Antibodies and their uses

Abstract

The present invention discloses an anti-CDCP1 antibody or an antigen-binding fragment thereof, nucleic acid molecules encoding them, and methods for producing them. The anti-CDCP1 antibody or antigen-binding fragment thereof has target binding ability, tumor cell binding ability, and endocytosis ability, and has the ability to inhibit tumor cell migration and drug development potential. The present invention also discloses a conjugate of the antibody or antigen-binding fragment thereof. The present invention also discloses a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof and its use for producing a drug, which is used to prevent and / or treat CDCP1-related diseases, such as tumors.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority from Chinese Patent Application No. 2022107001160, filed June 20, 2022. This application incorporates the above Chinese patent application in its entirety.

[0002] [Technical Field] The present invention belongs to the field of biomedicine. Specifically, the present invention relates to an antibody against CDCP1, a complex containing the same, and uses thereof, particularly for treating and / or preventing CDCP1-related diseases. [Background technology]

[0003] CUB domain-containing protein 1 (CDCP1), also known as cluster of differentiation (CD318), is a type I transmembrane glycoprotein containing an extracellular domain (ECD) with two or three CUB (Complement protein components C1, Urchin embryonic growth factor, and Bone morphogenic protein 1) domains, and an intracellular domain (ICD) containing five tyrosine residues (Y707, Y734, Y743, Y762, and Y806). There are two variants in the CDCP1 gene, the predominant one being 6.4 kb in length, located on chromosome 3p21.31, consisting of nine exons, encoding 836 amino acids, with a molecular weight of 90 kDa. The actual size measured by SDS-PAGE is 135 kDa or 140 kDa (CDCP1-135 or p140), which is thought to be related to glycosylation level. The CDCP1 extracellular domain is hydrolyzed by serine proteases near its Arg368 position to produce a 70-80 kDa (p80 or CDCP1-70) and a 65 kDa free CDCP1-65. The expression levels of p140 and p80 vary among cells, primarily determined by serine protease levels. In vivo, CDCP1 exists primarily as unhydrolyzed p140, and hydrolysis of CDCP1 occurs only upon tumor formation or tissue damage.

[0004] CDCP1 has very low homology to other proteins in the human body, with less than 20% homology to the CUB domain. Unlike CDCP1, other CUB-containing proteins lack an intracellular tyrosine domain. Furthermore, CDCP1 has high homology with mammals; for example, protein sequence analysis revealed high homology between human and mouse proteins (81.3%) and with the cynomolgus monkey protein sequence (95.8%).

[0005] CDCP1 is widely expressed in human epithelial tissue cells and hematopoietic stem and progenitor cells, but not in fibroblasts, mesenchymal stem cells, or differentiated blood cells. Multiple studies have shown that overexpression of this protein in tumors is associated with poor prognosis in kidney, lung, colorectal, pancreatic, and ovarian cancers. Phosphorylation of this protein occurs primarily in mitotic or detached cells, where it suppresses adhesion signals, disrupts focal adhesions, and negatively regulates cell adhesion. The phosphorylation level of CDCP1 is related to the cell adhesion state; its phosphorylation level increases during cell detachment, whereas its dephosphorylation level increases during cell adhesion. During cell culture, both cell division and digestion with trypsin or EDTA increase the phosphorylation level of CDCP1, and overexpression of CDCP1 leads to cell detachment or detachment. The role of CDCP1 in cell migration has been demonstrated, as both downregulation and upregulation of CDCP1 can inhibit cell migration. The main role of CDCP1 is to regulate integrin receptors; when CDCP1 is phosphorylated, it forms a complex with β1 integrin and prevents integrin aggregation, thereby disrupting the connection between the cytoskeleton and the extracellular matrix.

[0006] Analysis of clinical samples revealed elevated CDCP1 expression levels in pancreatic, colon, lung, kidney, ovarian, and breast cancers. Ki67 levels were higher in tumor cells from lung cancer patients with high CDCP1 expression than in tumor cells from patients with low CDCP1 expression. High-expression patients exhibited better tumor cell proliferation. CDCP1-p-Y734 levels were significantly elevated in lung and gastric cancer patients. Results showed that 60 and 200 lung cancer patients had moderate or high CDCP1 expression, with high-expression patients exhibiting a higher risk of lymph node metastasis and recurrence and a lower 5-year survival rate. Furthermore, a study of 230 kidney cancer patients revealed that CDCP1 expression was rarely detected in normal tissues, but high CDCP1 expression was detected in 33.5% of patient tissues. CDCP1 is also highly expressed in lymphoma cells.

[0007] CDCP1 is highly expressed in tumor tissues, making it a promising target for tumor therapy. However, there are currently no commercially available anti-CDCP1 antibody drugs. Therefore, the development of drugs against antibodies targeting CDCP1 is urgent and necessary, which will provide more options for drug use in cancer patients. Summary of the Invention

[0008] (Antibody of the present invention) In one aspect of the present invention, there is provided an antibody or antigen-binding fragment thereof that specifically binds to CDCP1, The antibody or antigen-binding fragment thereof (1) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 11, CDR-H2 comprising the sequence shown in SEQ ID NO: 48, and CDR-H3 comprising the sequence shown in SEQ ID NO: 90; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 136, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 14, CDR-H2 comprising the sequence shown in SEQ ID NO: 52, and CDR-H3 comprising the sequence shown in SEQ ID NO: 95; and a light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 140; CDR-L2 comprising the sequence shown in SEQ ID NO: 175; and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; Or, (3) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 2, CDR-H2 comprising the sequence shown in SEQ ID NO: 39, and CDR-H3 comprising the sequence shown in SEQ ID NO: 81; and a light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 125; CDR-L2 comprising the sequence shown in SEQ ID NO: 166; and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; Or, (4) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 1, CDR-H2 comprising the sequence shown in SEQ ID NO: 38, and CDR-H3 comprising the sequence shown in SEQ ID NO: 80; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 124, CDR-L2 comprising the sequence shown in SEQ ID NO: 164, and CDR-L3 comprising the sequence shown in SEQ ID NO: 197; Or, (5) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 1, CDR-H2 comprising the sequence shown in SEQ ID NO: 38, and CDR-H3 comprising the sequence shown in SEQ ID NO: 80; and a light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 125; CDR-L2 comprising the sequence shown in SEQ ID NO: 165; and CDR-L3 comprising the sequence shown in SEQ ID NO: 198; Or, (6) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 3; CDR-H2 comprising the sequence shown in SEQ ID NO: 39; and CDR-H3 comprising the sequence shown in SEQ ID NO: 81; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 126, CDR-L2 comprising the sequence shown in SEQ ID NO: 167, and CDR-L3 comprising the sequence shown in SEQ ID NO: 200; Or, (7) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 4; CDR-H2 comprising the sequence shown in SEQ ID NO: 40; and CDR-H3 comprising the sequence shown in SEQ ID NO: 82; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 127, CDR-L2 comprising the sequence shown in SEQ ID NO: 168, and CDR-L3 comprising the sequence shown in SEQ ID NO: 201; Or, (8) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 5; CDR-H2 comprising the sequence shown in SEQ ID NO: 41; and CDR-H3 comprising the sequence shown in SEQ ID NO: 83; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 128, CDR-L2 comprising the sequence shown in SEQ ID NO: 169, and CDR-L3 comprising the sequence shown in SEQ ID NO: 202; Or, (9) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 6, CDR-H2 comprising the sequence shown in SEQ ID NO: 42, and CDR-H3 comprising the sequence shown in SEQ ID NO: 84; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 129, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (10) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 6, CDR-H2 comprising the sequence shown in SEQ ID NO: 43, and CDR-H3 comprising the sequence shown in SEQ ID NO: 85; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 130, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (11) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 7, CDR-H2 comprising the sequence shown in SEQ ID NO: 44, and CDR-H3 comprising the sequence shown in SEQ ID NO: 86; and a light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 131, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 204; Or, (12) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 7, CDR-H2 comprising the sequence shown in SEQ ID NO: 44, and CDR-H3 comprising the sequence shown in SEQ ID NO: 86; and a light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 132, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 204; Or, (13) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 7, CDR-H2 comprising the sequence shown in SEQ ID NO: 44, and CDR-H3 comprising the sequence shown in SEQ ID NO: 86; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 130, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (14) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 8; CDR-H2 comprising the sequence shown in SEQ ID NO: 45; and CDR-H3 comprising the sequence shown in SEQ ID NO: 87; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 133, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (15) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 9, CDR-H2 comprising the sequence shown in SEQ ID NO: 46, and CDR-H3 comprising the sequence shown in SEQ ID NO: 88; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 134, CDR-L2 comprising the sequence shown in SEQ ID NO: 164, and CDR-L3 comprising the sequence shown in SEQ ID NO: 205; Or, (16) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 10, CDR-H2 comprising the sequence shown in SEQ ID NO: 47, and CDR-H3 comprising the sequence shown in SEQ ID NO: 89; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 135, CDR-L2 comprising the sequence shown in SEQ ID NO: 172, and CDR-L3 comprising the sequence shown in SEQ ID NO: 206; Or, (17) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 12, CDR-H2 comprising the sequence shown in SEQ ID NO: 48, and CDR-H3 comprising the sequence shown in SEQ ID NO: 91; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 136, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (18) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 11, CDR-H2 comprising the sequence shown in SEQ ID NO: 48, and CDR-H3 comprising the sequence shown in SEQ ID NO: 90; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 136, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (19) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13; CDR-H2 comprising the sequence shown in SEQ ID NO: 49; and CDR-H3 comprising the sequence shown in SEQ ID NO: 92; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 137, CDR-L2 comprising the sequence shown in SEQ ID NO: 173, and CDR-L3 comprising the sequence shown in SEQ ID NO: 208; Or, (20) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13; CDR-H2 comprising the sequence shown in SEQ ID NO: 50; and CDR-H3 comprising the sequence shown in SEQ ID NO: 93; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 138, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 209; Or, (21) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13, CDR-H2 comprising the sequence shown in SEQ ID NO: 51, and CDR-H3 comprising the sequence shown in SEQ ID NO: 94; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 139, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 210; Or, (22) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 15, CDR-H2 comprising the sequence shown in SEQ ID NO: 53, and CDR-H3 comprising the sequence shown in SEQ ID NO: 96; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 124, CDR-L2 comprising the sequence shown in SEQ ID NO: 164, and CDR-L3 comprising the sequence shown in SEQ ID NO: 212; Or, (23) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 16, CDR-H2 comprising the sequence shown in SEQ ID NO: 54, and CDR-H3 comprising the sequence shown in SEQ ID NO: 97; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 141, CDR-L2 comprising the sequence shown in SEQ ID NO: 172, and CDR-L3 comprising the sequence shown in SEQ ID NO: 213; Or, (24) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 17, CDR-H2 comprising the sequence shown in SEQ ID NO: 55, and CDR-H3 comprising the sequence shown in SEQ ID NO: 98; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 126, CDR-L2 comprising the sequence shown in SEQ ID NO: 174, and CDR-L3 comprising the sequence shown in SEQ ID NO: 214; Or, (25) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 18, CDR-H2 comprising the sequence shown in SEQ ID NO: 56, and CDR-H3 comprising the sequence shown in SEQ ID NO: 99; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 142, CDR-L2 comprising the sequence shown in SEQ ID NO: 176, and CDR-L3 comprising the sequence shown in SEQ ID NO: 215; Or, (26) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 19, CDR-H2 comprising the sequence shown in SEQ ID NO: 57, and CDR-H3 comprising the sequence shown in SEQ ID NO: 100; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 143, CDR-L2 comprising the sequence shown in SEQ ID NO: 177, and CDR-L3 comprising the sequence shown in SEQ ID NO: 216; Or, (27) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 20, CDR-H2 comprising the sequence shown in SEQ ID NO: 58, and CDR-H3 comprising the sequence shown in SEQ ID NO: 101; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 144, CDR-L2 comprising the sequence shown in SEQ ID NO: 176, and CDR-L3 comprising the sequence shown in SEQ ID NO: 215; Or, (28) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 21, CDR-H2 comprising the sequence shown in SEQ ID NO: 59, and CDR-H3 comprising the sequence shown in SEQ ID NO: 102; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 127, CDR-L2 comprising the sequence shown in SEQ ID NO: 168, and CDR-L3 comprising the sequence shown in SEQ ID NO: 217; Or, (29) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 21, CDR-H2 comprising the sequence shown in SEQ ID NO: 60, and CDR-H3 comprising the sequence shown in SEQ ID NO: 102; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 127, CDR-L2 comprising the sequence shown in SEQ ID NO: 168, and CDR-L3 comprising the sequence shown in SEQ ID NO: 217; Or, (30) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13; CDR-H2 comprising the sequence shown in SEQ ID NO: 49; and CDR-H3 comprising the sequence shown in SEQ ID NO: 103; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 144, CDR-L2 comprising the sequence shown in SEQ ID NO: 169, and CDR-L3 comprising the sequence shown in SEQ ID NO: 218; Or, (31) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 22, CDR-H2 comprising the sequence shown in SEQ ID NO: 61, and CDR-H3 comprising the sequence shown in SEQ ID NO: 104; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 145, CDR-L2 comprising the sequence shown in SEQ ID NO: 178, and CDR-L3 comprising the sequence shown in SEQ ID NO: 219; Or, (32) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 17, CDR-H2 comprising the sequence shown in SEQ ID NO: 62, and CDR-H3 comprising the sequence shown in SEQ ID NO: 105; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 141, CDR-L2 comprising the sequence shown in SEQ ID NO: 172, and CDR-L3 comprising the sequence shown in SEQ ID NO: 220; Or, (33) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 17, CDR-H2 comprising the sequence shown in SEQ ID NO: 63, and CDR-H3 comprising the sequence shown in SEQ ID NO: 106; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 126, CDR-L2 comprising the sequence shown in SEQ ID NO: 179, and CDR-L3 comprising the sequence shown in SEQ ID NO: 221; Or, (34) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 23, CDR-H2 comprising the sequence shown in SEQ ID NO: 64, and CDR-H3 comprising the sequence shown in SEQ ID NO: 107; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 146, CDR-L2 comprising the sequence shown in SEQ ID NO: 180, and CDR-L3 comprising the sequence shown in SEQ ID NO: 222; Or, (35) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 24, CDR-H2 comprising the sequence shown in SEQ ID NO: 65, and CDR-H3 comprising the sequence shown in SEQ ID NO: 108; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 147, CDR-L2 comprising the sequence shown in SEQ ID NO: 181, and CDR-L3 comprising the sequence shown in SEQ ID NO: 223; Or, (36) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 25, CDR-H2 comprising the sequence shown in SEQ ID NO: 65, and CDR-H3 comprising the sequence shown in SEQ ID NO: 108; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 148, CDR-L2 comprising the sequence shown in SEQ ID NO: 181, and CDR-L3 comprising the sequence shown in SEQ ID NO: 224; Or, (37) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 26, CDR-H2 comprising the sequence shown in SEQ ID NO: 66, and CDR-H3 comprising the sequence shown in SEQ ID NO: 109; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 149, CDR-L2 comprising the sequence shown in SEQ ID NO: 182, and CDR-L3 comprising the sequence shown in SEQ ID NO: 225; Or, (38) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 27, CDR-H2 comprising the sequence shown in SEQ ID NO: 67, and CDR-H3 comprising the sequence shown in SEQ ID NO: 110; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 150, CDR-L2 comprising the sequence shown in SEQ ID NO: 183, and CDR-L3 comprising the sequence shown in SEQ ID NO: 226; Or, (39) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 28, CDR-H2 comprising the sequence shown in SEQ ID NO: 68, and CDR-H3 comprising the sequence shown in SEQ ID NO: 111; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 151, CDR-L2 comprising the sequence shown in SEQ ID NO: 184, and CDR-L3 comprising the sequence shown in SEQ ID NO: 227; Or, (40) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 29, CDR-H2 comprising the sequence shown in SEQ ID NO: 69, and CDR-H3 comprising the sequence shown in SEQ ID NO: 112; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 152, CDR-L2 comprising the sequence shown in SEQ ID NO: 185, and CDR-L3 comprising the sequence shown in SEQ ID NO: 222; Or, (41) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 30, CDR-H2 comprising the sequence shown in SEQ ID NO: 70, and CDR-H3 comprising the sequence shown in SEQ ID NO: 113; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 153, CDR-L2 comprising the sequence shown in SEQ ID NO: 186, and CDR-L3 comprising the sequence shown in SEQ ID NO: 228; Or, (42) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 31, CDR-H2 comprising the sequence shown in SEQ ID NO: 71, and CDR-H3 comprising the sequence shown in SEQ ID NO: 114; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 154, CDR-L2 comprising the sequence shown in SEQ ID NO: 187, and CDR-L3 comprising the sequence shown in SEQ ID NO: 229; Or, (43) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 32, CDR-H2 comprising the sequence shown in SEQ ID NO: 72, and CDR-H3 comprising the sequence shown in SEQ ID NO: 115; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 155, CDR-L2 comprising the sequence shown in SEQ ID NO: 188, and CDR-L3 comprising the sequence shown in SEQ ID NO: 230; Or, (44) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 33, CDR-H2 comprising the sequence shown in SEQ ID NO: 73, and CDR-H3 comprising the sequence shown in SEQ ID NO: 116; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 156, CDR-L2 comprising the sequence shown in SEQ ID NO: 189, and CDR-L3 comprising the sequence shown in SEQ ID NO: 231; Or, (45) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 34, CDR-H2 comprising the sequence shown in SEQ ID NO: 74, and CDR-H3 comprising the sequence shown in SEQ ID NO: 117; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 157, CDR-L2 comprising the sequence shown in SEQ ID NO: 190, and CDR-L3 comprising the sequence shown in SEQ ID NO: 232; Or, (46) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 35, CDR-H2 comprising the sequence shown in SEQ ID NO: 75, and CDR-H3 comprising the sequence shown in SEQ ID NO: 118; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 158, CDR-L2 comprising the sequence shown in SEQ ID NO: 191, and CDR-L3 comprising the sequence shown in SEQ ID NO: 233; Or, (47) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 26, CDR-H2 comprising the sequence shown in SEQ ID NO: 76, and CDR-H3 comprising the sequence shown in SEQ ID NO: 119; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 159, CDR-L2 comprising the sequence shown in SEQ ID NO: 192, and CDR-L3 comprising the sequence shown in SEQ ID NO: 234; Or, (48) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 24, CDR-H2 comprising the sequence shown in SEQ ID NO: 77, and CDR-H3 comprising the sequence shown in SEQ ID NO: 120; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 160, CDR-L2 comprising the sequence shown in SEQ ID NO: 193, and CDR-L3 comprising the sequence shown in SEQ ID NO: 235; Or, (49) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 36, CDR-H2 comprising the sequence shown in SEQ ID NO: 78, and CDR-H3 comprising the sequence shown in SEQ ID NO: 121; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 161, CDR-L2 comprising the sequence shown in SEQ ID NO: 194, and CDR-L3 comprising the sequence shown in SEQ ID NO: 236; Or, (50) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 32, CDR-H2 comprising the sequence shown in SEQ ID NO: 79, and CDR-H3 comprising the sequence shown in SEQ ID NO: 122; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 162, CDR-L2 comprising the sequence shown in SEQ ID NO: 195, and CDR-L3 comprising the sequence shown in SEQ ID NO: 237; Or, (51) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 37, CDR-H2 comprising the sequence shown in SEQ ID NO: 79, and CDR-H3 comprising the sequence shown in SEQ ID NO: 123; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 163; CDR-L2 comprising the sequence shown in SEQ ID NO: 196; and CDR-L3 comprising the sequence shown in SEQ ID NO: 238; The antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL) as described above, where the CDRs are defined according to the IMGT numbering system.

[0009] In some embodiments of the present invention, (1) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 392, CDR-H2 comprising the sequence shown in SEQ ID NO: 393, and CDR-H3 comprising the sequence shown in SEQ ID NO: 394; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 395, CDR-L2 comprising the sequence shown in SEQ ID NO: 396, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; or (2) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence set forth in SEQ ID NO: 377, CDR-H2 comprising the sequence set forth in SEQ ID NO: 378, and CDR-H3 comprising the sequence set forth in SEQ ID NO: 379; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 380, CDR-L2 comprising the sequence shown in SEQ ID NO: 381, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; or (3) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 362, CDR-H2 comprising the sequence shown in SEQ ID NO: 363, and CDR-H3 comprising the sequence shown in SEQ ID NO: 364; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 365; CDR-L2 comprising the sequence shown in SEQ ID NO: 366; and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; The CDRs may be defined by the Kabat numbering system.

