culture medium. 5 Add 100 μl of cell suspension per well to a 96-well plate (cell count 1 × 10⁶ cells / ml). 4The 96-well plate was incubated at 37°C in a CO2 incubator for 24 hours. The test antibody and negative control antibody hIgG1 were diluted with complete culture medium (Roswell Park Memorial Institute 1640 medium, Shanghai Basalmedia Technologies) to a final concentration of 8 nM, followed by 3-fold serial dilutions for a total of 7 concentration points. 300 μg / ml of pHrodo Deep Red reagent (Thermo, Cat#Z25618) was diluted with complete culture medium to 48 nM (the final concentration of pHrodo Deep Red was 24 nM). The test antibody at the high concentration points was mixed 1:1 with pHrodo Deep Red reagent and incubated at room temperature in the dark for 30 min. Then, the antibody-pHrodo Deep Red mixture was serially diluted 3-fold. The 96-well plate was removed, the culture medium was discarded, and 100 μl of the test antibody and pHrodo Deep Red reagent were taken. The Red reagent mixture was added to a 96-well plate and incubated at 37°C and 5% CO2 for 24 h. The 96-well plate was removed, the culture medium was discarded, and the plate was washed once with sterile PBS. 100 μL of Trypsin-EDTA (0.25%) was added to each well to digest the cells, followed by 100 μL of complete culture medium to neutralize them. The cells in the wells were dispersed by pipetting and then analyzed by FACS. The APC fluorescence channel gain was 230. Data processing: Median APC values ​​were exported, and EC50 was calculated. The results are shown in Figure 1 and Table 4. The 2#8890 mutant antibodies 2#8890-F57H, 2#8890-F57H / N93D, 2#8890-W34N / N93D, 2#8890-W34Y / N93D, 2#8890-W34N / F57H, 2#8890-W34N / F57H / N93D, 2#8890-W34Y / F57H, and 2#8890-W34Y / F57H / N93D exhibit good endocytic activity.

[0277] Table 4: Assay of endocytic activity of the 2#8890 mutant antibody

[0278] Example 4: Detection of 2#8890 mutant antibody by liquid-phase hydrophobic interaction chromatography (HIC)

[0279] Linear retention data of the 2#8890 mutant antibody on HIC resin (TOSOH, TSKgel Butyl-NPR) were obtained through a linear gradient experiment using a 4.6*100 mm column with pulse injection (0.1 mL protein, concentration approximately 5 mg / mL). HPLC (Agilent 1260) was used at a column temperature of 30℃, a detection wavelength of 280 nm, a flow rate of 0.5 mL / min, and mobile phases A and B: 1.5 mol / L (NH₄)₂SO₄ and 25 mmol / L Na₂HPO₄, pH 7.0, 25% IPA. An appropriate amount of the test sample was diluted with diluent (0.75 mol / L (NH₄)₂SO₄) to prepare a 1.0 mg / mL solution as the test solution. Controls 1 and 2 were diluted with diluent to prepare 1 mg / mL solutions as system suitability solutions. Inject approximately 40 μg of sample and perform analytical gradient elution: 0-3 min, maintain mobile phase A at 95% and mobile phase B at 5%; 3-40 min, increase mobile phase B from 5% to 100%; 40-45 min, maintain mobile phase A at 95% and mobile phase B at 5%. After detection, calculate the hydrophobicity value of the test sample based on the control sample. The calculation formula is as follows: (Test sample RT - Hydrophilic control RT) / (Hydrophobic control RT - Hydrophilic control RT), where RT is the peak retention time. The results are shown in Table 5. The hydrophilicity of the mutant antibodies 2#8890-F57H, 2#8890-F57H / N93D, 2#8890-W34N / N93D, 2#8890-W34Y / N93D, 2#8890-W34N / F57H, 2#8890-W34N / F57H / N93D, 2#8890-W34Y / F57H, and 2#8890-W34Y / F57H / N93D is better than that of the unmodified 2#8890 antibody.

