Anti-tumor-associated antigen antibodies and uses thereof

Abstract

Anti-TAA antibodies and antigen-binding fragments thereof are described, as are nucleic acids encoding the antibodies, compositions comprising the antibodies, and methods of producing the antibodies and methods of using the antibodies to treat or prevent diseases, such as cancer and / or inflammatory diseases.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 021,215, filed May 7, 2020; U.S. Provisional Patent Application No. 62 / 706,131, filed August 3, 2020; U.S. Provisional Patent Application No. 63 / 198,420, filed October 16, 2020; and U.S. Patent Application No. 63 / 131,394, filed December 29, 2020, the disclosures of each of which are incorporated herein by reference in their entirety.

[0002] FIELD OF THE INVENTION The present invention relates to monoclonal anti-tumor-associated antigen (TAA) antibodies, nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. Methods of making the antibodies and methods of using the antibodies to treat diseases, including cancer and inflammatory diseases and / or related complications, are also provided.

[0003] Reference to an electronically submitted sequence listing This application contains a Sequence Listing that has been submitted electronically via EFS-Web as a Sequence Listing in ASCII format having the file name "065799.34WO1 Sequence Listing" and a creation date of April 28, 2021, with a size of 150 kb. The Sequence Listing submitted via EFS-Web is a part of the specification and is hereby incorporated by reference in its entirety. [Background technology]

[0004] "Tumor-associated antigen (TAA)" refers to any cell surface peptide and / or antigen, or combination of cell surface peptides and / or antigens, and post-translational modifications (such as glycosylation) that are present at higher levels in tumors than in normal tissues. Some tumor-associated antigens that are specifically present in tumors are also known as tumor-specific antigens (TSAs). Examples of tumor-associated antigens include viral proteins encoded by oncogenic viruses; mutated oncoproteins or tumor suppressors; normal proteins that are overexpressed on and / or in tumor cells; post-translational modifications of cell surface proteins; oncofetal proteins whose expression is usually restricted during development but not in adult tissues; and cell type-specific proteins whose expression is restricted to nonessential tissues.

[0005] Tax-interacting protein 1 (TIP-1, also known as Tax1bp3 or glutaminase-interacting protein, GIP) is a PDZ (PSD-95 / Discs large / ZO-1 homology) domain-containing intracellular protein (124 amino acids in humans and mice). A single PDZ domain encompassing residues 13 to 112 of the 124-amino acid protein (Olalla et al., FEBS Lett 2001;488:116-122) is the only functional and structural unit that can be identified in TIP-1, suggesting that TIP-1 is unique among PDZ-containing proteins and may perform its function only through protein-protein interactions. TIP-1 interacts with numerous intracellular proteins via its PDZ domain, including glutaminase L, β-catenin, FAS, HTLV Tax, HPV E6, Rhotekin, and Kir 2.3 (Zoetewey et al., Biochemistry 2011; 50:3528-3539). However, the precise biological function of TIP-1 remains unclear. Elevated TIP-1 expression has been detected in human invasive breast cancer cells and has been shown to be associated with tumor cell adhesion, migration, and lung metastasis (Han et al., Biochem Biophys Res Commun 2012; 422:139-145). Recently, TIP-1 was found to translocate to the surface of cancer cells upon irradiation induction (Yan et al., Oncotarget 2016; 7:43352-43362), suggesting that TIP-1 may be a tumor neoantigen upon induction. Therefore, anti-TIP-1 monoclonal antibodies (mAbs) can be used to target cancer cells with selectivity and serve as potential therapeutic agents.