[0010] In some embodiments of the present invention, (1) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 397, CDR-H2 comprising the sequence shown in SEQ ID NO: 398, and CDR-H3 comprising the sequence shown in SEQ ID NO: 394; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 395, CDR-L2 comprising the sequence shown in SEQ ID NO: 396, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 382, CDR-H2 comprising the sequence shown in SEQ ID NO: 383, and CDR-H3 comprising the sequence shown in SEQ ID NO: 379; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 380, CDR-L2 comprising the sequence shown in SEQ ID NO: 381, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; Or, (3) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 367, CDR-H2 comprising the sequence shown in SEQ ID NO: 368, and CDR-H3 comprising the sequence shown in SEQ ID NO: 364; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 365; CDR-L2 comprising the sequence shown in SEQ ID NO: 366; and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; The CDRs may be numbered according to the AbM numbering system definition.

[0011] In some embodiments of the present invention, (1) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence set forth in SEQ ID NO: 399, CDR-H2 comprising the sequence set forth in SEQ ID NO: 400, and CDR-H3 comprising the sequence set forth in SEQ ID NO: 394; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 395, CDR-L2 comprising the sequence shown in SEQ ID NO: 396, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence set forth in SEQ ID NO: 384; CDR-H2 comprising the sequence set forth in SEQ ID NO: 385; and CDR-H3 comprising the sequence set forth in SEQ ID NO: 379; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 380, CDR-L2 comprising the sequence shown in SEQ ID NO: 381, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; (3) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence set forth in SEQ ID NO: 369, CDR-H2 comprising the sequence set forth in SEQ ID NO: 370, and CDR-H3 comprising the sequence set forth in SEQ ID NO: 364; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 365; CDR-L2 comprising the sequence shown in SEQ ID NO: 366; and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; The CDRs may be defined by the Chothia numbering system.

[0012] In the present invention, (1) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 401, CDR-H2 comprising the sequence shown in SEQ ID NO: 402, and CDR-H3 comprising the sequence shown in SEQ ID NO: 403; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 404, CDR-L2 comprising the sequence shown in SEQ ID NO: 405, and CDR-L3 comprising the sequence shown in SEQ ID NO: 406; Or, (2) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 386, CDR-H2 comprising the sequence shown in SEQ ID NO: 387, and CDR-H3 comprising the sequence shown in SEQ ID NO: 388; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 389, CDR-L2 comprising the sequence shown in SEQ ID NO: 390, and CDR-L3 comprising the sequence shown in SEQ ID NO: 391; Or, (3) A heavy chain variable region (VH) comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 371, CDR-H2 comprising the sequence shown in SEQ ID NO: 372, and CDR-H3 comprising the sequence shown in SEQ ID NO: 373; and A light chain variable region (VL) comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 374; CDR-L2 comprising the sequence shown in SEQ ID NO: 375; and CDR-L3 comprising the sequence shown in SEQ ID NO: 376; The CDRs may be defined by a Contact numbering system.

[0013] In some preferred embodiments, the antibodies or antigen-binding fragments thereof of the invention (1) VH of the sequence shown in SEQ ID NO: 239 and VL of the sequence shown in SEQ ID NO: 289 (2) VH of the sequence shown in SEQ ID NO: 239 and VL of the sequence shown in SEQ ID NO: 290 (3) VH of the sequence shown in SEQ ID NO: 240 and VL of the sequence shown in SEQ ID NO: 291 (4) VH of the sequence shown in SEQ ID NO: 241 and VL of the sequence shown in SEQ ID NO: 292 (5) VH of the sequence shown in SEQ ID NO: 242 and VL of the sequence shown in SEQ ID NO: 293 (6) VH of the sequence shown in SEQ ID NO: 243 and VL of the sequence shown in SEQ ID NO: 294 (7) VH of the sequence shown in SEQ ID NO: 244 and VL of the sequence shown in SEQ ID NO: 295 (8) VH of the sequence shown in SEQ ID NO: 245 and VL of the sequence shown in SEQ ID NO: 296 (9) VH of the sequence shown in SEQ ID NO: 246 and VL of the sequence shown in SEQ ID NO: 297 (10) VH of the sequence shown in SEQ ID NO: 247 and VL of the sequence shown in SEQ ID NO: 298 (11) VH of the sequence shown in SEQ ID NO: 248 and VL of the sequence shown in SEQ ID NO: 296 (12) VH of the sequence shown in SEQ ID NO: 249 and VL of the sequence shown in SEQ ID NO: 299 (13) VH of the sequence shown in SEQ ID NO: 250 and VL of the sequence shown in SEQ ID NO: 300 (14) VH of the sequence shown in SEQ ID NO: 251 and VL of the sequence shown in SEQ ID NO: 301 (15) VH of the sequence shown in SEQ ID NO: 252 and VL of the sequence shown in SEQ ID NO: 302 (16) VH of the sequence shown in SEQ ID NO: 253 and VL of the sequence shown in SEQ ID NO: 303 (17) VH of the sequence shown in SEQ ID NO: 254 and VL of the sequence shown in SEQ ID NO: 304 (18) VH of the sequence shown in SEQ ID NO: 255 and VL of the sequence shown in SEQ ID NO: 305 (19) VH of the sequence shown in SEQ ID NO: 256 and VL of the sequence shown in SEQ ID NO: 306 (20) VH of the sequence shown in SEQ ID NO: 257 and VL of the sequence shown in SEQ ID NO: 307 (21) VH of the sequence shown in SEQ ID NO: 258 and VL of the sequence shown in SEQ ID NO: 308 (22) VH of the sequence shown in SEQ ID NO: 259 and VL of the sequence shown in SEQ ID NO: 309 (23) VH of the sequence shown in SEQ ID NO: 260 and VL of the sequence shown in SEQ ID NO: 310 (24) VH of the sequence shown in SEQ ID NO: 261 and VL of the sequence shown in SEQ ID NO: 311 (25) VH of the sequence shown in SEQ ID NO: 262 and VL of the sequence shown in SEQ ID NO: 312 (26) VH of the sequence shown in SEQ ID NO: 263 and VL of the sequence shown in SEQ ID NO: 313 (27) VH of the sequence shown in SEQ ID NO: 264 and VL of the sequence shown in SEQ ID NO: 314 (28) VH of the sequence shown in SEQ ID NO: 265 and VL of the sequence shown in SEQ ID NO: 315 (29) VH of the sequence shown in SEQ ID NO: 266 and VL of the sequence shown in SEQ ID NO: 316 (30) VH of the sequence shown in SEQ ID NO: 267 and VL of the sequence shown in SEQ ID NO: 317 (31) VH of the sequence shown in SEQ ID NO: 268 and VL of the sequence shown in SEQ ID NO: 318 (32) VH of the sequence shown in SEQ ID NO: 269 and VL of the sequence shown in SEQ ID NO: 319 (33) VH of the sequence shown in SEQ ID NO: 270 and VL of the sequence shown in SEQ ID NO: 320 (34) VH of the sequence shown in SEQ ID NO: 271 and VL of the sequence shown in SEQ ID NO: 321 (35) VH of the sequence shown in SEQ ID NO: 272 and VL of the sequence shown in SEQ ID NO: 322 (36) VH of the sequence shown in SEQ ID NO: 273 and VL of the sequence shown in SEQ ID NO: 323 (37) VH of the sequence shown in SEQ ID NO: 274 and VL of the sequence shown in SEQ ID NO: 324 (38) VH of the sequence shown in SEQ ID NO: 275 and VL of the sequence shown in SEQ ID NO: 325 (39) VH of the sequence shown in SEQ ID NO: 276 and VL of the sequence shown in SEQ ID NO: 326 (40) VH of the sequence shown in SEQ ID NO: 277 and VL of the sequence shown in SEQ ID NO: 327 (41) VH of the sequence shown in SEQ ID NO: 278 and VL of the sequence shown in SEQ ID NO: 328 (42) VH of the sequence shown in SEQ ID NO: 279 and VL of the sequence shown in SEQ ID NO: 329 (43) VH of the sequence shown in SEQ ID NO: 280 and VL of the sequence shown in SEQ ID NO: 330 (44) VH of the sequence shown in SEQ ID NO: 281 and VL of the sequence shown in SEQ ID NO: 331 (45) VH of the sequence shown in SEQ ID NO: 282 and VL of the sequence shown in SEQ ID NO: 332 (46) VH of the sequence shown in SEQ ID NO: 283 and VL of the sequence shown in SEQ ID NO: 333 (47) VH of the sequence shown in SEQ ID NO: 284 and VL of the sequence shown in SEQ ID NO: 334 (48) VH of the sequence shown in SEQ ID NO: 285 and VL of the sequence shown in SEQ ID NO: 335 (49) VH of the sequence shown in SEQ ID NO: 286 and VL of the sequence shown in SEQ ID NO: 336 (50) VH of the sequence shown in SEQ ID NO: 287 and VL of the sequence shown in SEQ ID NO: 337 (51) VH of the sequence shown in SEQ ID NO: 288 and VL of the sequence shown in SEQ ID NO: 338 (52) VH of the sequence shown in SEQ ID NO: 339 and VL of the sequence shown in SEQ ID NO: 351 (53) VH of the sequence shown in SEQ ID NO: 339 and VL of the sequence shown in SEQ ID NO: 352 (54) VH of the sequence shown in SEQ ID NO: 339 and VL of the sequence shown in SEQ ID NO: 353 (55) VH of the sequence shown in SEQ ID NO: 340 and VL of the sequence shown in SEQ ID NO: 351 (56) VH of the sequence shown in SEQ ID NO: 340 and VL of the sequence shown in SEQ ID NO: 352 (57) VH of the sequence shown in SEQ ID NO: 340 and VL of the sequence shown in SEQ ID NO: 353 (58) VH of the sequence shown in SEQ ID NO: 341 and VL of the sequence shown in SEQ ID NO: 351 (59) VH of the sequence shown in SEQ ID NO: 341 and VL of the sequence shown in SEQ ID NO: 352 (60) VH of the sequence shown in SEQ ID NO: 341 and VL of the sequence shown in SEQ ID NO: 353 (61) VH of the sequence shown in SEQ ID NO: 342 and VL of the sequence shown in SEQ ID NO: 351 (62) VH of the sequence shown in SEQ ID NO: 342 and VL of the sequence shown in SEQ ID NO: 352 (63) VH of the sequence shown in SEQ ID NO: 342 and VL of the sequence shown in SEQ ID NO: 353 (64) VH of the sequence shown in SEQ ID NO: 343 and VL of the sequence shown in SEQ ID NO: 354 (65) VH of the sequence shown in SEQ ID NO: 343 and VL of the sequence shown in SEQ ID NO: 355 (66) VH of the sequence shown in SEQ ID NO: 343 and VL of the sequence shown in SEQ ID NO: 356 (67) VH of the sequence shown in SEQ ID NO: 344 and VL of the sequence shown in SEQ ID NO: 354 (68) VH of the sequence shown in SEQ ID NO: 344 and VL of the sequence shown in SEQ ID NO: 355 (69) VH of the sequence shown in SEQ ID NO: 344 and VL of the sequence shown in SEQ ID NO: 356 (70) VH of the sequence shown in SEQ ID NO: 345 and VL of the sequence shown in SEQ ID NO: 354 (71) VH of the sequence shown in SEQ ID NO: 345 and VL of the sequence shown in SEQ ID NO: 355 (72) VH of the sequence shown in SEQ ID NO: 345 and VL of the sequence shown in SEQ ID NO: 356 (73) VH of the sequence shown in SEQ ID NO: 346 and VL of the sequence shown in SEQ ID NO: 354 (74) VH of the sequence shown in SEQ ID NO: 346 and VL of the sequence shown in SEQ ID NO: 355 (75) VH of the sequence shown in SEQ ID NO: 346 and VL of the sequence shown in SEQ ID NO: 356 (76) VH of the sequence shown in SEQ ID NO: 347 and VL of the sequence shown in SEQ ID NO: 357 (77) VH of the sequence shown in SEQ ID NO: 347 and VL of the sequence shown in SEQ ID NO: 358 (78) VH of the sequence shown in SEQ ID NO: 347 and VL of the sequence shown in SEQ ID NO: 359 (79) VH of the sequence shown in SEQ ID NO: 348 and VL of the sequence shown in SEQ ID NO: 357 (80) VH of the sequence shown in SEQ ID NO: 348 and VL of the sequence shown in SEQ ID NO: 358 (81) VH of the sequence shown in SEQ ID NO: 348 and VL of the sequence shown in SEQ ID NO: 359 (82) VH of the sequence shown in SEQ ID NO: 349 and VL of the sequence shown in SEQ ID NO: 357 (83) VH of the sequence shown in SEQ ID NO: 349 and VL of the sequence shown in SEQ ID NO: 358 (84) VH of the sequence shown in SEQ ID NO: 349 and VL of the sequence shown in SEQ ID NO: 359 (85) VH of the sequence shown in SEQ ID NO: 350 and VL of the sequence shown in SEQ ID NO: 357 (86) VH of the sequence shown in SEQ ID NO: 350 and VL of the sequence shown in SEQ ID NO: 358 (87) VH of the sequence shown in SEQ ID NO: 350 and VL of the sequence shown in SEQ ID NO: 359 It includes any combination selected from the above heavy chain variable region (VH) and light chain variable region (VL).

[0014] Optionally, the heavy chain variable region (VH) and the light chain variable region (VL) have at least 70%, 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%, at least 99%, or 100% sequence identity to any combination of (1) to (87), and / or the light chain variable region (VL) has at least 70%, 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%, at least 99%, or 100% sequence identity.

[0015] In some preferred embodiments, the antibody or antigen-binding fragment thereof of the present invention is selected from a Fab fragment, a Fab' fragment, a F(ab)'2 fragment, a single-chain antibody, and a disulfide-stabilized Fv protein (dsFv).

[0016] In some preferred embodiments, the antibodies or antigen-binding fragments thereof of the invention are single chain antibodies, such as scFv, di-scFv or (scFv)2.

[0017] In some preferred embodiments, in the antibodies or antigen-binding fragments thereof of the present invention, the antibodies or antigen-binding fragments thereof are murine antibodies, chimeric antibodies, humanized antibodies, or antibodies derived from other species (e.g., rabbit, camel, or shark).

[0018] In some preferred embodiments, the antibodies or antigen-binding fragments thereof of the present invention further comprise a heavy chain constant region (CH) and a light chain constant region (CL).

[0019] In some preferred embodiments, the heavy chain constant region is selected from the heavy chain constant regions of IgG, IgM, IgE, IgD, and IgA, or variants thereof.

[0020] In some preferred embodiments, the light chain constant region is selected from a kappa or lambda light chain constant region or variant thereof.

[0021] In some preferred embodiments, the variant has one or more amino acid substitutions, deletions or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions or additions) relative to the wild-type sequence from which it is derived.

[0022] In some preferred embodiments, the antibodies or antigen-binding fragments thereof of the invention (1) Human IgG1 heavy chain constant region (2) Human IgG1 light chain constant region It includes a heavy chain constant region and a light chain constant region selected from the above.

[0023] In some preferred embodiments, the heavy chain constant region (CH) of an antibody or antigen-binding fragment thereof of the invention comprises the amino acid sequence set forth in SEQ ID NO:360.

[0024] In some preferred embodiments, the light chain constant region is a human IgG1 κ light chain constant region, and the light chain constant region (CL) comprises the amino acid sequence set forth in SEQ ID NO:361.

[0025] In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises a light chain constant region (CL) set forth in SEQ ID NO: 361 or a variant thereof, wherein the variant has up to 20 amino acid conservative substitutions (e.g., up to 15, up to 10, or up to 5 amino acid conservative substitutions, e.g., 1, 2, 3, 4, or 5 amino acid conservative substitutions) relative to SEQ ID NO: 361.

[0026] In some preferred embodiments, the antibodies or antigen-binding fragments thereof of the invention have a K of less than about 500 nM, e.g., less than about 100 nM, less than about 50 nM, less than about 40 nM, less than about 40 nM, less than about 20 nM, less than about 10 nM, less than about 1 nM, less than about 0.1 nM, less than about 0.01 nM or lower. D It binds to CDCP1 (e.g., human CDCP1) at

[0027] In some preferred embodiments, the antibodies or antigen-binding fragments thereof of the invention have an EC50 of less than about 500 nM, e.g., less than about 100 nM, less than about 10 nM, less than about 1 nM, less than about 0.9 nM, less than about 0.8 nM, less than about 0.7 nM, less than about 0.6 nM, less than about 0.5 nM, less than about 0.4 nM, less than about 0.3 nM, less than about 0.2 nM, less than about 0.1 nM, less than about 0.01 nM or less. 50 and binds to tumor cells expressing CDCP1, preferably the EC 50 is measured by flow cytometry or competitive ELISA.

[0028] In some preferred embodiments, the antibody or antigen-binding fragment thereof does not have or has endocytic capabilities.

[0029] In some preferred embodiments, the antibody or antigen-binding fragment thereof does not have the ability to promote tumor cell migration.

[0030] In some preferred embodiments, the antibody or antigen-binding fragment thereof has the ability to inhibit tumor metastasis.

[0031] (induced antibody) The antibodies or antigen-binding fragments thereof of the present invention may be derivatized, e.g., linked to another molecule (e.g., another polypeptide or protein). Generally, derivatization (e.g., labeling) of an antibody or antigen-binding fragment thereof does not adversely affect its binding to CDCP1 (particularly human CDCP1). Thus, the antibodies or antigen-binding fragments thereof of the present invention are also intended to include such derivatized forms. For example, the antibodies or antigen-binding fragments thereof of the present invention may be functionally linked (by chemical conjugation, genetic fusion, non-covalent bonding, or other means) to one or more other molecular groups, e.g., another antibody (e.g., to form a bispecific antibody), a detection reagent, a medicinal reagent, and / or a protein or polypeptide that can mediate binding of the antibody or antigen-binding fragment to another molecule (e.g., avidin or a polyhistidine tag).

[0032] One type of derivatized antibody (e.g., bispecific antibodies) is produced by crosslinking two or more antibodies (of the same type or different types). Methods for obtaining bispecific antibodies are well known in the art and include, but are not limited to, chemical crosslinking, cell engineering (hybridoma) or genetic engineering techniques.

[0033] Another type of derivatized antibody is an antibody linked to a therapeutic moiety. The therapeutic moiety according to the present invention may be a bacterial toxin, a cytotoxic drug, or a radiotoxin, including, but not limited to, paclitaxel, cytochalasin B, mitomycin, etoposide, vincristine, or other antimetabolites, alkylating agents, antibiotics, or antimitotics.

[0034] Another type of derivatized antibody is a labeled antibody. For example, an antibody of the present invention or an antigen-binding fragment thereof may be linked to a detectable label. A detectable label according to the present invention may be any substance that is detectable by fluorescent, spectroscopic, photochemical, biochemical, immunological, electrical, optical, or chemical means. Such labels are well known in the art and include enzymes (e.g., horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radioisotopes (e.g., 3 H, 125 I, 35 S, 14 C or 32Examples of suitable substrates include, but are not limited to, 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)), acridinium ester compounds, magnetic beads, calorimetric markers (e.g., gold colloids, colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.)), and biotin for binding avidin (e.g., streptavidin) modified with the marker. Patents teaching the use of such markers include, but are not limited to, U.S. Patent Nos. 3,817,837, 3,850,752, 3,939,350, 3,996,345, 4,277,437, 4,275,149, and 4,366,241 (incorporated herein by reference in their entirety). The detectable labels may be detected by methods known in the art. For example, radioactive labels may be detected with photographic film or a scintillation counter, and fluorescent markers may be detected by detecting emitted light with a photodetector. Enzymatic markers are generally detected by providing the enzyme with a substrate and detecting the reaction product produced by the action of the enzyme on the substrate, and calorimetric markers are detected with an easily visualized color marker. In some embodiments, such labels are suitable for immunological detection (e.g., enzyme-linked immunosorbent assay, radioimmunoassay, fluorescent immunoassay, chemiluminescent immunoassay, etc.). In some embodiments, the detectable label may be linked to the antibody or antigen-binding fragment thereof of the invention by linkers of different lengths to reduce potential steric hindrance.