[0280] Table 5: HIC Detection of 2#8890 Mutant Antibody

[0281] Example 5: pH-dependent detection of the 2#8890 mutant antibody

[0282] The dynamic dissociation constants of antibody 2#8890 and antibody 2#8890-F57H mutant with human B7-H3-4Ig-his protein were determined using ForteBio (Pall Life Sciences) under PBST (0.02% Tween-20) conditions at pH 7.4, pH 6, pH 5, pH 4, and pH 3.4.

[0283] The specific method is as follows: Biotinylated human B7-H3-4Ig-his protein was diluted to 100 nM with PBST (0.02% Tween-20, pH 7.4), and 2#8890 and 2#8890-F57H were diluted to 66.67 nM. Subsequently, the target biotinylated hB7-H3 protein was captured for 60 seconds in PBST (0.02% Tween-20, pH 7.4) solution using an SA Sensor (Pall Life Sciences), followed by baseline equilibration for 60 seconds. It was then bound to the parental antibody #28890 and the mutant antibody #28890-F57H for 70 seconds, respectively. Dissociation was then performed for 200 seconds in PBST (0.02% Tween-20) at pH 7.4, pH 6, pH 5, pH 4, and pH 3.4. Based on the measured results, global fitting was used to analyze the results and obtain the dynamic dissociation constant and affinity constant.

[0284] The results are shown in Table 6. The results show that the dissociation constant of the mutant antibody is larger than that of the parent antibody as the pH value decreases, which proves that the modification of histidine at the F57 site increases the dissociation rate of antigen and antibody under acidic pH conditions.

[0285] Table 6: Detection results of dynamic dissociation constant and affinity constant of human B7-H3-4Ig-his protein against 2#8890 and 2#8890-F57H mutant antibodies.

[0286] Example 6: Turbidity detection of antibodies under different stirring conditions

[0287] Take 8 mL of each of the 2#8890 antibody solution and the 2#8890-F57H-wt antibody solution (pH 5.5), both with an antibody concentration of 15 g / L, a buffer solution of 50 mmol / L acetate-sodium acetate, and a solution containing 100 mmol / L sodium chloride, and place them in 15 mL vials. Then, mix them using a magnetic stirrer (Shanghai Sile). Take another 8 mL of the same sample and place it in a 15 mL centrifuge tube, and mix it using a rotary mixer (Haimen Qilin Bell). Samples were taken at different mixing times and measured using a WGZ-1B turbidimeter (Lichen Technology). The results are shown in Tables 7 and 8 below. The results show that, under the same pH conditions, compared with the 2#8890 antibody, the modified 2#8890-F57H-wt antibody showed a significant improvement in turbidity increase under different stirring conditions. This characteristic not only enhances antibody stability and improves the operability of the production process, but also increases the efficiency of subsequent production processes (such as the preparation of antibody-drug conjugates and bispecific antibodies) and the stability of subsequent products.

[0288] Table 7: Turbidity Detection Results of Antibody #2 8890 Note: " / " indicates that it was not detected.

[0289] Table 8: Turbidity Detection Results of Antibody #28890-F57H-wt

[0290] The technical solutions disclosed herein are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of this invention fall within the protection scope of this invention.

Claims

1. An antibody or its antigen-binding fragment that specifically binds to B7-H3, wherein, The antibody or its antigen-binding fragment includes: An amino acid residue with an N or Y at position 34 in the variable region of the heavy chain; Amino acid residues with an H at position 57 in the variable region of the heavy chain; and / or The amino acid residue with a D at position 93 in the variable region of the light chain. Preferably, the positions of the amino acid residues in the heavy chain variable region are defined relative to the amino acid sequence shown in SEQ ID NO:

1. Preferably, the positions of the amino acid residues in the light chain variable region are defined relative to the amino acid sequence shown in SEQ ID NO:

7.

2. The antibody or its antigen-binding fragment according to claim 1, wherein, The antibody or its antigen-binding fragment includes: The amino acid residue with N at position 34 and the amino acid residue with F at position 57 in the variable region of the heavy chain; The amino acid residue with N at position 34 and the amino acid residue with H at position 57 in the variable region of the heavy chain; The amino acid residue with Y at position 34 and the amino acid residue with F at position 57 in the variable region of the heavy chain; The amino acid residue with Y at position 34 and H at position 57 in the variable region of the heavy chain; or The amino acid residues with W at position 34 and H at position 57 in the variable region of the heavy chain.