[0006] Glypican-3 (GPC3) is a GPI-anchored cell surface glycoprotein containing a 66 kDa core protein and two heparan sulfate (HS) glycosaminoglycan polysaccharide chains. It belongs to the glypican family, which has six members (GPC1-GPC6). GPC3 is an oncofetal protein that is expressed only during embryonic development, but is overexpressed in more than 70% of hepatocellular carcinomas (HCC) (Baumhoer, D. et al., Am J Clin Pathol. 2008 Jun;129(6):899-906). Its restricted expression in cancer cells makes GPC3 a tumor-specific antigen (TSA) that can be exploited to target cancer cells with monoclonal antibody-based therapy. In addition to its tumor-specific expression, GPC3 may also promote tumor growth. GPC3 can regulate the Wnt signaling pathway. The core protein of GPC3 interacts with Wnt, functions as a coreceptor, and activates frizzled (FZD) and downstream signaling via β-catenin and YAP (Capurro, MI et al., Cancer Res. 2005 Jul 15; 65(14):6245-54). In fact, Wnt signaling is often activated in HCC (Khalaf, AM., J Hepatocell Carcinoma. 2018; 5:61-73). Therefore, GPC3 is a promising therapeutic target for treating HCC and other cancers that overexpress GPC3. Summary of the Invention

[0007] In one general aspect, the invention relates to isolated monoclonal antibodies or antigen-binding fragments thereof that specifically bind to TAAs, such as TIP-1 and GPC3.

[0008] (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively; and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequences shown below. The antibody or antigen-binding fragment thereof specifically binds to GPC3, preferably human GPC3.

[0009] (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55 and 56, respectively; (6) SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; or (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively; and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequences shown below. The antibody or antigen-binding fragment thereof specifically binds to GPC3, preferably human GPC3.

[0010] (1) SEQ ID NOs: 3, 4, 5, 6, 7 and 8, respectively An isolated monoclonal antibody or antigen-binding fragment thereof is provided, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence:

[0011] (1) SEQ ID NOs: 9, 10, 11, 12, 13 and 14, respectively An isolated monoclonal antibody or antigen-binding fragment thereof is provided, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence:

[0012] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, 71, or 1, or a light chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, 72, or 2.

[0013] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (b) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; (c) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (d) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (e) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (f) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (g) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; or (h) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2 Includes:

[0014] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof binds to a TAA and is capable of inducing effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or mediating the recruitment of a conjugated drug, and / or forming a bispecific antibody with another monoclonal antibody or antigen-binding fragment thereof that has cancer-killing activity.

[0015] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is chimeric.

[0016] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is human or humanized. In certain embodiments, the humanized monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 99-102, 120-123, 126-132, 139-140, 143-149, 153-156, 160-163, 183-197, or 202-205, or a light chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 103-105, 124-125, 133-138, 141-142, 150-152, 157-159, 164-166, or 198-201.

[0017] In certain embodiments, the humanized monoclonal antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 Includes:

[0018] Also provided is an isolated bispecific antibody comprising the monoclonal antibody of the invention or an antigen-binding fragment thereof.

[0019] Also provided is an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody of the invention.

[0020] Also provided is a vector comprising an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the invention.

[0021] Host cells comprising vectors containing isolated nucleic acids encoding the monoclonal antibodies or antigen-binding fragments thereof, or bispecific antibodies or antigen-binding fragments thereof of the invention are also provided.

[0022] In certain embodiments, a pharmaceutical composition is provided comprising an isolated monoclonal antibody or antigen-binding fragment thereof, or an isolated bispecific antibody or antigen-binding fragment thereof of the invention and a pharmaceutically acceptable carrier.

[0023] Also provided is a method for specifically targeting a TAA (e.g., TIP-1 or GPC3) on the surface of a cancer cell in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the present invention.

[0024] Also provided is a method for treating cancer in a subject in need thereof, comprising administering the pharmaceutical composition of the present invention to the subject. The cancer may be any liquid or solid cancer, for example, but not limited to, lung cancer, gastric cancer, esophageal cancer, bile duct cancer, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors.

[0025] Also provided is a method of treating an inflammatory disease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the present invention.

[0026] Also provided is a method for producing a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the present invention, comprising culturing cells containing nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, under conditions to produce the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, and recovering the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof from the cells or culture.

[0027] Also provided is a method for producing a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising combining the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

[0028] Also provided is a method for determining the level of a TAA (e.g., TIP-1 or GPC3) in a subject. The method includes (a) obtaining a sample from the subject, (b) contacting the sample with an antibody or antigen-binding fragment thereof of the present invention, and (c) determining the level of the TAA in the subject. In certain embodiments, the sample is a tissue sample. The tissue sample can be, for example, a cancer tissue sample. In certain embodiments, the sample is a blood sample.