[0035] The antibodies of the invention or antigen-binding fragments thereof may also be derivatized with chemical groups, such as polyethylene glycol (PEG), methyl or ethyl groups, or glycosyl groups, which are used to improve the biological properties of the antibody, e.g., to increase serum half-life.

[0036] (multispecific antibody) In another aspect of the invention, multispecific antibodies are provided, the multispecific antibodies comprising an antibody of the invention or an antigen-binding fragment thereof, and in some preferred embodiments the multispecific antibody is a bispecific antibody, a trispecific antibody or a tetraspecific antibody.

[0037] (isolated nucleic acid molecule) In another aspect of the invention, there is provided a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof of the invention, or a multispecific antibody of the invention.

[0038] (vector) Another aspect of the invention provides a vector comprising a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof of the invention, or a multispecific antibody of the invention.

[0039] In some preferred embodiments, the vector is a cloning vector or an expression vector.

[0040] In some preferred embodiments, the vectors of the invention are, for example, plasmids, cosmids, phages, etc. In some preferred embodiments, the vectors are capable of expressing an antibody or antigen-binding fragment thereof of the invention in a subject (e.g., a mammal (e.g., a human)).

[0041] (host cell) Another aspect of the present invention provides host cells comprising the nucleic acid molecules of the present invention or the vectors of the present invention. Such host cells include, but are not limited to, prokaryotic cells (e.g., E. coli cells) and eukaryotic cells (e.g., yeast cells, insect cells, plant cells, and animal cells (e.g., mammalian cells, e.g., mouse cells, human cells, etc.)). In some preferred embodiments, the host cells of the present invention are mammalian cells, such as CHO cells (e.g., CHO-K1, CHO-S, CHO DG44).

[0042] (antibody production) In another aspect, the present invention provides a method for producing an antibody or antigen-binding fragment thereof, or a multispecific antibody that specifically binds to CDCP1, comprising culturing a host cell of the present invention under conditions allowing expression of the antibody or antigen-binding fragment thereof, or the multispecific antibody, and recovering the antibody or antigen-binding fragment thereof, or the multispecific antibody from the cultured host cell culture.

[0043] The antibodies of the present invention may be produced by various methods known in the art, for example, by recombinant genetic engineering techniques. For example, DNA molecules encoding the heavy and light chain genes of the antibodies of the present invention are obtained by chemical synthesis or PCR amplification. The obtained DNA molecules are inserted into an expression vector, which is then transfected into host cells. The transfected host cells are then cultured under specific conditions to express the antibodies of the present invention.

[0044] Antigen-binding fragments of the present invention may be obtained by hydrolysis of intact antibody molecules (see Morimoto et al., J. Biochem. Biophys. Methods 24:107-117 (1992); Brennan et al., Science 229:81 (1985)). Alternatively, these antigen-binding fragments may be produced directly from recombinant host cells (see Hudson, Curr. Opin. Immunol. 11:548-557 (1999); Little et al., Immunol. Today, 21:364-370 (2000)). For example, Fab' fragments may be obtained directly from host cells and chemically coupled to form F(ab')2 fragments (Carter et al., Bio / Technology, 10:163-167 (1992)). Alternatively, Fv, Fab or F(ab')2 fragments may be isolated directly from recombinant host cell culture, and those skilled in the art will be familiar with other techniques for producing such antigen-binding fragments.

[0045] (complex) In another aspect of the invention, there is provided a conjugate, or a pharmaceutically acceptable salt thereof, comprising an antibody or antigen-binding fragment thereof, or a multispecific antibody of the invention, and a conjugate moiety, wherein the antibody or antigen-binding fragment thereof, or the multispecific antibody is linked to the conjugate moiety directly or via a linker. In some preferred embodiments, the conjugate moiety is selected from a detectable label (e.g., a radioisotope, a fluorescent substance, a luminescent substance, a colored substance, or an enzyme) or a therapeutic agent (e.g., a nuclide, an immunostimulatory agent, an immunosuppressant, a cytokine, a toxin, and other active agents that inhibit tumor cell growth or promote tumor cell apoptosis or necrosis).

[0046] In some preferred embodiments, the structure of the complex is shown in formula (I): A-(LD) p (I) with or without L, wherein A is an antibody or antigen-binding fragment thereof of the present invention, or a multispecific antibody according to the present invention; L is a linker, preferably a vc linker having the structure: [ka] and D is a conjugated moiety.

[0047] In some preferred embodiments, the conjugate moiety is an immunostimulant, including a TLR, STING, OND, CpG, LPS, SEB, SEA, or SEC stimulant.

[0048] In some preferred embodiments, the conjugate moiety is a cytokine and includes IL-6, IL-15, IL-12, IL-1b, or IL-23.

[0049] In some preferred embodiments, the conjugate moiety is a toxin and comprises a cytotoxic agent, more preferably the cytotoxic agent is selected from camptothecins (e.g., SN-38), maytansinoids (e.g., maytansinoid DM1 / 4), pyrrolobenzodiazepines (PBDs), or auristatins (e.g., monomethyl auristatin E / F).

[0050] In some preferred embodiments, the value of p is 0 or greater.

[0051] In some preferred embodiments, p is 0-8.

[0052] (Pharmaceutical composition) In another aspect of the invention, there is provided a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof, or a prodrug thereof (e.g., a probody), conjugate, or multispecific antibody of the invention, and a pharmaceutically acceptable carrier and / or excipient.

[0053] In some preferred embodiments, the pharmaceutical composition further comprises another pharmaceutically active agent. In some preferred embodiments, the other pharmaceutically active agent is a drug with anti-tumor activity, such as a platinum-based drug or a small molecule inhibitor. In some preferred embodiments, the other pharmaceutically active agent is an immune enhancing agent, such as an interferon or an interleukin. In some preferred embodiments, the other pharmaceutically active agent is an immunotherapeutic agent, such as an immune checkpoint inhibitor (e.g., a PD-1, PD-L1, or CTLA-4 inhibitor), an immune enhancing agent (e.g., an interferon or an interleukin), or an immunostimulator (e.g., a TLR, OND, CpG, LPS, SEB, SEA, STING, or SEC activator). In some preferred embodiments, the other pharmaceutically active agent is an oncolytic virus, an immune cell, or an engineered immune cell. In some preferred embodiments, the other pharmaceutically active agent is a radioactive substance or a color-producing agent.

[0054] In some preferred embodiments, the antibody or antigen-binding fragment thereof and the additional pharmaceutically active agent are provided as separate components or as components of the same composition.

[0055] In some preferred embodiments, the antibody or antigen-binding fragment thereof, conjugate, multispecific antibody, or pharmaceutical composition of the present invention is used to (a) induce apoptosis of tumor cells, (b) suppress tumor cell proliferation, (c) induce and / or increase T cell infiltration, (d) induce and / or enhance an immune response, (e) induce and / or enhance complement-dependent cytotoxicity, (f) induce and / or enhance antibody-dependent cellular cytotoxicity, (g) enhance NK cell activity, (h) suppress CDCP1 expression and activation, (i) suppress CDCP1-mediated signaling, or (j) any combination of (a)-(i) in a subject.

[0056] In some preferred embodiments, the tumor is selected from a solid tumor or a hematological tumor.

[0057] In some preferred embodiments, the tumor is a CDCP1-positive tumor.

[0058] In some preferred embodiments, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, and glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

[0059] (Application) In another aspect of the present invention, there is provided use of the antibody or antigen-binding fragment thereof, multispecific antibody, conjugate, or pharmaceutical composition of the present invention for producing a medicament for the prevention and / or treatment and / or adjuvant therapy and / or neoadjuvant therapy of a CDCP1-associated disease.

[0060] In some preferred embodiments, the CDCP1-associated disease is a tumor.

[0061] In some preferred embodiments, the tumor is selected from a solid tumor or a hematological tumor.

[0062] In some preferred embodiments, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

[0063] In some preferred embodiments, the antibody or antigen-binding fragment thereof, conjugate, multispecific antibody or pharmaceutical composition of the invention is administered separately, in combination, simultaneously or sequentially with another pharmaceutically active agent.

[0064] In some preferred embodiments, the additional pharmaceutically active agent is a drug with anti-tumor activity, such as a platinum-based drug, a small molecule inhibitor, or the like.

[0065] In some preferred embodiments, the additional pharmaceutically active agent is an immune enhancing agent, eg, an interferon or an interleukin.

[0066] In some preferred embodiments, the additional pharmaceutically active agent is an immunostimulant (e.g., a TLR, OND, CpG, LPS, SEB, STING, SEA, or SEC stimulant).

[0067] In some preferred embodiments, the additional pharmaceutically active agent is an immune checkpoint inhibitor (e.g., a PD-1, PD-L1, or CTLA-4 inhibitor).

[0068] In some preferred embodiments, the additional pharmaceutically active agent is an oncolytic virus, an immune cell, or an engineered immune cell.

[0069] In another aspect of the present invention, there is provided a method for the prevention and / or treatment and / or adjuvant therapy and / or neoadjuvant therapy of a CDCP1-associated disease in a subject, said method comprising administering to a subject in need thereof an effective amount of an antibody or antigen-binding fragment thereof, conjugate, multispecific antibody, or pharmaceutical composition of the present invention.

[0070] In some preferred embodiments, the method further comprises administering to the subject a second therapy, wherein the second therapy is selected from surgery, chemotherapy, radiation therapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, viral therapy, adjuvant therapy, cell therapy, and any combination thereof, and wherein the second therapy may be used separately or in combination with the method of the present invention, and wherein the second therapy may be used separately or in combination with the method of the present invention, simultaneously or sequentially.

[0071] In some preferred embodiments, the CDCP1-associated disease is a tumor.

[0072] In some preferred embodiments, the tumor is selected from a solid tumor or a hematological tumor.

[0073] In some preferred embodiments, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

[0074] In another aspect of the present invention, there is provided a method for detecting the presence or level of CDCP1 in a sample, comprising contacting the sample with the antibody or antigen-binding fragment thereof, multispecific antibody, conjugate, or pharmaceutical composition of the present invention under conditions that allow the antibody or antigen-binding fragment thereof, multispecific antibody, conjugate, or pharmaceutical composition to form a complex with CDCP1, and detecting the formation of the complex.

[0075] In another aspect of the present invention, there is provided an antibody or an antigen-binding fragment thereof, a conjugate, a multispecific antibody, or a pharmaceutical composition of the present invention for use in the prevention and / or treatment and / or adjuvant therapy and / or neoadjuvant therapy of a CDCP1-associated disease.

[0076] In some preferred embodiments, the CDCP1-associated disease is a tumor.

[0077] In some preferred embodiments, the tumor is selected from a solid tumor or a hematological tumor.

[0078] In some preferred embodiments, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

[0079] In another aspect of the present invention, there is provided an antibody or antigen-binding fragment thereof, conjugate, multispecific antibody or pharmaceutical composition of the present invention for use in the diagnosis or differential diagnosis of a CDCP1-associated disease.

[0080] In some preferred embodiments, the CDCP1-associated disease is a tumor.

[0081] In some preferred embodiments, the tumor is selected from a solid tumor or a hematological tumor.

[0082] In some preferred embodiments, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

[0083] The antibody or antigen-binding fragment thereof, pharmaceutical composition, or immunogenic composition of the present invention may be formulated into any dosage form known in the medical field, such as tablets, pills, suspensions, emulsions, solutions, gelling agents, capsules, powders, granules, elixirs, tablets, suppositories, injections (including injection solutions, sterile powders for injection, and concentrated solutions for injection), inhalants, sprays, etc. The preferred dosage form is determined by the desired mode of administration and therapeutic application. The pharmaceutical compositions of the present invention are sterile and stable under the conditions for production and storage. One preferred dosage form is an injection. Such an injection may be a sterile injection solution. For example, a sterile injection solution may be prepared by the following method: A predetermined dose of the recombinant protein of the present invention is added to an appropriate solvent, and optionally, other desired ingredients (including, but not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, tonicity adjusters, preservatives, diluents, or any combination thereof) are added simultaneously, followed by filtration and sterilization. For ease of storage and use, the sterile injection solutions may also be made into sterile lyophilized powders (e.g., by vacuum drying or freeze-drying), which may be dispersed in a suitable carrier, e.g., sterile pyrogen-free water, prior to use.

[0084] The antibodies or antigen-binding fragments thereof of the present invention may also be present in pharmaceutical or immunogenic compositions in unit dosage form for ease of administration.

[0085] The antibody or antigen-binding fragment thereof, pharmaceutical composition, or immunogenic composition of the present invention may be administered by any suitable method known in the art, including, but not limited to, orally, bucally, sublingually, intraocularly, topically, parenterally, rectally, intramedullary, intracytoplasmic reticulum, inguinal, intravesically, topically (such as powders, plasters, or drops), or intranasally. However, for many therapeutic applications, parenteral administration (e.g., intravenous injection, subcutaneous injection, intraperitoneal injection, intramuscular injection) is the preferred route / mode of administration. It will be appreciated by those of skill in the art that the route and / or mode of administration will vary depending on the intended purpose. In a preferred embodiment, the antibody or antigen-binding fragment thereof, pharmaceutical composition, or immunogenic composition of the present invention is administered by intravenous infusion or injection.

[0086] The pharmaceutical or immunogenic compositions of the invention may comprise an "effective amount" ("therapeutically effective amount" or "prophylactically effective amount") of an antibody or antigen-binding fragment thereof of the invention. A "prophylactically effective amount" refers to an amount sufficient to prevent, prevent, or delay the onset of disease. A "therapeutically effective amount" refers to an amount sufficient to cure or at least partially prevent the disease and its complications in a patient suffering from the disease. The therapeutically effective amount of an antibody or antigen-binding fragment thereof of the invention will vary depending on factors such as the severity of the disease being treated, the overall state of the patient's own immune system, the patient's general condition (e.g., age, weight, sex), the mode of drug administration, and other concurrently administered treatments.

[0087] In the present invention, the dosage regimen may be adjusted to obtain the best desired response (e.g., a therapeutic or prophylactic response), for example, by administering a single dose, multiple doses over a given period of time, or the dose may be proportionally reduced or increased depending on the exigencies of treatment.

[0088] It should be noted that the dosage will vary depending on the type and severity of the condition being treated. Furthermore, those skilled in the art will understand that for a particular patient, a specific dosing regimen should be adjusted over time according to the patient's needs and the physician's professional evaluation. The dosage ranges provided herein are illustrative and are not intended to limit the use or scope of the pharmaceutical or immunogenic compositions of the present invention.

[0089] In the present invention, the subject may be a mammal, for example a human.

[0090] The invention provides a container (e.g., a plastic or glass vial (e.g., with a cap or chromatography column, hollow needle, or syringe barrel)) containing either an antibody or antigen-binding fragment, or pharmaceutical composition of the invention. The invention also provides an injection device containing either an antibody or antigen-binding fragment, or pharmaceutical composition of the invention.

[0091] (kit) The antibody or antigen-binding fragment thereof of the present invention can specifically bind to CDCP1 and can therefore be used to detect the presence or level of CDCP1 in a sample.

[0092] Thus, in another aspect of the present invention, there is provided a kit comprising an antibody or antigen-binding fragment thereof, multispecific antibody, conjugate, or pharmaceutical composition of the present invention. In some preferred embodiments, the antibody or antigen-binding fragment thereof of the present invention comprises a detectable label. In a preferred embodiment, the kit further comprises a second antibody that specifically recognizes the antibody or antigen-binding fragment thereof of the present invention. Preferably, the second antibody further comprises a detectable label.

[0093] In the present invention, the detectable label may be any substance that can be detected by fluorescent, spectroscopic, photochemical, biochemical, immunological, electrical, optical or chemical means. Particularly preferably, such a label is suitable for immunological detection (e.g., enzyme-linked immunosorbent assay, radioimmunoassay, fluorescent immunoassay, chemiluminescent immunoassay, etc.). Such labels are well known in the art and include enzymes (e.g., horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radioisotopes (e.g., 3 H, 125 I, 35 S, 14 C 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)), acridinium ester compounds, magnetic beads, calorimetric markers (e.g., gold colloids, colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.)), and biotin for binding avidin (e.g., streptavidin) modified with the marker. Patents teaching the use of such markers include, but are not limited to, U.S. Patent Nos. 3,817,837, 3,850,752, 3,939,350, 3,996,345, 4,277,437, 4,275,149, and 4,366,241 (incorporated herein by reference in their entirety). Markers covered by the present invention may be detected by methods known in the art. For example, radioactive labels may be detected with photographic film or a scintillation counter, and fluorescent markers may be detected by detecting emitted light with a photodetector. Enzymatic markers are generally detected by providing the enzyme with a substrate and detecting the reaction product produced by the action of the enzyme on the substrate, and calorimetric markers are detected with an easily visualized color marker. In some embodiments, the detectable labels may be linked to the recombinant proteins of the present invention by linkers of different lengths to reduce potential steric hindrance.

[0094] In another aspect, the present invention provides use of the antibody or antigen-binding fragment thereof, conjugate, multispecific antibody, or pharmaceutical composition of the present invention for producing a diagnostic kit, which is used for the diagnosis or differential diagnosis of a CDCP1-associated disease.

[0095] In some preferred embodiments, the CDCP1-associated disease is a tumor.

[0096] In some preferred embodiments, the tumor is selected from a solid tumor or a hematological tumor.

[0097] In some preferred embodiments, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

[0098] (Definition of terms) In the present invention, unless otherwise specified, scientific and technical terms used herein have the common meanings understood by those skilled in the art. In addition, the laboratory virological, biochemical, and immunological procedures used herein are all routine steps commonly performed in the relevant fields. In addition, definitions and interpretations of relevant terms are provided to facilitate a better understanding of the present invention.

[0099] When the terms "for example," "such as," "such as," "includes," "including," or variations thereof are used in this specification, these terms are not to be considered as limiting terms and should be understood to mean "including but not limited to."

[0100] Unless otherwise indicated herein or clearly contradicted by context, the terms "a," "an," "one," "the," and similar demonstratives are to be understood to cover both the singular and the plural in the context of describing the invention (particularly in the claims that follow).

[0101] As used herein, the term "antibody" generally refers to an immunoglobulin molecule composed of two pairs of polypeptide chains, each having one light chain (LC) and one heavy chain (HC). Antibody light chains can be classified as kappa (κ) and lambda (λ) light chains. Heavy chains can be classified as μ, δ, γ, α, or ε, and antibody isotypes are defined as IgM, IgD, IgG, IgA, and IgE, respectively. In light and heavy chains, the variable and constant regions are connected by a "J" region consisting of about 12 or more amino acids, and heavy chains further contain a "D" region consisting of about 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). For example, the heavy chain constant region of IgG 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 domains are not directly involved in binding of antibodies to antigens, but exhibit various effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement pathway (C1q). The VH and VL regions can be further divided into highly variable regions (called complementarity-determining regions (CDRs)) interspersed with conserved regions called framework regions (FRs). Each V H and V L Each of these regions or domains consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of each pair of heavy and light chains (VH and VL) form an antigen-binding site. The amino acid assignment in each region or domain may follow the definitions in Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917, and Chothia et al. (1989) Nature 342:878-883.

[0102] As used herein, unless the context clearly dictates otherwise, when the term "antibody" is referred to it not only includes complete antibodies but also antigen-binding fragments of antibodies.

[0103] As used herein, the term "complementarity determining region" or "CDR" refers to the amino acid residues in an antibody variable region that are involved in antigen binding. Each of the heavy and light chain variable regions contains three CDRs, designated CDR1, CDR2, and CDR3. The exact boundaries of these CDRs may be defined by various numbering systems known in the art, such as the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), the Chothia numbering system (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917, Chothia et al. (1989) Nature 342:878-883), the IMGT numbering system (Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003), the AbM numbering system (Martin ACR, Cheetham JC, Rees AR (1989) Modeling antibody hypervariable loops: A combined algorithm. Proc Natl Acad Sci USA 86:9268-9272) or the Contact numbering system (MacCallum, RM, Martin, ACR, & Thornton, JM (1996). Antibody-antigen Interactions: Contact Analysis and Binding Site Topography. Journal of Molecular Biology, 262(5), 732-745.). For a given antibody, those skilled in the art can easily identify the CDRs defined by each numbering system. Furthermore, the correspondence between different numbering systems is well known to those skilled in the art (see, for example, Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).

[0104] In the present invention, the CDRs contained in the antibodies or antigen-binding fragments thereof of the present invention may be identified by various numbering systems known in the art, such as the Kabat numbering system, the Chothia numbering system, the IMGT numbering system, the AbM numbering system, or the Contact numbering system. In some embodiments, the CDRs contained in the antibodies or antigen-binding fragments thereof of the present invention are identified by the Kabat numbering system, the Chothia numbering system, the IMGT numbering system, the AbM numbering system, or the Contact numbering system.