3. The antibody or its antigen-binding fragment according to claim 2, wherein, The antibody or its antigen-binding fragment further includes an amino acid residue with a D at position 93 in the variable region of the light chain.

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein, The antibody or its antigen-binding fragment includes: (1) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (2) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (3) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:3, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (4) Heavy chain variable region (VH), comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence shown in SEQ ID NO:3; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (5) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (6) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (7) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (8) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (9) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (10) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it, or (11) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:1, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it.

5. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, wherein, The antibody or its antigen-binding fragment further includes a constant region derived from human or mouse immunoglobulins; and / or The antigen-binding fragments of the antibody include Fab fragments, Fab' fragments, Fab'-SH fragments, F(ab')2 fragments, Fv fragments, single-chain Fv (scFv), dsFv, or Fd fragments; and / or The heavy chain and / or light chain of the antibody or its antigen-binding fragment includes C-terminal lysine, lacks C-terminal lysine, or lacks C-terminal glycine-lysine; and / or The N-terminus of the heavy chain and / or light chain of the antibody or its antigen-binding fragment includes glutamine or glutamic acid, or the N-terminal amino acid is cyclized to pyroglutamic acid or pyroglutamate.

6. The antibody according to any one of claims 1 to 5, wherein the antibody comprises: (1) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (2) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (3) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:39, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (4) A heavy chain comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence shown in SEQ ID NO:39; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (5) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (6) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (7) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (8) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (9) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (10) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (11) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:37, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (12) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (13) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (14) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:47, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (15) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:48, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (16) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (17) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (18) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it, or (19) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it.

7. An antibody or its antigen-binding fragment that specifically binds to B7-H3, wherein, The antibody or its antigen-binding fragment includes: (1) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:2; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

7. (2) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:2; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

8. (3) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:3; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

7. (4) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:3; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

8. (5) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:4; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

7. (6) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:4; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

8. (7) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:5; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

7. (8) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:5; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

8. (9) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:6; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

7. (10) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:6; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:

8. (11) CDR-H1, CDR-H2, and CDR-H3 included in the heavy chain variable region (VH) having the amino acid sequence shown in SEQ ID NO:1; and / or, CDR-L1, CDR-L2, and CDR-L3 included in the light chain variable region (VL) having the amino acid sequence shown in SEQ ID NO:8; or (12) CDR-H1, CDR-H2, and CDR-H3 having one or more amino acid substitutions, deletions, or additions compared to CDR-H1, CDR-H2, and CDR-H3 as shown in any of (1) to (11); and / or CDR-L1, CDR-L2, and CDR-L3 having one or more amino acid substitutions, deletions, or additions compared to CDR-L1, CDR-L2, and CDR-L3 as shown in any of (1) to (11), Preferably, the CDR is defined according to a numbering system of IMGT, Kabat, Chothia, AbM, or a combination thereof.