[0029] In another general aspect, the invention relates to a chimeric antigen receptor (CAR) construct that induces T cell-mediated cancer killing, the CAR construct comprising at least one antigen-binding domain that specifically binds human GPC3, a hinge region, a transmembrane region, and an intracellular signaling domain.

[0030] An isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR) is provided. The CAR can comprise: (a) an extracellular domain comprising at least one antigen-binding domain that specifically binds to GPC3; (b) a hinge region; (c) a transmembrane region; and (d) an intracellular signaling domain.

[0031] In certain embodiments, the antigen binding domain comprises: (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively; and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence shown below, and the antigen-binding domain specifically binds to GPC3, preferably human GPC3.

[0032] In certain embodiments, the antigen binding domain comprises: (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55 and 56, respectively; (6) SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; or (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively; and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence shown below, and the antigen-binding domain specifically binds to GPC3, preferably human GPC3.

[0033] In certain embodiments, the antigen-binding domain comprises a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, or 71, or a light chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, or 72.

[0034] In certain embodiments, the antigen binding domain comprises: a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; or g. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72 Includes:

[0035] In certain embodiments, the antigen-binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 99-102, 120-123, 126-132, 139-140, 143-149, 153-156, 160-163, 183-197, or 202-205, or a light chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 103-105, 124-125, 133-138, 141-142, 150-152, 157-159, 164-166, or 198-201.

[0036] In certain embodiments, the antigen-binding domain is humanized, (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 Includes:

[0037] In certain embodiments, the antigen-binding domain is a single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3.

[0038] In certain embodiments, the antigen-binding domain is a humanized single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3. In certain embodiments, the humanized single-chain variable fragment (scFv) comprises a polypeptide sequence at least 95% identical to any one of SEQ ID NOs: 167-182.

[0039] In certain embodiments, a chimeric antigen receptor (CAR) comprises one or more antigen-binding domains.

[0040] In certain embodiments, the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0041] Also provided is a chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of the invention.

[0042] Also provided is a vector comprising an isolated polynucleotide comprising a nucleic acid encoding a CAR of the invention.

[0043] Host cells comprising the vectors of the invention are also provided.

[0044] In certain embodiments, the host cell is a T cell, preferably a human T cell. In certain embodiments, the host cell is a NK cell, preferably a human NK cell. The T cell or NK cell can be engineered to express a CAR of the present invention to treat a disease such as, for example, cancer.

[0045] Also provided is a method of making a host cell expressing a chimeric antigen receptor (CAR) of the invention, the method comprising transducing a T cell or NK cell with a vector comprising an isolated nucleic acid encoding a CAR of the invention.

[0046] Also provided is a method for producing CAR-T cells or CAR-NK cells of the invention, comprising culturing T cells or NK cells containing an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) of the invention under CAR-T cell or CAR-NK cell-producing conditions, and recovering the CAR-T cells or CAR-NK cells.

[0047] Also provided is a method for generating a population of RNA engineered cells comprising a chimeric antigen receptor (CAR) of the invention, the method comprising contacting the cells with an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) of the invention, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

[0048] Also provided are methods of treating cancer in a subject in need thereof, comprising administering to the subject the CAR-T cells and / or CAR-NK cells of the invention. The cancer can be any liquid or solid cancer, for example, it can be selected from, but is not limited to, lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors.

[0049] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that increases the efficacy of cells expressing a CAR molecule.

[0050] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that ameliorates one or more side effects associated with administration of cells expressing a CAR molecule.

[0051] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that treats a GPC3-associated disease.