[0105] As used herein, the term "framework region" or "FR" residues refers to amino acid residues other than the CDR residues as defined above in an antibody variable region.

[0106] The term "antibody" is not intended to be limited to any particular method of antibody production. It includes, for example, recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. The antibody may be of a different isotype, such as an IgG antibody (e.g., an IgG1, IgG2, IgG3, or IgG4 subtype), an IgA1 antibody, an IgA2 antibody, an IgD antibody, an IgE antibody, or an IgM antibody.

[0107] As used herein, the term "antigen-binding fragment of an antibody" refers to a polypeptide, including a fragment of a full-length antibody, that retains the ability to specifically bind to the same antigen bound by the full-length antibody and / or specifically binds to an antigen in competition with the full-length antibody, also referred to as an "antigen-binding portion." See generally Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd ed., Raven Press, NY (1989)), which is incorporated herein by reference in its entirety for all purposes. Antigen-binding fragments of antibodies may also be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Non-limiting examples of antigen-binding fragments include Fab, Fab', F(ab')2, Fd, Fv, complementarity-determining region (CDR) fragments, scFv, diabodies, single domain antibodies, chimeric antibodies, linear antibodies, nanobodies (based on Domantis technology), probodies, and other such polypeptides that contain at least a sufficient portion of an antibody to confer specific antigen-binding ability.

[0108] As used herein, the term "full-length antibody" refers to an antibody consisting of two "full-length heavy chains" and two "full-length light chains." Here, a "full-length heavy chain" refers to a polypeptide chain consisting of, from N- to C-terminus, a heavy chain variable region (VH), a heavy chain constant region CH1 domain, a hinge region (HR), a heavy chain constant region CH2 domain, and a heavy chain constant region CH3 domain. If the full-length antibody is an IgE isotype, it optionally further contains a heavy chain constant region CH4 domain. Preferably, a "full-length heavy chain" is a polypeptide chain consisting of, from N- to C-terminus, a VH, CH1, HR, CH2, and CH3. A "full-length light chain" is a polypeptide chain consisting, from N- to C-terminus, of a light chain variable region (VL) and a light chain constant region (CL). The two pairs of full-length antibody chains are linked by a disulfide bond between the CL and CH1 and a disulfide bond between the HRs of the two full-length heavy chains. A full-length antibody contains two antigen-binding sites, each formed by a pair of VH and VL, which specifically recognize and bind to the same antigen.

[0109] As used herein, the term "Fab fragment" refers to an antibody fragment consisting of the VL, VH, CL, and CH1 domains; the term "F(ab')2 fragment" refers to an antibody fragment comprising two Fab fragments linked by a disulfide bridge in the hinge region; and the term "Fab' fragment" refers to a fragment obtained by reducing the disulfide bond linking the two heavy chain fragments of the F(ab')2 fragment, and consists of one complete light chain and an Fd fragment of a heavy chain (consisting of the VH and CH1 domains).

[0110] As used herein, the term "Fv" refers to an antibody fragment consisting of the VL and VH domains of a single antibody arm. An Fv fragment is generally considered to be the smallest antibody fragment capable of forming a complete antigen-binding site. Generally, six CDRs are considered to confer antigen-binding specificity to an antibody. However, even a single variable region (e.g., an Fd fragment containing only three antigen-specific CDRs) can recognize and bind to an antigen, although with lower affinity than the complete binding site.

[0111] As used herein, the term "Fc domain" or "Fc region" refers to a region comprising a portion of the heavy chain constant region, CH2 and CH3. The Fc fragment of an antibody has various functions but is not involved in antigen binding. "Effector functions" mediated by the Fc region include Fc receptor binding, Clq binding and complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation. In some embodiments, the Fc region comprises a hinge, CH2, and CH3. When the Fc region comprises a hinge, the hinge regulates dimerization between two polypeptides comprising the Fc. The Fc region may be any antibody heavy chain constant region isotype, such as IgG1, IgG2, IgG3, or IgG4.

[0112] The Fc domain may comprise a native Fc region or a variant Fc region. A native Fc region comprises an amino acid sequence that corresponds to the amino acid sequence of an Fc region found in nature. For example, native-sequence human Fc regions include native-sequence human IgG1 Fc regions (non-A and A allotypes), native-sequence human IgG2 Fc regions, native-sequence human IgG3 Fc regions, native-sequence human IgG4 Fc regions, and naturally occurring variants thereof. A variant Fc region comprises an amino acid sequence that differs from the amino acid sequence of a native-sequence Fc region by at least one amino acid modification. In some embodiments, the variant Fc region may have an altered effector function (e.g., Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, or complement function) relative to the native Fc region.

[0113] As used herein, the term "scFv" refers to a single polypeptide chain comprising a VL and a VH domain, wherein the VL and VH are linked by a linker. Such scFv molecules may have the general structure NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeated GGGGS amino acid sequences or variants thereof. For example, a linker having the amino acid sequence (GGGGS)4 may be used, although variants thereof may also be used. Optionally, there may be a disulfide bond between the scFv and the VH and VL.

[0114] As used herein, the term "diabody" means a VH and VL domain expressed on a single polypeptide chain, but using a linker that is too short to prevent pairing between the two domains on the same chain, allowing the domains to pair with the complementary domains of another chain and produce two antigen-binding sites (see, e.g., Holliger P. et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993); Poljak RJ et al., Structure 2:1121-1123 (1994)).

[0115] Each of the above antibody fragments retains the ability to specifically bind to the same antigen as the full-length antibody and / or specifically binds to the antigen in competition with the full-length antibody.

[0116] As used herein, a "bispecific antibody" refers to a conjugate (complex) formed by conjugating arms of a first antibody (fragment) and a second antibody (fragment) or antibody analog, and conjugation methods include, but are not limited to, chemical reaction, gene fusion, protein fusion, polypeptide fusion, and enzymatic methods. "Multispecific antibodies" include, for example, triabodies and tetrabodies, the former being antibodies with three different antigen-binding specificities, and the latter being antibodies with four different antigen-binding specificities.

[0117] As used herein, techniques for obtaining antibodies include obtaining antigen-binding fragments of antibodies (e.g., the antibody fragments described above) from a given antibody (e.g., an antibody provided by the present invention) using conventional techniques known to those skilled in the art (e.g., recombinant DNA techniques or enzymatic or chemical cleavage methods), and screening the antigen-binding fragments of antibodies for specificity in the same manner as used for intact antibodies.

[0118] As used herein, the terms "monoclonal antibody," "mab," and "mAb" have the same meaning and may be used interchangeably to refer to an antibody or antibody fragment derived from a highly homologous series of antibody molecules that are identical absent spontaneous, natural mutations. Monoclonal antibodies have high specificity for a single epitope on an antigen. Polyclonal antibodies, in contrast to monoclonal antibodies, generally include at least two or more different antibodies that recognize different epitopes on an antigen. The qualifier "monoclonal" indicates that the antibody is characterized as being obtained from a highly homologous group of antibodies, and does not imply that the antibody is produced by a specific method.

[0119] As used herein, the term "myelin antibody" refers to a substance obtained by fusing B cells from an immunized mouse with myeloma cells, screening for mouse hybrid fusion cells capable of indefinite proliferation and antibody secretion, followed by further screening, antibody production, and antibody purification. Alternatively, an antigen enters the mouse's body, causing B cells to differentiate and proliferate to form plasma cells, which can produce secretory antibodies. Antibodies are produced in response to stimulation by a specific antigen; when an antigen enters the human body and triggers interactions between various immune cells, B cells (a type of lymphocyte) differentiate and proliferate to form plasma cells, which can produce secretory antibodies.

[0120] As used herein, the term "humanized antibody" refers to a genetically engineered non-human antibody whose amino acid sequence has been modified to enhance its homology with that of a human antibody. Generally, all or a portion of the CDR regions of a humanized antibody are derived from a non-human antibody (donor antibody), and all or a portion of the non-CDR regions (e.g., variable region FRs and / or constant region) are derived from a human immunoglobulin (acceptor antibody). Humanized antibodies generally retain the desired properties of the donor antibody, including, but not limited to, antigen specificity, affinity, reactivity, the ability to enhance immune cell activation, and the ability to enhance the immune response. The donor antibody may be a murine antibody, rat antibody, rabbit antibody, or non-human primate (e.g., cynomolgus monkey) antibody with the desired properties (e.g., antigen specificity, affinity, reactivity, the ability to enhance immune cell activation, and / or the ability to enhance the immune response).

[0121] Humanized antibodies are particularly useful because they can not only retain the desired properties of non-human donor antibodies (e.g., murine antibodies) but also effectively reduce the immunogenicity of non-human donor antibodies (e.g., murine antibodies) in human subjects. However, due to the lack of match between the CDRs of the donor antibody and the FRs of the recipient antibody, the desired properties of humanized antibodies (e.g., antigen specificity, affinity, reactivity, ability to improve immune cell activity and / or ability to enhance immune responses) are generally inferior to those of non-human donor antibodies (e.g., murine antibodies).

[0122] Thus, although researchers in the field have conducted in-depth research into antibody humanization and have achieved some success (see, for example, Jones et al., Nature, 321:522-525 (1986); Reichmann et al., Nature, 332:323-329 (1988); Presta, Curr. Op. Struct. Biol., 2:593-596 (1992); Clark, Immunol. Today 21:397-402 (2000)), the prior art did not provide detailed teachings on how to sufficiently humanize a particular donor antibody so that the resulting humanized antibody has as high a degree of humanization as possible while retaining as much of the desired properties of the donor antibody as possible. Without extensive inventive work by those skilled in the art, searching, testing, and improving upon a particular donor antibody, it is not possible to obtain a humanized antibody that is highly humanized (e.g., at least 75% humanized, 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% humanized) and retains the desired properties of the particular donor antibody.

[0123] In the present invention, in order to preserve the properties of the donor antibody (e.g., antigen specificity, affinity, reactivity, ability to improve immune cell activity and / or ability to enhance immune responses) as much as possible in the humanized antibody, the framework region (FR) of the humanized antibody of the present invention may contain amino acid residues of a human acceptor antibody or may contain amino acid residues of the corresponding non-human donor antibody.

[0124] As used herein, the term "degree of humanization" refers to an index for evaluating the number of non-human amino acid residues in a humanized antibody. The degree of humanization of a humanized antibody can be predicted by, for example, predicting the homology of the variable region sequence to the human V domain using DomainGapAlign on the IMGT website.

[0125] As used herein, the term "specifically bind" refers to a non-random binding reaction between two molecules, such as the reaction between an antibody and its target antigen. The strength or affinity of a specific binding interaction is determined by the equilibrium dissociation constant (K D ) or half-maximal inhibitory concentration (EC 50 ) may also be expressed as

[0126] The specific binding properties of two molecules may be measured by methods well known in the art. One such method involves measuring the formation and dissociation rates of an antigen-binding site or antigen complex. Both the "association rate constant" (ka or k) and the "dissociation rate constant" (k or k) can be calculated from the concentrations and the actual association and dissociation rates (see Malmqvist M, Nature, 1993, 361:186-187). The ratio of k to k is the dissociation constant, K D (See Davies et al., Annual Rev Biochem, 1990;59:439-473.) D The values of k, k, and k may be measured by any available method. In some embodiments, dissociation constants may be measured by biolayer interferometry (e.g., ForteBio Octet). Alternatively, dissociation constants may be measured by surface plasmon resonance (e.g., Biacore) or Kinexa.

[0127] As used herein, the term "vector" refers to a nucleic acid delivery vehicle into which a polynucleotide can be inserted. A vector is called an expression vector when it is capable of expressing a protein encoded by the inserted polynucleotide. A vector can be introduced into a host cell by transformation, transduction, or transfection to express the genetic material elements carried by the vector in the host cell. Vectors are well known to those skilled in the art and include, but are not limited to, plasmids, phagemids, cosmids, artificial chromosomes (e.g., yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), or P1-derived artificial chromosomes (PACs)), phages (e.g., lambda phage or M13 phage), and animal viruses. Animal viruses that can be used as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpes viruses (e.g., herpes simplex viruses), poxviruses, baculoviruses, papilloma viruses, and papova viruses (e.g., SV40). Vectors may contain a variety of expression control elements, including, but not limited to, promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, reporter genes, etc. Vectors may also contain an origin of replication site.

[0128] As used herein, the term "host cell" refers to a cell used to introduce a vector, including, but not limited to, prokaryotic cells such as E. coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus oryzae, insect cells such as Drosophila S2 cells or Sf9 cells, or animal cells such as fibroblasts, CHO cells, COS cells, NSO cells, HeLa cells, BHK cells, HEK 293 cells, or human cells. As used herein, the term "identity" refers to sequence identity between two polypeptides or two nucleic acids. To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are compared for optimal comparison (e.g., gaps may be introduced into the first amino acid or nucleic acid sequence to optimize comparison with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity of the two sequences is a function of the number of identical positions shared by the sequences (percent identity = number of identical overlapping positions / total number of positions x 100%). In some embodiments, the two sequences are the same length.

[0129] Determining the percent identity of two sequences can also be accomplished using a mathematical algorithm. A non-limiting example of a mathematical algorithm for comparing two sequences is the algorithm described in Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. USA 87:2264-2268, and its improved version in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90:5873-5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403.

[0130] As used herein, the term "variant," in the context of referring to polypeptides (including polypeptides), also refers to a polypeptide or peptide containing an altered amino acid sequence by introducing substitutions, deletions, or additions of amino acid residues. In some cases, the term "variant" also refers to a polypeptide or peptide that has been modified (e.g., by covalently attaching any type of molecule to the polypeptide or peptide). For example, but not limited to, a polypeptide may be modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protective blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Derivative polypeptides or peptides may also be produced by chemical modification using techniques known to those skilled in the art, including, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Variants also possess similar, identical, or improved functions as the polypeptide or peptide from which they are derived.

[0131] As used herein, the term "conservative substitution" refers to an amino acid substitution that does not adversely affect or alter the desired properties of a protein or polypeptide containing the amino acid sequence. For example, conservative substitutions may be introduced using standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include substitutions in which an amino acid residue is replaced with an amino acid residue having a similar side chain, e.g., substitution with a residue that is physically or functionally similar to the corresponding amino acid residue (e.g., has similar chemical properties such as size, shape, charge, or ability to form covalent or hydrogen bonds). Families of amino acid residues with similar side chains have been defined in the art. The family includes amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Therefore, it is preferable to replace a corresponding amino acid residue with another amino acid residue from the same side chain family. Amino acid residues may also be grouped into categories defined by any physical and functional properties. These include alcohol-containing residues (S, T), aliphatic residues (I, L, V, M), cycloalkenyl-related residues (F, H, W, Y), hydrophobic residues (A, C, F, G, H, I, L, M, R, T, V, W, Y), negatively charged residues (D, E), polar residues (C, D, E, H, K, N, Q, R, S, T), positively charged residues (H, K, R), small residues (A, C, D, G, N, P, S, T, V), small polar residues (A, G, S), residues involved in turn formation (A, C, D, E, G, H, K, N, Q, R, S, P, T), and flexible residues (Q, T, K, S, G, P, D, E, R).Methods for identifying conservative amino acid substitutions are well known in the art (see, e.g., Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10):879-884 (1999); Burks et al., Proc. Natl. Acad. Set USA 94:412-417 (1997), which are incorporated herein by reference).

[0132] The notation of the 20 common amino acids herein follows conventional convention. See, for example, Immunology-A Synthesis (2nd Edition, E.S. Golub and D.R. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In the present invention, the terms "polypeptide" and "protein" have the same meaning and are used interchangeably. Also, in the present invention, amino acids are generally represented by one-letter or three-letter abbreviations well known in the art. For example, alanine is represented by A or Ala.

[0133] As used herein, the term "linker" refers to a molecular moiety that links a conjugate moiety to an antibody or antigen-binding fragment thereof targeting a specific antigen to form a conjugate. In some cases, the linker further comprises a spacer to alter the steric hindrance of the conjugate moiety. While a linker is essential in some conjugates, a linker is optional in other conjugates. Linkers are classified into non-cleavable linkers and cleavable linkers. Conjugates containing non-cleavable linkers must be internalized to release the conjugate moiety by degrading the antibody or antigen-binding fragment thereof. Conjugates containing cleavable linkers may be internalized to release the conjugate moiety by degrading the antibody or antigen-binding fragment thereof, or may be non-internalized to release the conjugate moiety by disrupting the linker structure through enzymatic or chemical catalyzed cleavage at the cleavage site, thereby disrupting the linker structure. An example of a non-cleavable linker is N-succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC). Examples of the cleavable linker include a linker having a disulfide bond as the cleavage site, a hydrazone linker having an acyl group as the cleavage site, and a linker having an enzymatic catalytic site as the cleavage site (e.g., dipeptide linkers Val-Cit and Val-Ala, tetrapeptide linker Gly-Gly-Phe-Gly, and linkers containing phosphate and pyrophosphate).

[0134] The linker may link the antibody or antigen-binding fragment thereof to the conjugate moiety via a covalent bond through the reaction of chemical functional groups using conjugation techniques known in the art. The conjugation technique may be chemically specific in situ antibody modification, such as linkage achieved by the reaction of lysine residues with electrophilic groups, linkage achieved by the reaction of disulfide bonds, linkage achieved by re-crosslinking using sulfhydryl groups, or linkage achieved by glycosylation of glycans. The conjugation technique may also be site-specific bioconjugation based on genetically engineered antibodies, such as enzymatically catalyzed linkage achieved by adding natural or unnatural amino acids that are specifically recognized by enzymes to the antibody.

[0135] As used herein, the term "pharmaceutically acceptable carrier and / or excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with the subject and active ingredient, and is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995). Examples of suitable carriers and / or excipients include, but are not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, diluents, osmolality maintenance agents, absorption retarding agents, and preservatives. For example, pH adjusters include, but are not limited to, phosphate-buffered saline. 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 various antibacterial and antifungal agents, such as, but not limited to, methyl 4-hydroxybenzoate, chlorobutanol, phenol, sorbic acid, etc. Tonicity maintaining agents include, but are not limited to, sugars, NaCl, and analogs thereof. Absorption delaying agents include, but are not limited to, monostearic acid salts and gelatin. Diluents include, but are not limited to, water, aqueous buffer solutions (such as buffered saline), alcohols, and polyols (such as glycerol). Preservatives include various antibacterial and antifungal agents, such as, but not limited to, thimerosal, 2-phenoxyethanol, methyl 4-hydroxybenzoate, chlorobutanol, phenol, sorbic acid, etc.The stabilizer has the general meaning understood by those skilled in the art, and is capable of stabilizing the desired activity of the active ingredient in the drug, and includes, but is not limited to, sodium glutamate, gelatin, SPGA, sugars (sorbitol, mannitol, starch, sucrose, lactose, glucan, glucose, etc.), amino acids (glutamic acid, glycine, etc.), proteins (dried whey, albumin, casein, etc.) or degradation products thereof (lactalbumin hydrolysate, etc.), etc.

[0136] As used herein, the term "prophylaxis" refers to a method performed to prevent or delay the onset of a disease, condition, or symptom in a subject. As used herein, the term "treatment" refers to a method performed to obtain beneficial or desired clinical results. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, whether detectable or undetectable, reduction in the extent of disease, stabilization of the disease state (without further worsening), delay or mitigation of disease progression, improvement or alleviation of the disease state, reduction or inhibition of disease recurrence, and alleviation of symptoms (whether partial or total). "Treatment" can also refer to prolonging survival compared to desired survival (in the absence of treatment).

[0137] As used herein, the term "subject" refers to a mammal, e.g., a primate mammal, e.g., a human. In some embodiments, the subject (e.g., a human) has or is at risk for a tumor (e.g., a tumor that expresses MSLN), an inflammatory disease, or an autoimmune disease.

[0138] As used herein, the term "effective amount" refers to an amount sufficient to achieve or at least partially achieve a desired effect. For example, an effective amount for preventing a disease (e.g., a tumor, an inflammatory disease, or an autoimmune disease) refers to an amount sufficient to prevent, inhibit, or delay the onset of the disease (e.g., a tumor, an inflammatory disease, or an autoimmune disease), and an effective amount for treating a disease refers to an amount sufficient to cure or at least partially prevent the disease and its complications in a patient suffering from the disease. Determining such effective amounts is entirely within the capabilities of those skilled in the art. For example, an effective amount for therapeutic use will depend on the severity of the disease being treated, the overall state of the patient's own immune system, the patient's general condition (e.g., age, weight, sex), the mode of drug administration, other treatments administered simultaneously, and the like.