8. The antibody or antigen-binding fragment thereof according to claim 7, wherein, (i) The CDR is defined according to the IMGT numbering system. Preferably, the antibody or its antigen-binding fragment comprises: (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:15, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:15, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (5) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (6) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (7) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (8) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:16, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (9) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (10) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:17, CDR-H2 having the amino acid sequence shown in SEQ ID NO:19, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (11) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:15, CDR-H2 having the amino acid sequence shown in SEQ ID NO:18, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:20; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:9, CDR-L2 having the amino acid sequence shown in SEQ ID NO:10, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or (12) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions, or additions compared to CDR-H1, CDR-H2, and CDR-H3 as shown in (1) to (11); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions, or additions compared to CDR-L1, CDR-L2, and CDR-L3 as shown in (1) to (11), or (ii) The CDR is defined according to the Chothia numbering system. Preferably, the antibody or its antigen-binding fragment comprises: (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:23, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:23, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:22, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:21, CDR-H2 having the amino acid sequence shown in SEQ ID NO:22, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or (5) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions, or additions compared to CDR-H1, CDR-H2, and CDR-H3 as shown in (1) to (4); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions, or additions compared to CDR-L1, CDR-L2, and CDR-L3 as shown in (1) to (4), or (iii) The CDR is defined according to the Kabat numbering system. Preferably, the antibody or its antigen-binding fragment comprises: (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:25, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:25, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (5) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (6) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (7) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (8) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:26, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (9) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (10) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:27, CDR-H2 having the amino acid sequence shown in SEQ ID NO:29, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (11) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:25, CDR-H2 having the amino acid sequence shown in SEQ ID NO:28, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or (12) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions, or additions compared to CDR-H1, CDR-H2, and CDR-H3 as shown in (1) to (11); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions, or additions compared to CDR-L1, CDR-L2, and CDR-L3 as shown in (1) to (11), or (iv) The CDR is defined according to the AbM numbering system. Preferably, the antibody or its antigen-binding fragment comprises: (1) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:30, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (2) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:30, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (3) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (4) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (5) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (6) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (7) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (8) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:31, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (9) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

11. (10) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:32, CDR-H2 having the amino acid sequence shown in SEQ ID NO:34, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:

12. (11) A VH comprising the following three CDRs: CDR-H1 having the amino acid sequence shown in SEQ ID NO:30, CDR-H2 having the amino acid sequence shown in SEQ ID NO:33, and CDR-H3 having the amino acid sequence shown in SEQ ID NO:24; and / or, a VL comprising the following three CDRs: CDR-L1 having the amino acid sequence shown in SEQ ID NO:13, CDR-L2 having the amino acid sequence shown in SEQ ID NO:14, and CDR-L3 having the amino acid sequence shown in SEQ ID NO:12, or (12) A VH comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-H1, CDR-H2 and CDR-H3 as shown in (1) to (11); and / or a VL comprising the following three CDRs: having one or more amino acid substitutions, deletions or additions compared to CDR-L1, CDR-L2 and CDR-L3 as shown in (1) to (11).

9. The antibody or antigen-binding fragment thereof according to claim 7 or 8, wherein, The antibody or its antigen-binding fragment includes: (1) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (2) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:2, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (3) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:3, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (4) Heavy chain variable region (VH), comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence shown in SEQ ID NO:3; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (5) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (6) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:4, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (7) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (8) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:5, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it. (9) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:7, or an amino acid sequence having at least 80% sequence identity with it. (10) A heavy chain variable region (VH) comprising the amino acid sequence shown in SEQ ID NO:6, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it, or (11) Heavy chain variable region (VH), comprising the amino acid sequence shown in SEQ ID NO:1, or an amino acid sequence having at least 80% sequence identity with it; and / or, light chain variable region (VL), comprising the amino acid sequence shown in SEQ ID NO:8, or an amino acid sequence having at least 80% sequence identity with it.

10. The antibody or antigen-binding fragment thereof according to any one of claims 7 to 9, wherein, The antibody or its antigen-binding fragment further includes at least a portion of a constant region derived from immunoglobulins of mice, primates, cattle, horses, pigs, sheep, or goats. Preferably, the antibody or its antigen-binding fragment includes at least a portion of a constant region derived from immunoglobulins of mice, rats, lemurs, macaques, chimpanzees, or humans, more preferably from a constant region derived from human immunoglobulins. Preferably, at least a portion of the heavy chain constant region of the antibody or its antigen-binding fragment includes a heavy chain constant region derived from IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4, or IgM immunoglobulin. Preferably, the antibody or its antigen-binding fragment comprises a heavy chain constant region derived from human IgG1, or a variant thereof containing mutations or chemical modifications. Preferably, the heavy chain constant region of the antibody or its antigen-binding fragment includes the amino acid sequence shown in SEQ ID NO:35 or 45, or an amino acid sequence having at least 80% sequence identity with it, or having up to 20 conserved amino acid substitutions compared to it. Preferably, at least a portion of the light chain constant region of the antibody or its antigen-binding fragment comprises a light chain constant region derived from human κ and λ immunoglobulins, or a variant thereof containing mutations or chemical modifications. Preferably, the light chain constant region of the antibody or its antigen-binding fragment includes the amino acid sequence shown in SEQ ID NO:36, or an amino acid sequence having at least 80% sequence identity with it, or having at most 20 conserved substitutions with it. Preferably, the antibody or its antigen-binding fragment is a murine antibody, a chimeric antibody, a humanized antibody, or a fully human antibody.