[0052] The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the appended drawings. It should be understood, however, that the application is not limited to the precise embodiments shown in the drawings. The present invention also relates to the following: [Item 1] (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively; an isolated monoclonal antibody or an antigen-binding fragment thereof, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequences shown below, wherein the antibody or antigen-binding fragment thereof specifically binds to GPC3, preferably human GPC3. [Item 2] (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55 and 56, respectively; (6) SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; or (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively; an isolated monoclonal antibody or an antigen-binding fragment thereof, comprising a heavy chain complementarity-determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity-determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequences shown below, wherein the antibody or antigen-binding fragment thereof specifically binds to GPC3, preferably human GPC3. [Item 3] (1) SEQ ID NOs: 3, 4, 5, 6, 7 and 8, respectively An isolated monoclonal antibody or an antigen-binding fragment thereof, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of: [Item 4] (1) SEQ ID NOs: 9, 10, 11, 12, 13 and 14, respectively An isolated monoclonal antibody or an antigen-binding fragment thereof, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of: [Item 5] 5. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 4, comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, 71, or 1, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, 72, or 2. [Item 6] a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; g. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; or h. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2 6. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 5, comprising: [Item 7] 7. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 6, wherein the antibody or antigen-binding fragment thereof is chimeric and / or human or humanized. [Item 8] a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 99 to 102, 120 to 123, 126 to 132, 139 to 140, 143 to 149, 153 to 156, 160 to 163, 183 to 197, or 202 to 205; or 8. The isolated monoclonal antibody or antigen-binding fragment thereof according to Item 7, comprising a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 103 to 105, 124 to 125, 133 to 138, 141 to 142, 150 to 152, 157 to 159, 164 to 166, or 198 to 201. [Item 9] The humanized monoclonal antibody or antigen-binding fragment thereof is (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 9. The isolated monoclonal antibody or antigen-binding fragment thereof according to item 8, comprising: [Item 10] 10. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 9, wherein the antibody or antigen-binding fragment thereof is capable of inducing effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or mediating the recruitment of a conjugated drug, and / or forming a bispecific antibody with another mAb or antigen-binding fragment thereof that has cancericidal activity. [Item 11] A bispecific antibody or an antigen-binding fragment thereof, comprising the monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 10. [Item 12] 11. An isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 10. [Item 13] 12. An isolated nucleic acid encoding the bispecific antibody or antigen-binding fragment thereof according to item 11. [Item 14] 14. A vector comprising the isolated nucleic acid of item 12 or 13. [Item 15] A host cell containing the vector according to item 14. [Item 16] A pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 10, or the bispecific antibody or antigen-binding fragment thereof according to item 11, and a pharmaceutically acceptable carrier. [Item 17] 17. A method for targeting a TAA on the surface of cancer cells and / or treating cancer and / or treating an inflammatory disease in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of Item 16, wherein optionally the cancer is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors. [Item 18] A method for producing the monoclonal antibody or antigen-binding fragment thereof according to any one of Items 1 to 10, or the bispecific antibody or antigen-binding fragment thereof according to Item 11, the method comprising culturing cells containing a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, under conditions for producing the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, and recovering the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof from the cells or culture. [Item 19] A method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment thereof according to any one of Items 1 to 10, or the bispecific antibody or antigen-binding fragment thereof according to Item 11, the method comprising combining the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain a pharmaceutical composition. [Item 20] 1. A method for determining the level of a TAA in a subject, comprising: a. obtaining a sample from a subject; b. contacting the sample with the isolated monoclonal antibody or antigen-binding fragment thereof according to any one of items 1 to 10; and c. Determining the level of TAA in a subject A method comprising: [Item 21] 21. The method according to item 20, wherein the sample is a tissue sample or a blood sample, and optionally the tissue sample is a cancer tissue sample. [Item 22] 1. An isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an extracellular domain comprising at least one antigen-binding domain that specifically binds to GPC3; (b) hinge region; (c) a transmembrane region, and (d) Intracellular signaling domain An isolated polynucleotide comprising: [Item 23] The antigen-binding domain is (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively 23. The isolated polynucleotide according to Item 22, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence: [Item 24] The antigen-binding domain is (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; or (6) SEQ ID NOs: 65, 66, 67, 68, 69 and 70, respectively; (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively 23. The isolated polynucleotide according to Item 22, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence: [Item 25] 25. The isolated polynucleotide of any one of Items 22 to 24, wherein the antigen-binding domain comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, or 71, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, or 72. [Item 26] The antigen-binding domain is a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; or g. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72 26. The isolated polynucleotide of item 25, comprising: [Item 27] 25. The isolated polynucleotide of any one of Aspects 22 to 24, wherein the antigen-binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 99 to 102, 120 to 123, 126 to 132, 139 to 140, 143 to 149, 153 to 156, 160 to 163, 183 to 197, or 202 to 205, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 103 to 105, 124 to 125, 133 to 138, 141 to 142, 150 to 152, 157 to 159, 164 to 166, or 198 to 201. [Item 28] The antigen-binding domain is humanized, (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 28. The isolated polynucleotide of Item 27, comprising: [Item 29] 29. The isolated polynucleotide according to any one of Items 22 to 28, wherein the antigen-binding domain is a single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3. [Item 30] 30. The isolated polynucleotide of item 29, wherein the single-chain variable fragment (scFv) is humanized. [Item 31] 31. The isolated polynucleotide of item 29 or 30, wherein the single-chain variable fragment (scFv) comprises a polypeptide sequence at least 95% identical to any one of SEQ ID NOs: 167 to 182. [Item 32] 32. The isolated polynucleotide of any one of items 22 to 31, wherein the chimeric antigen receptor (CAR) comprises one or more antigen-binding domains. [Item 33] 33. The isolated polynucleotide of any one of items 22 to 32, wherein the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains. [Item 34] A chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of any one of items 22 to 33. [Item 35] A vector comprising the isolated polynucleotide according to any one of Items 22 to 33. [Item 36] A host cell containing the vector described in item 35. [Item 37] 37. The host cell according to item 36, wherein the host cell is a T cell, preferably a human T cell. [Item 38] 37. The host cell according to item 36, wherein the host cell is a NK cell, preferably a human NK cell. [Item 39] 36. A method for producing a host cell expressing a chimeric antigen receptor (CAR), comprising transducing a T cell with the vector of item 35. [Item 40] 34. A method for producing chimeric antigen receptor (CAR)-T cells, the method comprising culturing T cells containing an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of Items 22 to 33 under CAR-T cell-producing conditions, and recovering the CAR-T cells. [Item 41] 36. A method for producing a host cell expressing a chimeric antigen receptor (CAR), comprising transducing an NK cell with the vector of item 35. [Item 42] 34. A method for producing chimeric antigen receptor (CAR)-NK cells, the method comprising culturing NK cells containing an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of Items 22 to 33 under CAR-NK cell-producing conditions, and recovering the CAR-NK cells. [Item 43] 34. A method for generating a cell comprising a chimeric antigen receptor (CAR), the method comprising contacting a cell with an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of items 22 to 33, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA. [Item 44] A method for treating cancer in a subject in need thereof, comprising administering to the subject the host cell of any one of Items 36 to 38, wherein optionally the cancer is selected from lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors. [Item 45] 45. The method of item 44, further comprising administering to a subject in need thereof an agent that increases the efficacy of the CAR-expressing cell. [Item 46] 45. The method of item 44, further comprising administering to a subject in need thereof an agent that ameliorates one or more side effects associated with administration of the cells expressing a CAR. [Item 47] Item 45. The method of item 44, further comprising administering to a subject in need thereof a drug for treating a GPC3-associated disease. [Brief explanation of the drawings]