[0139] The terms "cancer" and "tumor" are used interchangeably to refer to a category of diseases characterized by the uncontrolled growth of abnormal cells in the body. Uncontrolled cell division can lead to the formation of malignant tumors or cells that invade neighboring tissues, which may metastasize to distant parts of the body via the lymphatic system or bloodstream. Cancer includes benign and malignant cancers, dormant tumors, or micrometastases. Cancer also includes hematologic malignancies.

[0140] As used herein, the term "pharmaceutically acceptable" refers to a molecule, molecule fragment, or composition that does not produce any adverse, allergic, or other toxic effects when administered appropriately to an animal or human. Specific examples of some substances that can be pharmaceutically acceptable vectors or components thereof include sugars (e.g., lactose), starch, cellulose and its derivatives, vegetable oils, gelatin, polyols (e.g., propylene glycol), alginic acid, etc.

[0141] As used herein, combination therapy includes the combination of an anti-CDCP1 antibody or antigen-binding fragment thereof of the present invention with one or more additional active therapeutic agents of a second therapy (e.g., a chemotherapeutic agent) or other prophylactic or therapeutic measures (e.g., radiation therapy).

[0142] Exemplary anti-cancer agents for second therapy may include chemotherapeutic agents (e.g., antimitotic agents), alkylating agents (e.g., Nitrogen Mustard), antimetabolites (e.g., folic acid analogs), natural products (e.g., Vinca Alkaloids), various reagents (e.g., platinum complexes), hormones and antagonists (e.g., corticosteroids), immunomodulators (e.g., Bropirimine (Upjohn)), etc. Other anti-cancer treatments include specific antibodies that target other cancer cells.

[0143] In such combination therapies, the various active agents always have different, complementary mechanisms of action, which can result in synergistic effects. Combination therapies include therapeutic agents that affect the immune response (e.g., enhance or activate the response) and therapeutic agents that affect tumor or cancer cells (e.g., suppress or kill them). Combination therapies can reduce the likelihood of developing drug-resistant cancer cells. Combination therapies can reduce the dose of one or more of the reagents, thereby reducing or eliminating adverse events related to one or more of the reagents. Such combination therapies can provide a synergistic therapeutic or preventative effect for the underlying disease, symptom, or condition.

[0144] As used herein, "combination" includes therapies that are administered separately, e.g., formulated separately for separate administration (e.g., provided as a kit), and therapies that are administered simultaneously as a single formulation (a "co-formulation"). In some embodiments, the anti-CDCP1 antibodies or antigen-binding fragments thereof of the present invention may be administered sequentially. In other embodiments, the anti-CDCP1 antibodies or antigen-binding fragments thereof may be administered simultaneously. The anti-CDCP1 antibodies or antigen-binding fragments thereof of the present invention may be used in any combination with at least one other (active) agent. [Effects of the Invention]

[0145] (Beneficial effects of the invention) The anti-CDCP1 antibodies provided by the present invention have clear advantages over known anti-CDCP1 antibodies, such as excellent target binding ability, tumor cell binding ability, and endocytosis ability, as well as better tumor cell migration inhibition ability and better drug development potential. The present invention also provides anti-CDCP1 antibody conjugates capable of targeting CDCP1-positive cells. The antibody conjugates of the present invention can target CDCP1-positive cells with high affinity and specificity, making them extremely useful for cancer treatment or in vivo diagnosis. The Examples demonstrate that the ADCs of the present invention have potent killing activity against various tumor cell lines expressing human CDCP1, such as colorectal cancer, breast cancer, and prostate cancer. Furthermore, in a mouse colorectal cancer xenograft tumor model, the ADCs of the present invention exhibit significant tumor-suppressing activity and have favorable safety profiles.

[0146] Hereinafter, the embodiments of the present invention will be described in detail with reference to the drawings and examples. However, it should be understood by those skilled in the art that the drawings and examples are for illustrative purposes only and do not limit the scope of the present invention. It is believed that the various objectives and advantageous aspects of the present invention will be realized by those skilled in the art from the following detailed description with reference to the drawings and preferred embodiments. [Brief explanation of the drawings]

[0147] [Figure 1] Figure 1 is a schematic diagram of the structures of CDCP1-C, CDCP1-A, and CDCP1-B. [Figure 2] FIG. 2 shows the results of detecting the binding ability of anti-CDCP1 antibodies to tumor cells. [Figure 3] Figure 3 shows the results of laser confocal detection of antibody internalization in tumor cell lines. In the figure, green represents the anti-CDCP1 antibody, red represents lysosomes, and arrows indicate their co-localization within the cells after endocytosis. [Figure 4a] FIG. 4a shows the effect of anti-CDCP1 antibodies on the proliferation of MDA-MB-231 tumor cells. [Figure 4b]FIG. 4b shows the effect of anti-CDCP1 antibodies on the proliferation of PC3 tumor cells. [Figure 4c] FIG. 4c shows the effect of anti-CDCP1 antibodies on the migration ability of tumor cells. [Figure 5] FIG. 5 shows the inhibitory effect of anti-CDCP1 chimeric antibody-vcMMAE conjugate on tumor cell growth. [Figure 6a] Figure 6a shows the inhibitory effect of anti-CDCP1 humanized antibody-vcMMAE conjugate on the growth of HCC1806 tumor cells. [Figure 6b] Figure 6b shows the inhibitory effect of anti-CDCP1 humanized antibody-vcMMAE conjugate on the growth of HCT116 tumor cells. [Figure 7a] FIG. 7a shows the effect of the anti-CDCP1 chimeric antibody-vcMMAE conjugate on mouse body weight. [Figure 7b] Figure 7b shows the inhibitory effect of the anti-CDCP1 chimeric antibody-vcMMAE conjugate on tumor growth in mice. [Example]

[0148] The invention will now be described by way of examples which are intended to illustrate, but not limit, the invention.

[0149] Sequence information relevant to the present invention is set forth in Table 1. [Table 1] JPEG2025525313000003.jpg238169 JPEG2025525313000004.jpg239169 JPEG2025525313000005.jpg239169 JPEG2025525313000006.jpg238169 JPEG2025525313000007.jpg239169 JPEG2025525313000008.jpg238169 JPEG2025525313000009.jpg238169 JPEG2025525313000010.jpg239169 JPEG2025525313000011.jpg238169 JPEG2025525313000012.jpg244169 JPEG2025525313000013.jpg239169 JPEG2025525313000014.jpg239169 JPEG2025525313000015.jpg244169 JPEG2025525313000016.jpg237169 JPEG2025525313000017.jpg230169 JPEG2025525313000018.jpg229169 JPEG2025525313000019.jpg236169 JPEG2025525313000020.jpg224169 JPEG2025525313000021.jpg223169 JPEG2025525313000022.jpg229169 JPEG2025525313000023.jpg217169 JPEG2025525313000024.jpg230169 JPEG2025525313000025.jpg230169 JPEG2025525313000026.jpg230169 JPEG2025525313000027.jpg230169 JPEG2025525313000028.jpg230169 JPEG2025525313000029.jpg229169 JPEG2025525313000030.jpg236169 JPEG2025525313000031.jpg237169 JPEG2025525313000032.jpg230169 JPEG2025525313000033.jpg230169 JPEG2025525313000034.jpg243169 JPEG2025525313000035.jpg238169 JPEG2025525313000036.jpg239169 JPEG2025525313000037.jpg46169

[0150] Unless otherwise specified, the molecular biological experimental methods and immunoassays used in the present invention are basically carried out with reference to the methods described in J. Sambrook et al., Molecular Cloning, A Laboratory Manual (Second Edition), Cold Spring Harbor Laboratory Press, 1989, and FMA Usubel et al., Short protocols in molecular biology, third edition, John Wiley & Sons, Inc., 1995, and restriction enzymes are used in accordance with the conditions recommended by the manufacturer. Those skilled in the art will recognize that the examples are provided to illustrate the present invention and are not intended to limit the scope of protection claimed by the present invention.

[0151] Example 1: Production of full-length human, cynomolgus monkey, rat, and mouse CDCP1-C extracellular domain antigens, and human CDCP1-A and CDCP1-B antigens In this example, CDCP1-C refers to the full-length extracellular domain (for example, in the case of human CDCP1, the amino acid sequence from the N-terminus to the C-terminus is the amino acid sequence from positions 30 to 667), CDCP1-A refers to the N-terminus, which is the membrane-proximal region of CDCP1 (for example, in the case of human CDCP1, the amino acid sequence from the N-terminus to the C-terminus is the amino acid sequence from positions 30 to 430), and CDCP1-B refers to the C-terminus, which is the membrane-proximal region of CDCP1 (for example, in the case of human CDCP1, the amino acid sequence from the N-terminus to the C-terminus is the amino acid sequence from positions 343 to 667). For specific structures, see Figure 1.

[0152] Human, cynomolgus monkey, rat, and mouse CDCP1 proteins were expressed in vitro. The gene numbers of the coding sequences of human CDCP1 protein, cynomolgus monkey CDCP1 protein, rat CDCP1 protein, and mouse CDCP1 protein were respectively NP_073753.3, XP_001114659.2, NP_001100339, and AAH85253. By molecular cloning, the corresponding full-length extracellular domain, membrane-extending region, or juxtamembrane region fragments were cloned into the expression vector pTT5, and then transfected into HEK-293T cells for expression. These were then purified using Fc / his tags. The purity and molecular weight were determined by SEC or SDS PAGE, and the purity was greater than 95% with an endotoxin content of less than 1 EU / mg. HuCDCP1-Fc, HuCDCP1-A, HuCDCP1-B, HuCDCP1-C, CynoCDCP1, RatCDCP1, and MsCDCP1 were obtained for subsequent animal immunization or screening experiments.

[0153] Example 2: Production of anti-CDCP1 murine monoclonal antibody BALBc mice were immunized with the human CDCP1-Fc (HuCDCP1-Fc) expressed in Example 1 once every two weeks. Mouse sera were collected after each immunization to detect the titer of anti-CDCP1 antibodies in the sera. After the fourth immunization, mouse splenocytes and SP2 / 0 cells were collected to perform hybridoma fusion. The hybridoma cells were plated on a 96-well plate. After 7 to 10 days, the supernatants were collected and coated with human CDCP1-his (Kactus Biosystems, CDC-HM101) for ELISA detection. Clone supernatants with an OD greater than 0.5 were selected and incubated with MDA-MB-231 cells (breast cancer cells highly expressing CDCP1, COBIOER BIOSCIENCES CO., LTD., CBP60382) at 4°C for 30 minutes. Furthermore, the clones were incubated with an anti-mFc secondary antibody (PE Goat anti-mouse IgG (minimal The cells were incubated with a secondary antibody (x-reactivity) antibody (Biolegend, 405307) and detected by flow cytometry. Cells in a group treated with only the secondary antibody, without adding hybridoma supernatant, served as a control. Using Geo-MFI (detection sample) / Geo-MFI (control) as a detection index, 298 hybridoma monoclonal antibodies with a 100-fold increase were selected. Next, antibody endocytosis experiments were performed on the screened hybridoma monoclonal antibodies, and hybridoma monoclonal antibodies with an endocytosis rate of greater than 50% in 1 hour were selected. The molecules were sorted through processes such as subcloning, affinity measurement, and sequencing. Finally, 33 hybridoma monoclonal antibodies were selected as candidate antibodies, designated 3B12H9, 3B12H5, and 3B12H6, respectively. , 3C11, 5G9, 4E12G10, 6A7, 6G9, 3F1A11, 20G4, 27D11, 3F1A11-H-12, 3A9, 19F7D6, 3E7, 5B8, 4B7, 8C9, 12E6, G10F6, 14A10C5, 8E5, 27A3, 10D2, 24G4, 1CP-061, 1CP-063, 1CP-062, 1CP-067, 1CP-069, 1CP-064, 1CP-066, 1CP-065, 1CP-068.

[0154] Example 3: Affinity measurement of anti-CDCP1 murine monoclonal antibody The HuCDCP1-A antigen, HuCDCP1-B antigen, HuCDCP1-C antigen, CynoCDCP1 antigen, MsCDCP1 antigen, and RatCDCP1 antigen (2 μg / mL) prepared in Example 1 were immobilized, respectively, and the binding status of the candidate antibody obtained in Example 2 and the control antibody CDSEQ135 (derived from sequence 135 in patent WO2011023389A1; SEQ135 and antibody 135 described in this application are both CDSEQ135 unless otherwise specified) was detected. Using Caterra or Biacore, the test antibody (the antibody was diluted to a maximum concentration of 150 nM, and then diluted 3-fold to five concentrations: 1.85 nM, 5.55 nM, 16.67 nM, 50 nM, and 150 nM) was flowed through the detection chip, respectively, and the data was analyzed using the corresponding software.

[0155] As shown in the detection results in Table 2, the 13 anti-CDCP1 murine monoclonal antibodies have good affinity for HuCDCP1-A, HuCDCP1-B, HuCDCP1-C, CynoCDCP1, MsCDCP1, or RatCDCP1. [Table 2]

[0156] Example 4: Measurement of binding ability of anti-CDCP1 murine monoclonal antibody to tumor cells The binding ability of the screened antibodies to tumor cells was measured using flow cytometry. MDA-MB-231 (breast cancer cells with high CDCP1 expression, COBIOER BIOSCIENCES CO., LTD., CBP60382), PC3 (prostate cancer cells with high CDCP1 expression, COBIOER BIOSCIENCES CO., LTD., CBP60343), MCF7 (breast cancer cells without CDCP1 expression, COBIOER BIOSCIENCES CO., LTD., CBP60380), and A2780 (human ovarian cancer cells without CDCP1 expression, COBIOER BIOSCIENCES CO., LTD., CBP60283) were digested with 5 mM EDTA and centrifuged. 2.0 × 10^4 tumor cells were added to each well. 50 μL of PBS + 2% FBS and an Fc blocker (Human TruStain) were added. After blocking with FcX™ (Biolegend, 422302) for 10 minutes at room temperature, anti-CDCP1 murine monoclonal antibody was added at a final concentration of 200 nM, diluted three-fold to eight concentrations, and incubated at 4°C for 30 minutes. The cells were then further incubated with a secondary antibody (PE anti-human IgG Fc Antibody, Biolegend, 410708) and analyzed by flow cytometry.

[0157] As shown in Table 3, the 11 anti-CDCP1 murine monoclonal antibodies showed good affinity to MDA-MB-231 and PC3 tumor cells, which highly express CDCP1, but did not bind to MCF7 and A2780 tumor cells, which do not express CDCP1. [Table 3]

[0158] Example 5: Measurement of endocytic activity of anti-CDCP1 murine monoclonal antibody The endocytic activity of antibodies was measured at 0.5 to 2 hours. MDA-MB-231 and PC3 cells were digested with 5 mM EDTA, counted, and plated at 4 x 10^4 cells / well. Fc blocker (Human TruStain FcX™, Biolegend, 422302) was added for 10 minutes at room temperature, followed by incubation with detection antibody (1.5 μg / mL) for 30 minutes at 4°C. The cells were then centrifuged and washed twice with PBS + 2% FBS. The cells were then divided into four equal portions (0, 0.5, 1, and 2 hours), and 200 μL of cell culture medium was added to each well and incubated at 37°C for 0.5, 1, and 2 hours, respectively. At time 0, cells were centrifuged and the supernatant removed. Secondary antibody (PE anti-human IgG Fc Antibody, Biolegend, 410708) was added and incubated at 4°C for 30 minutes. The cells were then centrifuged and washed twice with PBS + 2% FBS. Flow cytometry analysis was performed. After incubation at 37°C for 0.5, 1, and 2 hours, secondary antibody was added (using the same steps as at time 0). Geo-MFI, the signal measured at the same voltage, was then obtained. This signal was used as the signal detected at time 0. The antibody endocytosis rate was calculated using the formula: Internalization (%) = (Geo-MFI (0 time) - Geo-MFI (0.5, 1, and 2 hours)) / Geo-MFI (0 time) × 100%.

[0159] As shown in the detection results in Table 4, the murine monoclonal antibodies in the table exhibited good endocytosis activity for both MDA-MB-231 and PC3. [Table 4]

[0160] Example 6: Production of anti-CDCP1 antibodies by single B cell screening technology In this study, BALBc mice were immunized with the human CDCP1-Fc antigen expressed in Example 1 once every two weeks. Mouse serum was collected each time to detect the titer of anti-CDCP1-his antibody in the serum. After the fourth immunization, the mouse spleens were removed and processed using a commercially available kit (EasySep™ Mouse Pan-B Cell Isolation Kit, Stem Cell, 19844). The cells were counted and fluorescently labeled in vitro with B220 (B cell marker, PE anti-mouse / human CD45R / B220 Antibody, BioLegend, 103207) and CDCP1. B220+CDCP1+ cells were then selected by flow cytometry. Subsequently, the 10x Genomics kit (Chromium Next GEM Single Cell 5' Kit v2, 16 The antibodies were labeled with rxns (10xGENOMICS, PN-1000263), sequenced using single-cell BCRs, analyzed in vitro, classified by VH and VL sequences, and assigned to each family. Representative sequences were then selected and synthesized for each family. Finally, 18 candidate antibodies were selected, designated SB1, SB2, SB3, SB4, SB5, SB6, SB7, SB8, SB9, SB10, SB11, SB12, SB13, SB14, SB15, SB16, SB17, and SB18.

[0161] Example 7: Affinity measurement of anti-CDCP1 antibodies by single B cell screening technology The candidate antibodies obtained in Example 6 were cloned into an expression vector and expressed in small amounts in CHO cells. They were purified using an Fc affinity column (AT Protein A Diamond Plus, Catalog No. AA402312). The antibody purity was analyzed by HPLC, measured using Nanodrop, and high-throughput affinity assays were performed on small amounts of antibody using a Caterra. Specifically, the antibody binding status was detected by immobilizing the antibody with human CDCP1 antigen (2 μg / mL). Using a Caterra, the test antibody (diluted to a maximum concentration of 150 nM, then diluted 3-fold to five concentrations: 1.85 nM, 5.55 nM, 16.67 nM, 50 nM, and 150 nM) was run through the detection chip, and the data were analyzed using Kinetics software. Simultaneously, the antibody was coated with 2 μg / mL human CDCP1-C antigen and 2 μg / mL human CDCP1-B, and the antigen-binding ability of the antibody was measured by ELISA.

[0162] As shown in the detection results in Table 5, the anti-CDCP1 murine monoclonal antibodies in the table exhibit good affinity for HuCDCP1-C. [Table 5]

[0163] Example 8: Measurement of binding ability of anti-CDCP1 antibodies to tumor cells by single B cell screening technology The binding ability of the screened antibodies to tumor cells was measured using flow cytometry. MDA-MB-231 cells were digested with 5 mM EDTA and centrifuged. After cell counting, the cells were resuspended in FACS buffer (PBS + 2% FBS) and adjusted to a cell density of 4 x 10^5 / mL. 50 μL (2 x 10^4 tumor cells) was added to each well and blocked with Fc blocker (Human TruStain FcX™, Biolegend, 422302) at room temperature for 10 minutes. The test antibody was added to a final concentration of 1.5 μg / mL and incubated at 4°C for 30 minutes. The cells were then centrifuged, washed, and incubated with a secondary antibody (PE anti-human IgG Fc Antibody, Biolegend, 410708). The Geo-MFI was then measured by flow cytometry. The primary criterion for antibody cell binding was the antibody binding activity.

[0164] As shown in the detection results in FIG. 2, SB1 to SB15 and SB17 have good binding ability to MDA-MB-231 cells.