11. The antibody or antigen-binding fragment thereof according to any one of claims 7 to 10, wherein, The antibody or its antigen-binding fragment includes: (1) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (2) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:38, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (3) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:39, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (4) A heavy chain comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence shown in SEQ ID NO:39; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (5) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (6) a heavy chain comprising the amino acid sequence shown in SEQ ID NO:40, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (7) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (8) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:41, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (9) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (10) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:42, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (11) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:37, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (12) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (13) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:46, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (14) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:47, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (15) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:48, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (16) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it. (17) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:49, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it. (18) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:43, or an amino acid sequence having at least 80% sequence identity with it, or (19) A heavy chain comprising the amino acid sequence shown in SEQ ID NO:50, or an amino acid sequence having at least 80% sequence identity with it; and / or a light chain comprising the amino acid sequence shown in SEQ ID NO:44, or an amino acid sequence having at least 80% sequence identity with it.

12. The antibody or antigen-binding fragment thereof according to any one of claims 7 to 11, wherein, The antigen-binding fragments of the antibody include Fab fragments, Fab' fragments, Fab'-SH fragments, F(ab')2 fragments, Fv fragments, single-chain Fv (scFv), dsFv, or Fd fragments; and / or The heavy chain and / or light chain of the antibody or its antigen-binding fragment includes C-terminal lysine, lacks C-terminal lysine, or lacks C-terminal glycine-lysine; and / or The N-terminus of the heavy chain and / or light chain of the antibody or its antigen-binding fragment includes glutamine or glutamic acid, or the N-terminal amino acid is cyclized to pyroglutamic acid or pyroglutamate.

13. Chimeric antigen receptors, among which, The chimeric antigen receptor includes an antigen-binding domain, a transmembrane domain, and an intracellular signal transduction domain, wherein the antigen-binding domain includes the antibody or its antigen-binding fragment as described in any one of claims 1 to 12.

14. A multispecific antibody or its antigen-binding fragment, wherein, The multispecific antibody or its antigen-binding fragment comprises two or more antigen-binding domains, wherein one antigen-binding domain comprises the antibody or its antigen-binding fragment as described in any one of claims 1 to 12.

15. Isolated nucleic acid molecules, among which, The nucleic acid molecule includes a nucleotide sequence encoding an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 12, a chimeric antigen receptor as described in claim 13, or a multispecific antibody or antigen-binding fragment thereof as described in claim 14.

16. Carrier, among which, The carrier comprises the nucleic acid molecule of claim 15.

17. Host cell, among which, The host cell comprises the antibody or antigen-binding fragment thereof of any one of claims 1 to 12, the chimeric antigen receptor of claim 13, the multispecific antibody or antigen-binding fragment thereof of claim 14, or the nucleic acid molecule of claim 15.

18. A method for preparing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 12, wherein, The method includes: culturing host cells and collecting the antibody or its antigen-binding fragment from the cultured host cell culture.

19. Coupled objects, among which, The conjugate comprises the antibody or antigen-binding fragment thereof as described in any one of claims 1 to 12; and, a conjugation portion.

20. The coupling according to claim 19, wherein, The coupling portion is a detectable marker or therapeutic agent. Preferably, the detectable markers include enzymes, radionuclides, fluorescent dyes, luminescent substances, colored substances, and biotin. Preferably, the therapeutic agent includes a chemotherapeutic agent, an immunosuppressant, and a cytotoxic drug.