[0053] [Figure 1] Figure 1 shows the binding of purified chimeric anti-TIP-1 mAb T5C (the VH and VL regions of murine mAb T5 fused to the constant regions of human IgG1 heavy and kappa light chains, respectively) to immobilized TIP-1 in an ELISA assay. The antigen was human TIP-1 tagged with Myc at the C-terminus (Origene, CAT#: TP304776). [Figure 2A] Figures 2A-2B show the binding of purified chimeric anti-GPC3 mAb (VH and VL regions of mouse anti-GPC3 mAb fused to the constant regions of human IgG1 heavy and kappa light chains, respectively) to GPC3. Figures 2A-2B show the binding of anti-GPC3 mAb to immobilized human GPC3 [FLAG-huGPC3-His; a human GPC3 (25-559) fusion protein with a FLAG tag at the N-terminus and a His tag at the C-terminus] in an ELISA assay. Figures 2C-2E show the binding of anti-GPC3 mAb to a HEK293 stable cell line expressing human GPC3 using FACS analysis. Figures 2F-2G show the binding of anti-GPC3 mAb at 26 nM to control HEK293 cells (without exogenously introduced GPC3 expression) using FACS analysis. Isotype control: negative control antibody; secondary antibody only: only secondary antibody was added to the assay as a negative control. [Figure 2B] Continuation of Figure 2A. [Figure 2C] Continuation of Figure 2B. [Figure 2D] This is a continuation of Figure 2C. [Figure 2E] This is a continuation of Figure 2D. [Figure 2F] Continuation of Figure 2E. [Figure 2G] Continuation of Figure 2F. [Figure 3A] Figure 1 shows the binding of humanized anti-GPC3 mAbs to a HEK293 stable cell line expressing human GPC3 using FACS analysis. Isotype control, negative control antibody; secondary antibody only, secondary antibody alone was added to the assay as a negative control. M3-C, chimeric version of M3; the same naming convention is used for 26A1, 36F8, 39E1, 43B9, and F7. [Figure 3B] This is a continuation of Figure 3A. [Figure 3C] This is a continuation of Figure 3B. [Figure 3D] This is a continuation of Figure 3C. [Figure 3E] This is a continuation of Figure 3D. [Figure 3F] Continuation of Figure 3E. [Figure 3G] Continuation of Figure 3F. [Figure 3H] Continuation of Figure 3G. [Figure 3I] This is a continuation of Figure 3H. [Figure 3J] Continuation of Figure 3I. [Figure 3K] This is a continuation of Figure 3J. [Figure 3L] Continuation of Figure 3K. [Figure 3M] This is a continuation of Figure 3L. [Figure 3N] Continuation of Figure 3M. [Figure 3O] This is a continuation of Figure 3N. [Figure 4A] Figure 1 shows the binding of anti-GPC3 scFv molecules to a HEK293 stable cell line expressing human GPC3 using FACS analysis. Isotype control, negative control antibody; secondary antibody only, secondary antibody only was added to the assay as a negative control. [Figure 4B] This is a continuation of Figure 4A. [Figure 4C] This is a continuation of Figure 4B. [Figure 4D] This is a continuation of Figure 4C. [Figure 4E] This is a continuation of Figure 4D. DETAILED DESCRIPTION OF THE INVENTION