[0165] Example 9: Measurement of endocytic ability of anti-CDCP1 antibodies by single B cell screening technology The endocytic activity of antibodies was measured after 0.5 to 2 hours. MDA-MB-231 cells were digested with 5 mM EDTA, counted, and plated at 2 x 10^4 cells / well. They were blocked with Fc blocker (Human TruStain FcX™, Biolegend, 422302) for 10 minutes at room temperature. Detection antibodies (1.5 μg / mL) were added and incubated at 4°C for 30 minutes. The cells were centrifuged and washed twice with PBS + 2% FBS. The cells were then divided into four equal portions (0, 0.5, 1, and 2 hours), added to 200 μL of cell culture medium, and incubated at 37°C for 0.5, 1, and 2 hours, respectively. At time 0, the cells were centrifuged, and a secondary antibody (PE anti-human IgG Fc Antibody, Biolegend, 410708) was added. The cells were incubated at 4°C for 30 minutes, centrifuged, washed twice with PBS + 2% FBS, and analyzed by flow cytometry. After incubation at 37°C for 0.5, 1, and 2 hours, the secondary antibody (same steps as at time 0) was added and the signal at the same voltage, Geo-MFI, was obtained. This was used as the detection signal at time 0. The antibody endocytosis rate was calculated using the formula: internalization (%) = (Geo-MFI (0 time) - Geo-MFI (2 hours)) / Geo-MFI (0 time) × 100%.

[0166] As shown by the detection results in Table 6, MDA-MB-231 exhibits good endocytosis activity for all of the antibodies in Table 4. [Table 6]

[0167] Example 10: Identification of subtypes of anti-CDCP1 murine antibodies and amplification of variable regions The anti-CDCP1 murine monoclonal antibodies obtained in Example 2 and the anti-CDCP1 antibodies obtained by the single B cell screening technique in Example 6 were subcloned and cultured to obtain 5,000–1,000 monoclonal cells, which were then centrifuged. RNA was extracted from a small sample using the TaKaRa MiniBEST Universal RNA Extraction Kit (TaKaRa, cat. #9767). The extracted RNA was reverse transcribed using PrimeScript™ IV 1st strand cDNA Synthesis Mix (TaKaRa, cat. #6215A) to obtain cDNA, which was then amplified by PCR using VH and VL primers and ligated into the pTT5 vector. Five to ten clones were then selected and sequenced. The sequencing results were analyzed to define the CDR regions and locate the IgG family genes, allowing each antibody family to be distinguished and classified. The variable region and CDR sequences of the anti-CDCP1 murine monoclonal antibodies shown in Table 7 were obtained. [Table 7] JPEG2025525313000044.jpg89169

[0168] Example 11: Humanization of anti-CDCP1 murine antibody The anti-CDCP1 murine monoclonal antibodies 3C11, 8E5, and 8C9 were humanized. Specifically, the murine antibody sequences were compared with the amino acid sequence of human germline antibodies to identify sequences with high homology and excellent physical and chemical properties, which were used as the framework sequences for the humanized antibodies. The murine antibody CDR regions were defined using the IMGT method, and then grafted onto human antibody framework sequences. Sequence analysis and 3D simulation were used to restore some of the amino acids in the CDR and framework regions, as well as amino acids that may interact on the antibody surface. Furthermore, we calculated mutation energy, best-single mutations, germline substitutions, and frequent residue substitutions using a computer. We then measured the primary affinity of 12 mutants generated from each antibody. High-affinity and highly humanized antibodies were further expressed and tested for cell binding and endocytosis against multiple antigen genera. Of these, the 12 humanized antibodies produced from 8C9 were 1CP-151 to 1CP-162, the 12 humanized antibodies produced from 8E5 were 1CP-112 to 1CP-123, and the 12 humanized antibodies produced from 3C11 were 1CP-124 to 1CP-135.

[0169] The amino acid sequences of the CDRs of murine monoclonal antibodies 3C11, 8E5, and 8C9 are shown in Table 8. [Table 8]

[0170] The amino acid sequences of the variable and constant regions of the humanized antibody are shown in Table 9. [Table 9]

[0171] Example 12: Affinity measurement of humanized anti-CDCP1 antibodies The humanized anti-CDCP1 antibody obtained in Example 11 was cloned into a pTT5 expression vector and expressed in small amounts in CHO cells (Expi-CHOS, Gibco A29127). It was purified using an Fc affinity column (AT Protein A Diamond Plus, Catalog No. AA402312). The antibody purity was analyzed by HPLC, measured using a Nanodrop™, and high-throughput affinity assays were performed on small amounts of the antibody using a Caterra™. Specifically, the antibody binding status was detected by immobilizing the antibody with HuCDCP1-C antigen (2 μg / mL). Using a Caterra™, the test antibody (diluted to a maximum concentration of 150 nM, then diluted 3-fold to five concentrations: 1.85 nM, 5.55 nM, 16.67 nM, 50 nM, and 150 nM) was run through a detection chip, and the data were analyzed using Kinetics software.

[0172] As shown by the detection results in Table 10, the anti-CDCP1 humanized antibodies in the table exhibit good affinity for HuCDCP1-C. [Table 10]

[0173] Furthermore, some anti-CDCP1 humanized antibodies were selected and expressed, and then their affinities for the antigens Hu-CDCP1-A, Hu-CDCP1-B, Cyno-CDCP1, and Ms-CDCP1 were measured using the affinity measurement method described above. The antibodies in Table 11 were found to have good affinity for human and monkey CDCP1, and some antibodies exhibited the ability to bind to mouse CDCP1. [Table 11]

[0174] Example 13: Measurement of binding ability of anti-CDCP1 humanized antibodies to tumor cells According to the method of Example 4, MDA-MB-231 cells were used. The cells were digested with 5 mM EDTA, and 2 x 10^4 tumor cells were added to each well. 50 μL (PBS + 2% FBS) was added, and Fc blocker was added for 10 minutes at room temperature. Anti-CDCP1 humanized antibody was added to a final concentration of 200 nM, diluted 3-fold to eight concentrations, and incubated at 4°C for 30 minutes. The cells were centrifuged, washed, and incubated with a secondary antibody (PE anti-human IgG Fc antibody, Biolegend, 410708). Flow cytometry was performed to measure the cell binding affinity of the antibody. The Geo-MFI (geometric mean fluorescence signal) was used as the primary signal for antibody cell binding. GraphPad was used to calculate the cell binding affinity. As can be seen from the results in Table 12, the humanized antibodies in Table 12 have good cell binding affinity to MDA-MB-231. [Table 12]

[0175] Example 14: Measurement of endocytosis ability of anti-CDCP1 humanized antibodies The endocytosis ability of anti-CDCP1 humanized antibodies was measured according to Example 5. MDA-MB-231 cells were digested with 5 mM EDTA, counted, and plated at 3 x 10^4 cells / well. Fc blocker was added for 10 minutes at room temperature to block the cells. Detection antibody (1.5 μg / mL) was added and incubated at 4°C for 30 minutes. The cells were centrifuged and washed twice with PBS + 2% FBS. The cells were divided into three equal portions (0 h, 1 h, and 2 h). The 1- and 2-h portions were added to 200 μL of cell culture medium and incubated at 37°C for 1 and 2 hours, respectively. The 0-h portion was centrifuged, and a secondary antibody (PE anti-human IgG Fc antibody, Biolegend, 410708) was added. The cells were incubated at 4°C for 30 minutes. The secondary antibody was then centrifuged and washed twice with PBS + 2% FBS. The endocytosis ability was measured by flow cytometry. After incubation of the cells at 37°C for 1 and 2 hours, the secondary antibody (same steps as at time 0) was added and the signal, Geo-MFI, was obtained under the same voltage. This was used as the detection signal at time 0. The antibody endocytosis rate was calculated using the formula: internalization (%) = (Geo-MFI (0 hour) - Geo-MFI (1 hour, 2 hours)) / Geo-MFI (0 hour) × 100%.

[0176] As shown by the detection results in Table 13, the humanized antibody has good endocytosis activity at the cellular level. [Table 13]

[0177] Example 15: Laser confocal detection of antibody internalization in tumor cell lines 8E5, 10D2, 3B12H5, 3C11, 8C9, G10F6, 5B8, 1CP-063, 1CP-064, 1CP-062, 1CP-065, SB5, SB11, and SB14 were directly labeled with green fluorescent dye and then incubated with HCC1806 tumor cells (breast cancer cells with high CDCP1 expression). The antibody binding to the cell membrane and intracellular trafficking were monitored at 0, 5, 15, 30, 60, and 90 minutes using a laser confocal microscope. Lysosomes were labeled with red fluorescent dye, and the relative positioning and colocalization of the green and red fluorescent molecules within the cells was used to determine whether the antibodies colocalized with lysosomes after endocytosis.

[0178] The results of the antibody detection in this experiment are consistent with those shown in Figure 3, demonstrating that the detection antibody can clearly bind to the cell surface, which is consistent with the antibody cell binding measured by flow cytometry. Over time, the green fluorescent signal within the cells gradually strengthened, indicating that the green fluorescent signal on the cell surface gradually translocated into the cells and the antibody was internalized. The orange fluorescent signal gradually strengthened (the overlap of the green and red signals, indicated by the arrow), indicating that the antibody had been internalized and entered the lysosome. Therefore, the antibody provided by the present invention can enter the lysosome during endocytosis of tumor cells.

[0179] Example 16: Measurement of the effect of anti-CDCP1 antibody on proliferation and migration of tumor cell lines 3000 cells (breast cancer MDA-MB-231 cells and prostate cancer PC-3 cells) were plated in a 96-well plate overnight. Antibody concentrations ranging from 500 to 0.00128 nM (100 to 0.000256 nM, respectively) were added and diluted 5-fold to nine detection concentrations. The cells were incubated in a 37°C CO2 incubator for 96 hours. Half of the medium (100 μL) was discarded, and 100 μL of CellTiter-Glo Luminescent Cell Viability Assay Reagent (Promega, cat# G7570) was added to a BMG microplate reader for signal detection. The antibody inhibition of tumor cell growth was calculated by fitting and plotting the signal values and antibody concentrations using GraphPad. As can be seen from Figures 4a and 4b, the test antibodies 8E5, G10F6, 5B8, 8C9, 3C11, 3B12H5, 10D2, SEQ135, 14A10C5-2, 3F1A115-1, 3F1A115-2, 19F7D6, and 6A7 did not affect tumor cell growth.

[0180] In this study, to investigate the effect of antibodies on tumor cell metastasis, we performed a transwell experiment to detect tumor cell migration. Specifically, MDA-MB-231 cells were cultured and harvested, washed once with PBS, and adjusted to a density of 8 x 10^5 cells / mL. The test antibodies were added to the cells at a working concentration of 10 μg / mL and incubated for 30 minutes. The cell suspension was then added to the upper chamber of the transwell chamber and placed in an incubator for 12-48 hours. The chamber (transwell chamber for examining cell migration) was then removed, and the cells were fixed and stained with crystal violet for 15 minutes. The chamber was then washed, air-dried, sealed, and photographed under a microscope in 4-5 fields. The number of stained cells or the cell area were counted using ImageJ analysis software, which served as the basis for determining tumor cell migration.

[0181] As shown in the results in Figure 4c, the tested antibodies 38E11, 14A10C5, 5B8, 10D2, 8C9, G10F6, 8E5, 3C11, 3B12H5 and 6A7 have better tumor cell migration inhibitory abilities than SEQ135 and are therefore less likely to cause tumor migration-related adverse events.

[0182] The results showed that the tested antibodies did not affect tumor cell proliferation, while the control antibody SEQ135 caused tumor cell migration. However, the antibodies discovered in this study not only did not exhibit the ability to promote tumor cell migration, but also had some ability to inhibit tumor metastasis.

[0183] Example 17: Conjugation of anti-CDCP1 antibody with vcMMAE The anti-CDCP1 chimeric antibody is conjugated to MMAE via a vc linker. The vc-MMAE structure is as follows: [ka] Here, the structure of the vc linker is as follows: [ka]

[0184] Specifically, each antibody and the control antibody CD27H10 (antibody 27H10 described in WO2018112334A1) were reduced with 2x molar tris(2-chloroethyl)phosphate (TECP) for 1 hour at 4°C. Next, 7x molar vcMMAE was added and conjugated at 4-25°C for 1 hour. The reaction mixture was then dialyzed against 1x PBS (pH 8.0), filtered, and sterilized. The drug-antibody ratio (DAR) was measured as the hydrophobic peak by HPLC, and the molecular conjugation results are shown in Table 14. [Table 14]

[0185] Example 18: In vitro cancer cell killing assay by chimeric antibody-vcMMAE conjugates The cell-killing activity of the chimeric antibody ADC molecule was measured in CDCP1-expressing colorectal cancer cells HCT116 (COBIOER BIOSCIENCES CO., LTD, CBP60028), breast cancer cells MDA-MB-231 (COBIOER BIOSCIENCES CO., LTD, CBP60382), prostate cancer cells PC3 (COBIOER BIOSCIENCES CO., LTD, CBP60343), and CDCP1-nonexpressing MCF7 cells (COBIOER BIOSCIENCES CO., LTD, CBP60380). Specifically, cells were counted for each of the colorectal cancer cells HCT116, breast cancer cells MDA-MB-231, prostate cancer cells PC3, and MCF7 cells, adjusted to 3,000 cells / well, and plated overnight in a 96-well plate. The final concentrations of the conjugate added ranged from 8 to 0.000512 nM (diluted 5-fold to achieve seven detection concentrations), and the total volume was 200 μL. The cells were incubated in a 37°C CO2 incubator for 120 hours. Half of the medium (100 μL) was discarded, and 100 μL of CellTiter-Glo Luminescent Cell Viability Assay Reagent (Promega, cat# G7570) was added to a BMG microplate reader for signal detection. The inhibitory activity of the antibody against tumor cell growth was calculated by fitting and plotting the signal values and antibody concentrations using GraphPad. The results are shown in Figure 5 and Table 15. [Table 15]

[0186] The results showed that the tested chimeric antibody ADC molecules could effectively kill PC3, HCT116, and MDA-MB-231 tumor cells that express CDCP1, but had almost no killing effect on MCF7 cells that do not express CDCP1.

[0187] Example 19: In vivo inhibition of human colorectal cancer HCT116 tumor growth by chimeric antibody-vcMMAE conjugates in a mouse model HCT116 cells were cultured in McCoy's 5a medium + 10% FBS + 1% PS (Penicillin Streptomycin). HCT116 cells in the logarithmic growth phase were harvested and resuspended in McCoy's 5a medium to give 5 × 10 7 The tumor was subcutaneously inoculated into BALB / c nude mice (purchased from GemPharmatech LLC.) at a concentration of 100 cells / mL.

[0188] To create the animal model, 150 BALB / c nude mice were housed and adapted to the laboratory environment for 3 to 7 days. HCT116 cells were resuspended in McCoy's 5a medium and cultured at 5 × 10 6 The cells were then subcutaneously inoculated into the right back of experimental mice at 0.1 mL per mouse, and tumor growth was monitored periodically. The tumors in the mice reached an average volume of 150-200 mm. 3 When the tumors reached a size of approximately 100 mm, they were randomly assigned to groups of 8 mice per group based on tumor size, ensuring that tumor volumes and weights were similar across groups. The day of group assignment was defined as day 1.

[0189] The mice were administered and observed. On the day of group allocation, CD27H10-MMAE, IgG1-MMAE, 5B8-MMAE, 10D2-MMAE, 8C9-MMAE, G10F6-MMAE, 8E5-MMAE, 3C11-MMAE, 3B12H5-MMAE, 6A7-MMAE, and 14A10C5-MMAE were diluted with saline and administered via the tail vein at a dose of 1.5 mg / kg, adjusted based on body weight (5 μL / g). The mice were administered twice weekly for a total of four doses. A saline vehicle group was also included. After tumor inoculation, tumor growth and the effects of the drug on the animals' daily behavior, including activity, food and water intake, weight gain or loss, eyes, hair coat, feces, and other abnormalities, were routinely monitored. Any clinical symptoms observed during the experiment were recorded as raw data. After the start of administration, mice's weights and tumor volumes were measured twice weekly. Tumor volume (mm 3 ) is calculated as V=1 / 2×(a×b 2 ) where a is the major axis and b is the minor axis.

[0190] As shown in Figure 7a, there was no significant change in the body weight of the mice after administration, and Figure 7b shows that the antibody-vcMMAE conjugate of the present invention has a better in vivo tumor growth inhibitory effect than the CD27H10-MMAE control.

[0191] Example 20: Comparison of druggability after conjugation of anti-CDCP1 antibodies to vc-MMAE The ICP152 antibody and the control antibody CD27H10 were each linked to vc-MMAE using the method of Example 18, with a target DAR of 4. The physical and chemical properties of the molecules were then detected and analyzed. The analytical methods were as follows: Thermal stability measurements: The denaturation temperature (Tm), aggregation temperature (Tagg), and hydrodynamic diameter of the ADC molecules were measured using the UNcle Multipurpose Protein Stability Analyzer. Tm was measured by intrinsic fluorescence spectroscopy and fitted with the BCM algorithm, Tagg was measured by static light scattering (SLS), and hydrodynamic diameter was measured by dynamic light scattering (DLS).

[0192] SEC purity determination: The purity of the ADC molecules was analyzed by high performance liquid chromatography (HPLC).

[0193] Liquid chromatograph: Shimadzu LC-2030C Plus Column: TSKgel G3000SWXL, 7.8 x 300 mm, 5 μm, TOSOH Mobile phase: 61mmol / L Na2HPO4, 39mmol / L NaH2PO4, 200mmol / L NaCl, pH6.8 Flow rate: 1.0mL / min Detection wavelength: UV 220nm Column temperature: 30℃ Sample chamber temperature: 10°C Loading amount: 20μg Isocratic elution run time: 30 minutes The results were quantitatively analyzed using the area percentage method. If automatic integration failed due to factors such as baseline drift, retention time change, or peak shape, the integration parameters were adjusted or the spectrum was manually integrated. However, the main peak (MP) was the monomer peak, the peaks with retention times earlier than the main peak were polymer peaks (HMW), and the peaks with retention times later than the main peak were low molecular weight impurity peaks (LMW).

[0194] Tm is the temperature at which 50% of the protein molecules unfold during thermal denaturation and indicates the conformational stability of the molecule. Comparing the results, ICP152-MMAE and CD27H10-MMAE have similar Tm values, indicating their similar conformational stability. Tagg is the temperature at which protein molecules begin to aggregate and indicates the colloidal stability of the protein. The data in Table 16 show that the Tagg value for 1CP152-MMAE is 64.1°C, while that for CD27H10-MMAE is 60.1°C, a significant difference of 4°C between the two. During heating, CD27H10-MMAE aggregated earlier than 1CP152-MMAE, indicating that 1CP152-MMAE has better colloidal stability. Furthermore, comparing the hydrodynamic diameters of the two molecules, 1CP152-MMAE has a smaller particle size. It was suggested that 1CP152-MMAE diffuses faster in solution, has stronger mutual repulsion, and has better long-term storage stability. [Table 16]

[0195] In addition, in this study, the 1CP152-MMAE and CD27H10-MMAE molecules were incubated at 2-8°C for different periods (0, 3, and 5 days) to examine the changes in purity of both molecules. As shown in Table 17, the purity of 1CP152-MMAE remained unchanged after incubation at 2-8°C for 5 days, while the purity of CD27H10-MMAE decreased by 0.5% after incubation at 2-8°C for 5 days. This indicates that 1CP152-MMAE has better long-term storage stability. [Table 17]

[0196] Example 21: In vitro cancer cell killing assay by anti-CDCP1 humanized antibody-vcMMAE conjugate Humanized antibodies ICP125, ICP117 and ICP152 were conjugated to obtain ADCs, and the conjugation procedure was the same as in Example 18. The molecular conjugation results were as shown in Table 18. [Table 18]

[0197] Cell counts were performed on colorectal cancer cells HCT116 and breast cancer cells HCC1806, respectively, and the cells were plated overnight in a 96-well plate at 3,000 cells / well. The final conjugate concentrations ranged from 200 to 0.000512 nM (diluted 5-fold to achieve nine detection concentrations). A total of 200 μL of conjugate was added. The cells were incubated in a 37°C CO2 incubator for 120 hours. Half of the medium (100 μL) was discarded, and 100 μL of CellTiter-Glo Luminescent Cell Viability Assay Reagent (Promega, cat# G7570) was added to a BMG microplate reader for signal detection. The inhibitory activity of the antibody against tumor cell growth was calculated by fitting and plotting the signal values and antibody concentrations using GraphPad. The results are shown in Figures 6a and 6b, and Tables 19 and 20. [Table 19] [Table 20]

[0198] The results demonstrated that the tested humanized ADC molecules had significant tumor cell-killing effects on both HCC1806 and HCT116 cells.