21. A pharmaceutical composition, wherein, The pharmaceutical composition comprises: an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 12, a chimeric antigen receptor as described in claim 13, a multispecific antibody or antigen-binding fragment thereof as described in claim 14, a nucleic acid molecule as described in claim 15, a vector as described in claim 16, a host cell as described in claim 17, or a conjugate as described in claim 19 or 20; and a pharmaceutically acceptable vector.

22. The pharmaceutical composition according to claim 21, wherein, The pharmaceutical composition also includes additional pharmaceutically active agents. Preferably, the additional pharmaceutically active agent is a biologically active drug, more preferably a drug capable of treating diseases or conditions related to B7-H3. Preferably, the additional pharmaceutically active agent is selected from B7-H3 inhibitors, EGFR inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, IGFR-1 inhibitors, mTOR inhibitors, PI3 kinase inhibitors, c-met or VEGF inhibitors, chemotherapeutic agents, or any combination thereof.

23. A diagnostic or therapeutic kit, wherein the kit comprises: The antibody or antigen-binding fragment thereof of any one of claims 1 to 12, the chimeric antigen receptor of claim 13, the multispecific antibody or antigen-binding fragment thereof of claim 14, the nucleic acid molecule of claim 15, the vector of claim 16, the host cell of claim 17, the conjugate of claim 19 or 20, or the pharmaceutical composition of claim 21 or 22; and optionally, the instruction manual and / or the delivery device.

24. The antibody or antigen-binding fragment thereof of any one of claims 1 to 12, the chimeric antigen receptor of claim 13, the multispecific antibody or antigen-binding fragment thereof of claim 14, the nucleic acid molecule of claim 15, the vector of claim 16, the host cell of claim 17, the conjugate of claim 19 or 20, or the pharmaceutical composition of claim 21 or 22, are used for one or more of the following purposes: (1) Diagnose diseases or conditions related to B7-H3; (2) Prevention or treatment of diseases or conditions related to B7-H3; (3) Inhibit the expression of B7-H3 protein in cells or block the B7-H3 signaling pathway in cells; (4) Inhibit the proliferation of cells expressing B7-H3 protein, wherein the cells expressing B7-H3 protein are preferably tumor cells; (5) Detect the presence or level of B7-H3 in the sample. Preferably, the disease or condition associated with B7-H3 is a tumor, more preferably including breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, thyroid cancer, or any combination thereof.

25. Methods for preventing or treating diseases or conditions related to B7-H3, wherein, The method comprises administering to a subject in need an effective amount of any one of claims 1 to 12, an antibody or antigen-binding fragment thereof of claim 13, a chimeric antigen receptor of claim 14, a multispecific antibody or antigen-binding fragment thereof of claim 14, a nucleic acid molecule of claim 15, a vector of claim 16, a host cell of claim 17, a conjugate of claim 19 or 20, or a pharmaceutical composition of claim 21 or 22.

26. The method of claim 25, wherein, The method further includes administering a second therapy to the subject, the second therapy being selected from surgery, chemotherapy, radiotherapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, viral therapy, adjuvant therapy, and any combination thereof. Preferably, the disease or condition associated with B7-H3 is a tumor, more preferably including breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, thyroid cancer, or any combination thereof.

27. A method for detecting the presence or level of B7-H3 in a sample, wherein, The method includes: contacting the sample with an antibody or antigen-binding fragment thereof according to any one of claims 1 to 12 to allow the antibody or antigen-binding fragment thereof to form a complex with B7-H3, and detecting the formation of the complex.

28. The method according to claim 27, wherein, The method is used to diagnose diseases or conditions related to B7-H3. Preferably, the method includes the following steps: detecting the presence or level of B7-H3 in a sample from the subject and comparing it with a reference value; wherein an increase in the expression level compared to the reference value indicates that the subject has a disease or condition related to B7-H3. Preferably, the disease or condition associated with B7-H3 is a tumor, more preferably including breast cancer, colorectal cancer, head and neck cancer, clear cell renal cell carcinoma, papillary renal cell carcinoma, liver cancer, lung adenocarcinoma, lung squamous cell carcinoma, prostate cancer, gastric adenocarcinoma, thyroid cancer, or any combination thereof.

Images