[0054] Throughout the Background and specification, various publications, articles, and patents are cited or described. Each of these references is incorporated herein by reference in its entirety. The discussion of documents, acts, materials, devices, articles and the like which has been included in the specification is for the purpose of providing a context for the present invention. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any invention(s) disclosed or claimed.

[0055] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Unless otherwise defined, certain terms used herein have the meaning given in the specification.

[0056] It must be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0057] Unless otherwise specified, any numerical values, e.g., concentrations or concentration ranges, described herein should be understood to be modified in all instances by the term "about." Thus, numerical values ​​generally include ±10% of the recited value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, unless the context clearly dictates otherwise, the use of numerical ranges explicitly includes all individual numerical values ​​within that range, including all possible subranges, integers, and fractions of values ​​within such ranges.

[0058] Unless otherwise indicated, the term "at least" before a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0059] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains," or "containing," or any other variation thereof, are understood to imply that the specified integer or group of integers is inclusive, but not the exclusion of any other integer or group of integers, and are non-exclusive or open-ended. For example, a composition, mixture, process, method, article, or device containing a list of elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent in such composition, mixture, process, method, article, or device. Furthermore, unless expressly stated otherwise, "or" refers to an inclusive or, not an exclusive or. For example, condition A or B is satisfied by any one of the following: A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), and both A and B are true (or exist).

[0060] As used herein, the conjunction "and / or" between multiple listed elements is understood to encompass the individual and combined options. For example, when two elements are joined by "and / or," the first option refers to the application of the first element without the second element. The second option refers to the application of the second element without the first element. The third option refers to the application of the first and second elements together. Any one of these options is understood to fall within this meaning and therefore meet the requirements of the term "and / or" as used herein. The simultaneous application of more than one of these options is also understood to fall within this meaning and therefore meet the requirements of the term "and / or."