[0199] Although the specific embodiments of the present invention have been described in detail, it is understood by those skilled in the art that various modifications and changes to the details may be made based on all the teachings disclosed, and all of these modifications fall within the scope of protection of the present invention. The present invention is limited by the following claims and any equivalents thereof.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to CDCP1, The antibody or antigen-binding fragment thereof (1) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 11, CDR-H2 comprising the sequence shown in SEQ ID NO: 48, and CDR-H3 comprising the sequence shown in SEQ ID NO: 90; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 136, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 14, CDR-H2 comprising the sequence shown in SEQ ID NO: 52, and CDR-H3 comprising the sequence shown in SEQ ID NO: 95; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 140, CDR-L2 comprising the sequence shown in SEQ ID NO: 175, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; Or, (3) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 2, CDR-H2 comprising the sequence shown in SEQ ID NO: 39, and CDR-H3 comprising the sequence shown in SEQ ID NO: 81; and a VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 125, CDR-L2 comprising the sequence shown in SEQ ID NO: 166, and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; Or, (4) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 1, CDR-H2 comprising the sequence shown in SEQ ID NO: 38, and CDR-H3 comprising the sequence shown in SEQ ID NO: 80; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 124, CDR-L2 comprising the sequence shown in SEQ ID NO: 164, and CDR-L3 comprising the sequence shown in SEQ ID NO: 197; Or, (5) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 1, CDR-H2 comprising the sequence shown in SEQ ID NO: 38, and CDR-H3 comprising the sequence shown in SEQ ID NO: 80; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 125, CDR-L2 comprising the sequence shown in SEQ ID NO: 165, and CDR-L3 comprising the sequence shown in SEQ ID NO: 198; Or, (6) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 3; CDR-H2 comprising the sequence shown in SEQ ID NO: 39; and CDR-H3 comprising the sequence shown in SEQ ID NO: 81; A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 126, CDR-L2 comprising the sequence shown in SEQ ID NO: 167, and CDR-L3 comprising the sequence shown in SEQ ID NO: 200; Or, (7) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 4, CDR-H2 comprising the sequence shown in SEQ ID NO: 40, and CDR-H3 comprising the sequence shown in SEQ ID NO: 82; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 127, CDR-L2 comprising the sequence shown in SEQ ID NO: 168, and CDR-L3 comprising the sequence shown in SEQ ID NO: 201; Or, (8) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 5, CDR-H2 comprising the sequence shown in SEQ ID NO: 41, and CDR-H3 comprising the sequence shown in SEQ ID NO: 83; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 128, CDR-L2 comprising the sequence shown in SEQ ID NO: 169, and CDR-L3 comprising the sequence shown in SEQ ID NO: 202; Or, (9) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 6, CDR-H2 comprising the sequence shown in SEQ ID NO: 42, and CDR-H3 comprising the sequence shown in SEQ ID NO: 84; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 129, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (10) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 6, CDR-H2 comprising the sequence shown in SEQ ID NO: 43, and CDR-H3 comprising the sequence shown in SEQ ID NO: 85; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 130, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (11) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 7, CDR-H2 comprising the sequence shown in SEQ ID NO: 44, and CDR-H3 comprising the sequence shown in SEQ ID NO: 86; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 131, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 204; Or, (12) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 7, CDR-H2 comprising the sequence shown in SEQ ID NO: 44, and CDR-H3 comprising the sequence shown in SEQ ID NO: 86; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 132, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 204; Or, (13) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 7, CDR-H2 comprising the sequence shown in SEQ ID NO: 44, and CDR-H3 comprising the sequence shown in SEQ ID NO: 86; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 130, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (14) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 8, CDR-H2 comprising the sequence shown in SEQ ID NO: 45, and CDR-H3 comprising the sequence shown in SEQ ID NO: 87; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 133, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 203; Or, (15) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 9, CDR-H2 comprising the sequence shown in SEQ ID NO: 46, and CDR-H3 comprising the sequence shown in SEQ ID NO: 88; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 134, CDR-L2 comprising the sequence shown in SEQ ID NO: 164, and CDR-L3 comprising the sequence shown in SEQ ID NO: 205; Or, (16) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 10, CDR-H2 comprising the sequence shown in SEQ ID NO: 47, and CDR-H3 comprising the sequence shown in SEQ ID NO: 89; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 135, CDR-L2 comprising the sequence shown in SEQ ID NO: 172, and CDR-L3 comprising the sequence shown in SEQ ID NO: 206; Or, (17) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 12, CDR-H2 comprising the sequence shown in SEQ ID NO: 48, and CDR-H3 comprising the sequence shown in SEQ ID NO: 91; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 136, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (18) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 11, CDR-H2 comprising the sequence shown in SEQ ID NO: 48, and CDR-H3 comprising the sequence shown in SEQ ID NO: 90; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 136, CDR-L2 comprising the sequence shown in SEQ ID NO: 171, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (19) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13; CDR-H2 comprising the sequence shown in SEQ ID NO: 49; and CDR-H3 comprising the sequence shown in SEQ ID NO: 92; A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 137, CDR-L2 comprising the sequence shown in SEQ ID NO: 173, and CDR-L3 comprising the sequence shown in SEQ ID NO: 208; Or, (20) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13, CDR-H2 comprising the sequence shown in SEQ ID NO: 50, and CDR-H3 comprising the sequence shown in SEQ ID NO: 93; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 138, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 209; Or, (21) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13; CDR-H2 comprising the sequence shown in SEQ ID NO: 51; and CDR-H3 comprising the sequence shown in SEQ ID NO: 94; A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 139, CDR-L2 comprising the sequence shown in SEQ ID NO: 170, and CDR-L3 comprising the sequence shown in SEQ ID NO: 210; Or, (22) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 15, CDR-H2 comprising the sequence shown in SEQ ID NO: 53, and CDR-H3 comprising the sequence shown in SEQ ID NO: 96; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 124, CDR-L2 comprising the sequence shown in SEQ ID NO: 164, and CDR-L3 comprising the sequence shown in SEQ ID NO: 212; Or, (23) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 16, CDR-H2 comprising the sequence shown in SEQ ID NO: 54, and CDR-H3 comprising the sequence shown in SEQ ID NO: 97; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 141, CDR-L2 comprising the sequence shown in SEQ ID NO: 172, and CDR-L3 comprising the sequence shown in SEQ ID NO: 213; Or, (24) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 17, CDR-H2 comprising the sequence shown in SEQ ID NO: 55, and CDR-H3 comprising the sequence shown in SEQ ID NO: 98; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 126, CDR-L2 comprising the sequence shown in SEQ ID NO: 174, and CDR-L3 comprising the sequence shown in SEQ ID NO: 214; Or, (25) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 18, CDR-H2 comprising the sequence shown in SEQ ID NO: 56, and CDR-H3 comprising the sequence shown in SEQ ID NO: 99; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 142, CDR-L2 comprising the sequence shown in SEQ ID NO: 176, and CDR-L3 comprising the sequence shown in SEQ ID NO: 215; Or, (26) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 19, CDR-H2 comprising the sequence shown in SEQ ID NO: 57, and CDR-H3 comprising the sequence shown in SEQ ID NO: 100; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 143, CDR-L2 comprising the sequence shown in SEQ ID NO: 177, and CDR-L3 comprising the sequence shown in SEQ ID NO: 216; Or, (27) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 20, CDR-H2 comprising the sequence shown in SEQ ID NO: 58, and CDR-H3 comprising the sequence shown in SEQ ID NO: 101; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 144, CDR-L2 comprising the sequence shown in SEQ ID NO: 176, and CDR-L3 comprising the sequence shown in SEQ ID NO: 215; Or, (28) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 21, CDR-H2 comprising the sequence shown in SEQ ID NO: 59, and CDR-H3 comprising the sequence shown in SEQ ID NO: 102; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 127, CDR-L2 comprising the sequence shown in SEQ ID NO: 168, and CDR-L3 comprising the sequence shown in SEQ ID NO: 217; Or, (29) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 21, CDR-H2 comprising the sequence shown in SEQ ID NO: 60, and CDR-H3 comprising the sequence shown in SEQ ID NO: 102; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 127, CDR-L2 comprising the sequence shown in SEQ ID NO: 168, and CDR-L3 comprising the sequence shown in SEQ ID NO: 217; Or, (30) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 13, CDR-H2 comprising the sequence shown in SEQ ID NO: 49, and CDR-H3 comprising the sequence shown in SEQ ID NO: 103; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 144, CDR-L2 comprising the sequence shown in SEQ ID NO: 169, and CDR-L3 comprising the sequence shown in SEQ ID NO: 218; Or, (31) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 22, CDR-H2 comprising the sequence shown in SEQ ID NO: 61, and CDR-H3 comprising the sequence shown in SEQ ID NO: 104; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 145, CDR-L2 comprising the sequence shown in SEQ ID NO: 178, and CDR-L3 comprising the sequence shown in SEQ ID NO: 219; Or, (32) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 17, CDR-H2 comprising the sequence shown in SEQ ID NO: 62, and CDR-H3 comprising the sequence shown in SEQ ID NO: 105; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 141, CDR-L2 comprising the sequence shown in SEQ ID NO: 172, and CDR-L3 comprising the sequence shown in SEQ ID NO: 220; Or, (33) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 17, CDR-H2 comprising the sequence shown in SEQ ID NO: 63, and CDR-H3 comprising the sequence shown in SEQ ID NO: 106; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 126, CDR-L2 comprising the sequence shown in SEQ ID NO: 179, and CDR-L3 comprising the sequence shown in SEQ ID NO: 221; Or, (34) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 23, CDR-H2 comprising the sequence shown in SEQ ID NO: 64, and CDR-H3 comprising the sequence shown in SEQ ID NO: 107; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 146, CDR-L2 comprising the sequence shown in SEQ ID NO: 180, and CDR-L3 comprising the sequence shown in SEQ ID NO: 222; Or, (35) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 24, CDR-H2 comprising the sequence shown in SEQ ID NO: 65, and CDR-H3 comprising the sequence shown in SEQ ID NO: 108; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 147, CDR-L2 comprising the sequence shown in SEQ ID NO: 181, and CDR-L3 comprising the sequence shown in SEQ ID NO: 223; Or, (36) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 25, CDR-H2 comprising the sequence shown in SEQ ID NO: 65, and CDR-H3 comprising the sequence shown in SEQ ID NO: 108; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 148, CDR-L2 comprising the sequence shown in SEQ ID NO: 181, and CDR-L3 comprising the sequence shown in SEQ ID NO: 224; Or, (37) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 26, CDR-H2 comprising the sequence shown in SEQ ID NO: 66, and CDR-H3 comprising the sequence shown in SEQ ID NO: 109; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 149, CDR-L2 comprising the sequence shown in SEQ ID NO: 182, and CDR-L3 comprising the sequence shown in SEQ ID NO: 225; Or, (38) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 27, CDR-H2 comprising the sequence shown in SEQ ID NO: 67, and CDR-H3 comprising the sequence shown in SEQ ID NO: 110; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 150, CDR-L2 comprising the sequence shown in SEQ ID NO: 183, and CDR-L3 comprising the sequence shown in SEQ ID NO: 226; Or, (39) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 28, CDR-H2 comprising the sequence shown in SEQ ID NO: 68, and CDR-H3 comprising the sequence shown in SEQ ID NO: 111; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 151, CDR-L2 comprising the sequence shown in SEQ ID NO: 184, and CDR-L3 comprising the sequence shown in SEQ ID NO: 227; Or, (40) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 29, CDR-H2 comprising the sequence shown in SEQ ID NO: 69, and CDR-H3 comprising the sequence shown in SEQ ID NO: 112; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 152, CDR-L2 comprising the sequence shown in SEQ ID NO: 185, and CDR-L3 comprising the sequence shown in SEQ ID NO: 222; Or, (41) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 30, CDR-H2 comprising the sequence shown in SEQ ID NO: 70, and CDR-H3 comprising the sequence shown in SEQ ID NO: 113; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 153, CDR-L2 comprising the sequence shown in SEQ ID NO: 186, and CDR-L3 comprising the sequence shown in SEQ ID NO: 228; Or, (42) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 31, CDR-H2 comprising the sequence shown in SEQ ID NO: 71, and CDR-H3 comprising the sequence shown in SEQ ID NO: 114; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 154, CDR-L2 comprising the sequence shown in SEQ ID NO: 187, and CDR-L3 comprising the sequence shown in SEQ ID NO: 229; Or, (43) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 32, CDR-H2 comprising the sequence shown in SEQ ID NO: 72, and CDR-H3 comprising the sequence shown in SEQ ID NO: 115; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 155, CDR-L2 comprising the sequence shown in SEQ ID NO: 188, and CDR-L3 comprising the sequence shown in SEQ ID NO: 230; Or, (44) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 33, CDR-H2 comprising the sequence shown in SEQ ID NO: 73, and CDR-H3 comprising the sequence shown in SEQ ID NO: 116; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 156, CDR-L2 comprising the sequence shown in SEQ ID NO: 189, and CDR-L3 comprising the sequence shown in SEQ ID NO: 231; Or, (45) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 34, CDR-H2 comprising the sequence shown in SEQ ID NO: 74, and CDR-H3 comprising the sequence shown in SEQ ID NO: 117; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 157, CDR-L2 comprising the sequence shown in SEQ ID NO: 190, and CDR-L3 comprising the sequence shown in SEQ ID NO: 232; Or, (46) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 35, CDR-H2 comprising the sequence shown in SEQ ID NO: 75, and CDR-H3 comprising the sequence shown in SEQ ID NO: 118; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 158, CDR-L2 comprising the sequence shown in SEQ ID NO: 191, and CDR-L3 comprising the sequence shown in SEQ ID NO: 233; Or, (47) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 26, CDR-H2 comprising the sequence shown in SEQ ID NO: 76, and CDR-H3 comprising the sequence shown in SEQ ID NO: 119; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 159, CDR-L2 comprising the sequence shown in SEQ ID NO: 192, and CDR-L3 comprising the sequence shown in SEQ ID NO: 234; Or, (48) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 24, CDR-H2 comprising the sequence shown in SEQ ID NO: 77, and CDR-H3 comprising the sequence shown in SEQ ID NO: 120; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 160, CDR-L2 comprising the sequence shown in SEQ ID NO: 193, and CDR-L3 comprising the sequence shown in SEQ ID NO: 235; Or, (49) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 36, CDR-H2 comprising the sequence shown in SEQ ID NO: 78, and CDR-H3 comprising the sequence shown in SEQ ID NO: 121; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 161, CDR-L2 comprising the sequence shown in SEQ ID NO: 194, and CDR-L3 comprising the sequence shown in SEQ ID NO: 236; Or, (50) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 32, CDR-H2 comprising the sequence shown in SEQ ID NO: 79, and CDR-H3 comprising the sequence shown in SEQ ID NO: 122; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 162, CDR-L2 comprising the sequence shown in SEQ ID NO: 195, and CDR-L3 comprising the sequence shown in SEQ ID NO: 237; Or, (51) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 37, CDR-H2 comprising the sequence shown in SEQ ID NO: 79, and CDR-H3 comprising the sequence shown in SEQ ID NO: 123; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 163, CDR-L2 comprising the sequence shown in SEQ ID NO: 196, and CDR-L3 comprising the sequence shown in SEQ ID NO: 238; wherein the CDRs are defined by the IMGT numbering system: Preferably, the antibody or antigen-binding fragment thereof comprises: (1) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 392, CDR-H2 comprising the sequence shown in SEQ ID NO: 393, and CDR-H3 comprising the sequence shown in SEQ ID NO: 394; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 395, CDR-L2 comprising the sequence shown in SEQ ID NO: 396, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 377, CDR-H2 comprising the sequence shown in SEQ ID NO: 378, and CDR-H3 comprising the sequence shown in SEQ ID NO: 379; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 380, CDR-L2 comprising the sequence shown in SEQ ID NO: 381, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; Or, (3) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 362, CDR-H2 comprising the sequence shown in SEQ ID NO: 363, and CDR-H3 comprising the sequence shown in SEQ ID NO: 364; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 365, CDR-L2 comprising the sequence shown in SEQ ID NO: 366, and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; wherein the CDRs are defined by the Kabat numbering system: Or, (1) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 397; CDR-H2 comprising the sequence shown in SEQ ID NO: 398; and CDR-H3 comprising the sequence shown in SEQ ID NO: 394; A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 395, CDR-L2 comprising the sequence shown in SEQ ID NO: 396, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 382, CDR-H2 comprising the sequence shown in SEQ ID NO: 383, and CDR-H3 comprising the sequence shown in SEQ ID NO: 379; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 380, CDR-L2 comprising the sequence shown in SEQ ID NO: 381, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; Or, (3) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 367, CDR-H2 comprising the sequence shown in SEQ ID NO: 368, and CDR-H3 comprising the sequence shown in SEQ ID NO: 364; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 365, CDR-L2 comprising the sequence shown in SEQ ID NO: 366, and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; The CDRs are defined by the AbM numbering system: Or, (1) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 399, CDR-H2 comprising the sequence shown in SEQ ID NO: 400, and CDR-H3 comprising the sequence shown in SEQ ID NO: 394; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 395, CDR-L2 comprising the sequence shown in SEQ ID NO: 396, and CDR-L3 comprising the sequence shown in SEQ ID NO: 207; Or, (2) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 384, CDR-H2 comprising the sequence shown in SEQ ID NO: 385, and CDR-H3 comprising the sequence shown in SEQ ID NO: 379; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 380, CDR-L2 comprising the sequence shown in SEQ ID NO: 381, and CDR-L3 comprising the sequence shown in SEQ ID NO: 211; Or, (3) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 369, CDR-H2 comprising the sequence shown in SEQ ID NO: 370, and CDR-H3 comprising the sequence shown in SEQ ID NO: 364; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 365, CDR-L2 comprising the sequence shown in SEQ ID NO: 366, and CDR-L3 comprising the sequence shown in SEQ ID NO: 199; The CDRs are defined by the Chothia numbering system: Or, (1) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 401, CDR-H2 comprising the sequence shown in SEQ ID NO: 402, and CDR-H3 comprising the sequence shown in SEQ ID NO: 403; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 404, CDR-L2 comprising the sequence shown in SEQ ID NO: 405, and CDR-L3 comprising the sequence shown in SEQ ID NO: 406; Or, (2) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 386, CDR-H2 comprising the sequence shown in SEQ ID NO: 387, and CDR-H3 comprising the sequence shown in SEQ ID NO: 388; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 389, CDR-L2 comprising the sequence shown in SEQ ID NO: 390, and CDR-L3 comprising the sequence shown in SEQ ID NO: 391; Or, (3) A VH comprising the following three CDRs: CDR-H1 comprising the sequence shown in SEQ ID NO: 371, CDR-H2 comprising the sequence shown in SEQ ID NO: 372, and CDR-H3 comprising the sequence shown in SEQ ID NO: 373; and A VL comprising the following three CDRs: CDR-L1 comprising the sequence shown in SEQ ID NO: 374, CDR-L2 comprising the sequence shown in SEQ ID NO: 375, and CDR-L3 comprising the sequence shown in SEQ ID NO: 376; The CDRs are defined by the Contact numbering system.