[0061] As used herein, the term "consists of" or variations such as "consist of" or "consisting of," as used throughout the specification and claims, indicates that any recited integer or group of integers is inclusive, but that no additional integers or groups of integers can be added to the specified method, structure, or composition.

[0062] As used herein, the term "consists essentially of" or variations such as "consist essentially of" or "consisting essentially of" indicates the inclusion of any recited integer or group of integers, and, where appropriate, the inclusion of any recited integer or group of integers that does not materially alter the basic or novel characteristics of the specified method, structure, or composition, as used throughout the specification and claims. See MPEP § 2111.03.

[0063] As used herein, "subject" refers to any animal, preferably a mammal, and most preferably a human. As used herein, the term "mammal" encompasses any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., more preferably humans.

[0064] The words "right", "left", "bottom" and "top" designate directions in the figures to which reference is made.

[0065] It should also be understood that the terms "about," "approximately," "generally," "substantially," and similar terms used herein when referring to dimensions or characteristics of preferred inventive components indicate that the described dimensions / characteristics are not precise boundaries or parameters, as understood by those skilled in the art, and do not exclude minor variations therefrom that are functionally the same or similar. At a minimum, such references involving numerical parameters are expected to include variations that are not expected to alter the lowest numerical value, using mathematical and industrial principles accepted in the art (e.g., rounding, measurement or other systematic errors, manufacturing tolerances, etc.).

[0066] The term "identical" or percent "identity," in the context of two or more nucleic acid or polypeptide sequences (e.g., anti-TAA antibodies and the polynucleotides that encode them, TAA polypeptides and the TAA polynucleotides that encode them, chimeric antigen receptors (CARs) comprising an antigen-binding domain specific for GPC3 and the polynucleotides that encode them), refers to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when aligned compared to maximum correspondence as determined using one of the following sequence comparison algorithms or by visual inspection:

[0067] For sequence comparison, typically one sequence serves as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence, based on the designated program parameters.

[0068] Optimal alignment of sequences for comparison can be determined, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 1981; 2:482, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 1970; 48:443, by the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 1988; 85:2444, by computer implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (generally, Current Protocols in Molecular Biology, F.M. Ausubel et al., eds., Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., 1995). (See Supplement (Ausubel)).

[0069] Examples of suitable algorithms for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., J. Mol. Biol. 1990; 215: 403-410 and Altschul et al., Nucleic Acids Res. 1997; 25: 3389-3402, respectively. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that, when aligned with words of the same length in a database sequence, match or meet some positive threshold score T. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased.

[0070] Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is ​​used to calculate the cumulative score. Extension of word hits in each direction is halted if: the cumulative alignment score falls by an amount X from its maximum achieved value; the accumulation of one or more negative-scoring residue alignments causes the cumulative score to fall below zero; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 1989;89:10915).

[0071] In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA 1993; 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered to be similar to a reference sequence if the smallest sum probability in the comparison of the test nucleic acid with the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

[0072] A further indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the polypeptide encoded by the second nucleic acid, as described below. Thus, for example, a polypeptide will generally be substantially identical to a second polypeptide where the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions.

[0073] As used herein, the term "isolated" indicates that a biological component (e.g., a nucleic acid, peptide, or protein) has been substantially separated, produced separately, or purified from other biological components of the organism in which it naturally occurs, i.e., from other chromosomal and extrachromosomal DNA and RNA, and proteins. Thus, "isolated" nucleic acids, peptides, and proteins include nucleic acids and proteins purified by standard purification methods. "Isolated" nucleic acids, peptides, and proteins can be part of a composition and still be isolated, if the composition is not part of the nucleic acid, peptide, or protein's natural environment. The term also encompasses nucleic acids, peptides, and proteins prepared by recombinant expression in a host cell, as well as chemically synthesized nucleic acids.