2. the antibody or antigen-binding fragment thereof (1) VH of the sequence shown in SEQ ID NO: 239 and VL of the sequence shown in SEQ ID NO: 289 (2) VH of the sequence shown in SEQ ID NO: 239 and VL of the sequence shown in SEQ ID NO: 290 (3) VH of the sequence shown in SEQ ID NO: 240 and VL of the sequence shown in SEQ ID NO: 291 (4) VH of the sequence shown in SEQ ID NO: 241 and VL of the sequence shown in SEQ ID NO: 292 (5) VH of the sequence shown in SEQ ID NO: 242 and VL of the sequence shown in SEQ ID NO: 293 (6) VH of the sequence shown in SEQ ID NO: 243 and VL of the sequence shown in SEQ ID NO: 294 (7) VH of the sequence shown in SEQ ID NO: 244 and VL of the sequence shown in SEQ ID NO: 295 (8) VH of the sequence shown in SEQ ID NO: 245 and VL of the sequence shown in SEQ ID NO: 296 (9) VH of the sequence shown in SEQ ID NO: 246 and VL of the sequence shown in SEQ ID NO: 297 (10) VH of the sequence shown in SEQ ID NO: 247 and VL of the sequence shown in SEQ ID NO: 298 (11) VH of the sequence shown in SEQ ID NO: 248 and VL of the sequence shown in SEQ ID NO: 296 (12) VH of the sequence shown in SEQ ID NO: 249 and VL of the sequence shown in SEQ ID NO: 299 (13) VH of the sequence shown in SEQ ID NO: 250 and VL of the sequence shown in SEQ ID NO: 300 (14) VH of the sequence shown in SEQ ID NO: 251 and VL of the sequence shown in SEQ ID NO: 301 (15) VH of the sequence shown in SEQ ID NO: 252 and VL of the sequence shown in SEQ ID NO: 302 (16) VH of the sequence shown in SEQ ID NO: 253 and VL of the sequence shown in SEQ ID NO: 303 (17) VH of the sequence shown in SEQ ID NO: 254 and VL of the sequence shown in SEQ ID NO: 304 (18) VH of the sequence shown in SEQ ID NO: 255 and VL of the sequence shown in SEQ ID NO: 305 (19) VH of the sequence shown in SEQ ID NO: 256 and VL of the sequence shown in SEQ ID NO: 306 (20) VH of the sequence shown in SEQ ID NO: 257 and VL of the sequence shown in SEQ ID NO: 307 (21) VH of the sequence shown in SEQ ID NO: 258 and VL of the sequence shown in SEQ ID NO: 308 (22) VH of the sequence shown in SEQ ID NO: 259 and VL of the sequence shown in SEQ ID NO: 309 (23) VH of the sequence shown in SEQ ID NO: 260 and VL of the sequence shown in SEQ ID NO: 310 (24) VH of the sequence shown in SEQ ID NO: 261 and VL of the sequence shown in SEQ ID NO: 311 (25) VH of the sequence shown in SEQ ID NO: 262 and VL of the sequence shown in SEQ ID NO: 312 (26) VH of the sequence shown in SEQ ID NO: 263 and VL of the sequence shown in SEQ ID NO: 313 (27) VH of the sequence shown in SEQ ID NO: 264 and VL of the sequence shown in SEQ ID NO: 314 (28) VH of the sequence shown in SEQ ID NO: 265 and VL of the sequence shown in SEQ ID NO: 315 (29) VH of the sequence shown in SEQ ID NO: 266 and VL of the sequence shown in SEQ ID NO: 316 (30) VH of the sequence shown in SEQ ID NO: 267 and VL of the sequence shown in SEQ ID NO: 317 (31) VH of the sequence shown in SEQ ID NO: 268 and VL of the sequence shown in SEQ ID NO: 318 (32) VH of the sequence shown in SEQ ID NO: 269 and VL of the sequence shown in SEQ ID NO: 319 (33) VH of the sequence shown in SEQ ID NO: 270 and VL of the sequence shown in SEQ ID NO: 320 (34) VH of the sequence shown in SEQ ID NO: 271 and VL of the sequence shown in SEQ ID NO: 321 (35) VH of the sequence shown in SEQ ID NO: 272 and VL of the sequence shown in SEQ ID NO: 322 (36) VH of the sequence shown in SEQ ID NO: 273 and VL of the sequence shown in SEQ ID NO: 323 (37) VH of the sequence shown in SEQ ID NO: 274 and VL of the sequence shown in SEQ ID NO: 324 (38) VH of the sequence shown in SEQ ID NO: 275 and VL of the sequence shown in SEQ ID NO: 325 (39) VH of the sequence shown in SEQ ID NO: 276 and VL of the sequence shown in SEQ ID NO: 326 (40) VH of the sequence shown in SEQ ID NO: 277 and VL of the sequence shown in SEQ ID NO: 327 (41) VH of the sequence shown in SEQ ID NO: 278 and VL of the sequence shown in SEQ ID NO: 328 (42) VH of the sequence shown in SEQ ID NO: 279 and VL of the sequence shown in SEQ ID NO: 329 (43) VH of the sequence shown in SEQ ID NO: 280 and VL of the sequence shown in SEQ ID NO: 330 (44) VH of the sequence shown in SEQ ID NO: 281 and VL of the sequence shown in SEQ ID NO: 331 (45) VH of the sequence shown in SEQ ID NO: 282 and VL of the sequence shown in SEQ ID NO: 332 (46) VH of the sequence shown in SEQ ID NO: 283 and VL of the sequence shown in SEQ ID NO: 333 (47) VH of the sequence shown in SEQ ID NO: 284 and VL of the sequence shown in SEQ ID NO: 334 (48) VH of the sequence shown in SEQ ID NO: 285 and VL of the sequence shown in SEQ ID NO: 335 (49) VH of the sequence shown in SEQ ID NO: 286 and VL of the sequence shown in SEQ ID NO: 336 (50) VH of the sequence shown in SEQ ID NO: 287 and VL of the sequence shown in SEQ ID NO: 337 (51) VH of the sequence shown in SEQ ID NO: 288 and VL of the sequence shown in SEQ ID NO: 338 (52) VH of the sequence shown in SEQ ID NO: 339 and VL of the sequence shown in SEQ ID NO: 351 (53) VH of the sequence shown in SEQ ID NO: 339 and VL of the sequence shown in SEQ ID NO: 352 (54) VH of the sequence shown in SEQ ID NO: 339 and VL of the sequence shown in SEQ ID NO: 353 (55) VH of the sequence shown in SEQ ID NO: 340 and VL of the sequence shown in SEQ ID NO: 351 (56) VH of the sequence shown in SEQ ID NO: 340 and VL of the sequence shown in SEQ ID NO: 352 (57) VH of the sequence shown in SEQ ID NO: 340 and VL of the sequence shown in SEQ ID NO: 353 (58) VH of the sequence shown in SEQ ID NO: 341 and VL of the sequence shown in SEQ ID NO: 351 (59) VH of the sequence shown in SEQ ID NO: 341 and VL of the sequence shown in SEQ ID NO: 352 (60) VH of the sequence shown in SEQ ID NO: 341 and VL of the sequence shown in SEQ ID NO: 353 (61) VH of the sequence shown in SEQ ID NO: 342 and VL of the sequence shown in SEQ ID NO: 351 (62) VH of the sequence shown in SEQ ID NO: 342 and VL of the sequence shown in SEQ ID NO: 352 (63) VH of the sequence shown in SEQ ID NO: 342 and VL of the sequence shown in SEQ ID NO: 353 (64) VH of the sequence shown in SEQ ID NO: 343 and VL of the sequence shown in SEQ ID NO: 354 (65) VH of the sequence shown in SEQ ID NO: 343 and VL of the sequence shown in SEQ ID NO: 355 (66) VH of the sequence shown in SEQ ID NO: 343 and VL of the sequence shown in SEQ ID NO: 356 (67) VH of the sequence shown in SEQ ID NO: 344 and VL of the sequence shown in SEQ ID NO: 354 (68) VH of the sequence shown in SEQ ID NO: 344 and VL of the sequence shown in SEQ ID NO: 355 (69) VH of the sequence shown in SEQ ID NO: 344 and VL of the sequence shown in SEQ ID NO: 356 (70) VH of the sequence shown in SEQ ID NO: 345 and VL of the sequence shown in SEQ ID NO: 354 (71) VH of the sequence shown in SEQ ID NO: 345 and VL of the sequence shown in SEQ ID NO: 355 (72) VH of the sequence shown in SEQ ID NO: 345 and VL of the sequence shown in SEQ ID NO: 356 (73) VH of the sequence shown in SEQ ID NO: 346 and VL of the sequence shown in SEQ ID NO: 354 (74) VH of the sequence shown in SEQ ID NO: 346 and VL of the sequence shown in SEQ ID NO: 355 (75) VH of the sequence shown in SEQ ID NO: 346 and VL of the sequence shown in SEQ ID NO: 356 (76) VH of the sequence shown in SEQ ID NO: 347 and VL of the sequence shown in SEQ ID NO: 357 (77) VH of the sequence shown in SEQ ID NO: 347 and VL of the sequence shown in SEQ ID NO: 358 (78) VH of the sequence shown in SEQ ID NO: 347 and VL of the sequence shown in SEQ ID NO: 359 (79) VH of the sequence shown in SEQ ID NO: 348 and VL of the sequence shown in SEQ ID NO: 357 (80) VH of the sequence shown in SEQ ID NO: 348 and VL of the sequence shown in SEQ ID NO: 358 (81) VH of the sequence shown in SEQ ID NO: 348 and VL of the sequence shown in SEQ ID NO: 359 (82) VH of the sequence shown in SEQ ID NO: 349 and VL of the sequence shown in SEQ ID NO: 357 (83) VH of the sequence shown in SEQ ID NO: 349 and VL of the sequence shown in SEQ ID NO: 358 (84) VH of the sequence shown in SEQ ID NO: 349 and VL of the sequence shown in SEQ ID NO: 359 (85) VH of the sequence shown in SEQ ID NO: 350 and VL of the sequence shown in SEQ ID NO: 357 (86) VH of the sequence shown in SEQ ID NO: 350 and VL of the sequence shown in SEQ ID NO: 358 (87) VH of the sequence shown in SEQ ID NO: 350 and VL of the sequence shown in SEQ ID NO: 359 The antibody comprises any one of the above combinations of heavy chain variable regions (VH) and light chain variable regions (VL), Optionally, the VH and VL have at least 70%, 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%, at least 99%, or 100% sequence identity with any combination of (1) to (87), and the VL has at least 70%, 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%, at least 99%, or 100% sequence identity. The antibody or antigen-binding fragment thereof that specifically binds to CDCP1 of claim 1.

3. the antibody or antigen-binding fragment thereof is selected from a Fab fragment, a Fab' fragment, a F(ab)'2 fragment, a single-chain antibody, and a disulfide-stabilized Fv protein (dsFv); The antibody or antigen-binding fragment thereof that specifically binds to CDCP1 according to claim 1 or 2, wherein the single-chain antibody is preferably selected from scFv, di-scFv, or (scFv)2.

4. The antibody or antigen-binding fragment thereof that specifically binds to CDCP1 described in any one of claims 1 to 3, wherein the antibody or antigen-binding fragment thereof is a murine antibody, a chimeric antibody, a humanized antibody, or an antibody derived from another species (e.g., rabbit, camel, or shark).

5. the antibody or antigen-binding fragment thereof further comprises a heavy chain constant region (CH) and a light chain constant region (CL); Preferably, the heavy chain constant region is selected from the heavy chain constant region of IgG, IgM, IgE, IgD or IgA or a variant thereof, and / or the light chain constant region is selected from a kappa or lambda light chain constant region or a variant thereof; The variant has one or more amino acid substitutions, deletions or additions (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions or additions, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions or additions) relative to the wild-type sequence from which it is derived, More preferably, the antibody or antigen-binding fragment thereof is (1) Human IgG1 heavy chain constant region (2) Human IgG1 light chain constant region a heavy chain constant region and a light chain constant region selected from the above, More preferably, the heavy chain constant region (CH) of the antibody or antigen-binding fragment thereof comprises the amino acid sequence set forth in SEQ ID NO: 360, and / or The antibody or antigen-binding fragment thereof that specifically binds to CDCP1 according to any one of claims 1 to 4, wherein the light chain constant region is a human IgG1 κ light chain constant region, and the light chain constant region (CL) comprises the amino acid sequence set forth in SEQ ID NO:

361.

6. the antibody or antigen-binding fragment thereof binds to CDCP1 (e.g., human CDCP1) with a KD of less than about 500 nM, e.g., less than about 100 nM, less than about 50 nM, less than about 40 nM, less than about 40 nM, less than about 20 nM, less than about 10 nM, less than about 1 nM, less than about 0.1 nM, less than about 0.01 nM or lower; and / or binds to tumor cells expressing CDCP1 with an EC50 of less than about 500 nM, e.g., less than about 100 nM, less than about 10 nM, less than about 1 nM, less than about 0.9 nM, less than about 0.8 nM, less than about 0.7 nM, less than about 0.6 nM, less than about 0.5 nM, less than about 0.4 nM, less than about 0.3 nM, less than about 0.2 nM, less than about 0.1 nM, less than about 0.01 nM or less, preferably wherein the EC50 is measured by flow cytometry or competitive ELISA; Preferably, the antibody or antigen-binding fragment thereof does not have or has endocytosis capabilities, and / or the antibody or antigen-binding fragment thereof does not have the ability to promote tumor cell migration; and / or The antibody or antigen-binding fragment thereof that specifically binds to CDCP1 according to any one of claims 1 to 5, wherein the antibody or antigen-binding fragment thereof has the ability to suppress tumor metastasis.

7. A multispecific antibody comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, A multispecific antibody, which is preferably a bispecific antibody or a trispecific antibody or a tetraspecific antibody.

8. An isolated nucleic acid molecule encoding the antibody or antigen-binding fragment thereof of any one of claims 1 to 6, or the multispecific antibody of claim 7.

9. A vector comprising the nucleic acid molecule of claim 8.

10. A host cell comprising the nucleic acid molecule of claim 8 or the vector of claim 9.

11. A method for producing an antibody or antigen-binding fragment thereof, or a multispecific antibody that specifically binds to CDCP1, comprising culturing the host cell of claim 10 under conditions that allow expression of the antibody or antigen-binding fragment thereof, or the multispecific antibody, and recovering the antibody or antigen-binding fragment thereof, or the multispecific antibody from the cultured host cell culture.

12. A complex or a pharmaceutically acceptable salt thereof, The conjugate comprises an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, or a multispecific antibody according to claim 7, and a conjugate moiety, wherein the antibody or antigen-binding fragment thereof, or the multispecific antibody is linked to the conjugate moiety directly or via a linker; Preferably, the conjugate or pharmaceutically acceptable salt thereof is characterized in that the conjugated moiety is selected from a detectable label (e.g., a radioisotope, a fluorescent substance, a luminescent substance, a colored substance, or an enzyme) or a therapeutic agent (e.g., a nuclide, an immunostimulant, an immunosuppressant, a cytokine, a toxin, and other active substances that inhibit tumor cell growth or promote tumor cell apoptosis or necrosis).

13. The structure of the complex is shown in formula (I): A-(LD)p(I), where L is present or absent; During the ceremony, A is the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, or the multispecific antibody according to claim 7; L is a linker, preferably the linker is a vc linker, the structure of which is 【Chemical 1】 and D is a conjugated moiety; Preferably, the conjugate moiety is an immunostimulant, including a TLR, STING, OND, CpG, LPS, SEB, SEA, or SEC stimulant; Preferably, the conjugate moiety is a cytokine, including IL-6, IL-15, IL-12, IL-1b, or IL-23; Preferably, the conjugate moiety is a toxin and comprises a cytotoxic agent, more preferably the cytotoxic agent is selected from camptothecins (e.g., SN-38), maytansinoids (e.g., maytansinoid DM1 / 4), pyrrolobenzodiazepines (PBD), or auristatins (e.g., Monomethyl auristatin E / F); The complex of claim 12, wherein the value of p is 0 or more, preferably p is 0 to 8.

14. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, or the multispecific antibody according to claim 7, or the conjugate according to claim 12 or 13, and a pharmaceutically acceptable carrier and / or excipient, Preferably, it further comprises another pharmaceutically active agent, More preferably, the further pharmaceutically active agent is a drug with anti-tumor activity, such as a chemotherapeutic agent (e.g., a platinum-based drug), a small molecule inhibitor, and / or the additional pharmaceutically active agent is an immunotherapeutic agent, such as an immune checkpoint inhibitor (e.g., a PD-1, PD-L1, or CTLA-4 inhibitor), an immune enhancing agent (e.g., an interferon, an interleukin), or an immune activator (e.g., a TLR, OND, CpG, LPS, SEB, SEA, STING, or SEC activator); and / or the additional pharmaceutically active agent is an oncolytic virus, an immune cell, or an engineered immune cell; and / or A pharmaceutical composition wherein said additional pharmaceutically active agent is a radioactive substance or a color-producing agent.

15. A kit comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, or the multispecific antibody according to claim 7, or the conjugate according to claim 12 or 13, or the pharmaceutical composition according to claim 14.

16. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, or the multispecific antibody according to claim 7, or the conjugate according to claim 12 or 13, or the pharmaceutical composition according to claim 14, for the manufacture of a medicament for the prevention and / or treatment and / or adjuvant therapy and / or neoadjuvant therapy of a CDCP1-associated disease.

17. the CDCP1-associated disease is a tumor; Preferably, the tumor is selected from a solid tumor or a hematological tumor; More preferably, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

18. the antibody or antigen-binding fragment thereof, the conjugate, the multispecific antibody or the pharmaceutical composition is administered separately, in combination, simultaneously or sequentially with another pharmaceutically active agent; Preferably, the other pharmaceutically active agent is a drug with antitumor activity (e.g., a platinum-based drug, a small molecule inhibitor), or the other pharmaceutically active agent is an immune-enhancing drug (e.g., an interferon, an interleukin), or the other pharmaceutically active agent is an immune-stimulating agent (e.g., a TLR, OND, CpG, LPS, SEB, SEA, STING, or SEC activator), or the other pharmaceutically active agent is an immune checkpoint inhibitor (e.g., a PD-1, PD-L1, or CTLA-4 inhibitor), or the other pharmaceutically active agent is an oncolytic virus, an immune cell, or an engineered immune cell.

19. A method for the prevention and / or treatment and / or adjuvant therapy and / or neoadjuvant therapy of a CDCP1-associated disease in a subject, comprising administering to a subject in need thereof an effective amount of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, or a multispecific antibody according to claim 7, or a conjugate according to claim 12 or 13, or a pharmaceutical composition according to claim 14.

20. further comprising administering to the subject a second therapy, wherein the second therapy is selected from surgery, chemotherapy, radiation therapy, immunotherapy, gene therapy, DNA therapy, RNA therapy, nanotherapy, viral therapy, adjuvant therapy, cell therapy, and any combination thereof; 20. The method of claim 19, wherein optionally the second therapy may be used separately or in combination with the method of claim 19.

21. the CDCP1-associated disease is a tumor, preferably the tumor is selected from a solid tumor or a hematological tumor; More preferably, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

22. A method for detecting the presence or level of CDCP1 in a sample, comprising contacting the sample with the antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, the multispecific antibody according to claim 7, the conjugate according to claim 12 or 13, or the pharmaceutical composition according to claim 14 under conditions allowing the formation of a complex between CDCP1 and the antibody or antigen-binding fragment thereof, the multispecific antibody, the conjugate, or the pharmaceutical composition, and detecting the formation of the complex.

23. Use of the antibody or antigen-binding fragment according to any one of claims 1 to 6, or the multispecific antibody according to claim 7, or the conjugate according to claim 12 or 13, or the pharmaceutical composition according to claim 14, for the manufacture of a diagnostic kit, comprising: The kit is used for the diagnosis or differential diagnosis of a CDCP1-associated disease, and preferably, the CDCP1-associated disease is a tumor, and more preferably, the tumor is selected from a solid tumor or a blood tumor, and even more preferably, the solid tumor is selected from esophageal cancer, digestive cancer, pancreatic cancer, thyroid cancer, colorectal cancer, kidney cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, gastric cancer, esophagogastric junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, or glioblastoma, and the blood tumor is selected from lymphoma, myeloma (e.g., multiple myeloma), or leukemia.

24. A CDCP1-associated disease, preferably the CDCP1-associated disease is a tumor, more preferably the tumor is selected from a solid tumor or a hematological tumor; More preferably, the solid tumor is selected from esophageal cancer, gastrointestinal cancer, pancreatic cancer, thyroid cancer, colorectal cancer, renal cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma or small cell lung cancer), liver cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma or glioblastoma, and the hematological tumor is selected from lymphoma, myeloma (e.g., multiple myeloma) or leukemia,

25. The CDCP1-associated disease is preferably a tumor, more preferably the tumor is selected from solid tumors and hematological tumors, and even more preferably the solid tumor is selected from esophageal cancer, digestive tract cancer, pancreatic cancer, thyroid cancer, colorectal cancer, kidney cancer, lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, or small cell lung cancer), liver cancer, stomach cancer, gastroesophageal junction (GEJ) adenocarcinoma, head and neck cancer, bladder cancer, breast cancer, uterine cancer, cervical cancer, and ovarian cancer.

15. The antibody or antigen-binding fragment of any one of claims 1 to 6, or the multispecific antibody of claim 7, or the conjugate of claim 12 or 13, or the pharmaceutical composition of claim 14, for use in the diagnosis or differential diagnosis of a disease selected from the group consisting of cancer, prostate cancer, testicular cancer, germ cell tumor, bone tumor, skin cancer, thymic cancer, bile duct cancer, gallbladder cancer, melanoma, mesothelioma, sarcoma, and glioblastoma, and the hematological tumor is selected from the group consisting of lymphoma, myeloma (e.g., multiple myeloma), and leukemia.

Images