[0074] As used herein, the term "polynucleotide," which may be interchangeably referred to as "nucleic acid molecule," "nucleotide," or "nucleic acid," refers to any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. "Polynucleotide" includes, but is not limited to, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is a mixture of single- and double-stranded regions, and hybrid molecules containing DNA and RNA that may be single-stranded or, more commonly, double-stranded, or a mixture of single- and double-stranded regions. Furthermore, "polynucleotide" refers to triple-stranded regions containing RNA or DNA, or both RNA and DNA. The term polynucleotide also includes DNA or RNA containing one or more modified bases and DNA or RNA with backbones modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and unusual bases, such as inosine. Various modifications can be made to DNA and RNA. Thus, "polynucleotide" encompasses chemically, enzymatically, or metabolically modified forms of polynucleotides commonly found in nature, as well as the DNA and RNA chemical forms characteristic of viruses and cells. "Polynucleotide" also encompasses relatively short nucleic acid strands often referred to as oligonucleotides.

[0075] As used herein, the term "vector" is a replicon into which another nucleic acid segment may be operatively inserted so as to bring about the replication or expression of the segment.

[0076] As used herein, the term "host cell" refers to a cell that contains a nucleic acid molecule of the invention. A "host cell" can be any cell type, e.g., a primary cell, a cultured cell, or a cell from a cell line. In one embodiment, a "host cell" is a cell that has been transfected with a nucleic acid molecule of the invention. In another embodiment, a "host cell" is the progeny or potential progeny of such a transfected cell. The progeny of a cell may or may not be identical to the parent cell due to, for example, mutations or environmental influences that may occur in subsequent generations, or due to integration of the nucleic acid molecule into the host cell genome.

[0077] As used herein, the term "expression" refers to the biosynthesis of a gene product. The term includes transcription of a gene into RNA. The term also includes translation of RNA into one or more polypeptides, and further includes all naturally occurring post-transcriptional and post-translational modifications. The expressed antibody may be in the cytoplasm of a host cell, in an extracellular environment, such as the growth medium of a cell culture, or anchored to the cell membrane.

[0078] As used herein, the terms "peptide," "polypeptide," or "protein" can refer to a molecule composed of amino acids and recognized as a protein by those skilled in the art. Conventional one-letter or three-letter codes for amino acid residues are used herein. The terms "peptide," "polypeptide," and "protein" can be used interchangeably herein to refer to polymers of amino acids of any length. The polymers can be linear or branched, can contain modified amino acids, and can be interrupted by non-amino acids. The terms also encompass amino acid polymers that are modified naturally or by intervention, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids), as well as other modifications known in the art.

[0079] The peptide sequences described herein are written according to the usual convention whereby the N-terminal region of the peptide is on the left and the C-terminal region is on the right. Although isomeric forms of amino acids are known, unless expressly indicated otherwise, the amino acids represented are in the L-form.

[0080] Chimeric antigen receptor (CAR) As used herein, the term "chimeric antigen receptor" (CAR) refers to a recombinant polypeptide that includes at least an extracellular domain that specifically binds to an antigen or target, a transmembrane domain, and an intracellular T cell receptor activation signaling domain. When the extracellular domain of the CAR binds to a target antigen on the surface of a target cell, clustering of the CAR occurs, providing an activation stimulus to the cell containing the CAR. CARs redirect the specificity of immune effector cells, triggering proliferation, cytokine production, phagocytosis, and / or production of molecules that can mediate cell death of target antigen-expressing cells in a major histocompatibility (MHC)-independent manner.

[0081] In one embodiment, a CAR comprises an antigen-binding domain, a hinge region, a costimulatory domain, an activation domain, and a transmembrane region. In one embodiment, a CAR comprises an antigen-binding domain, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region. In one embodiment, a CAR comprises two antigen-binding domains, a hinge region, a costimulatory domain, an activation domain, and a transmembrane region. In one embodiment, a CAR comprises two antigen-binding domains, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region.

[0082] As used herein, the term "signal peptide" refers to a leader sequence at the amino-terminus (N-terminus) of a nascent CAR protein that co- or post-translationally targets the nascent protein to the endoplasmic reticulum, leading to subsequent surface expression.

[0083] As used herein, the term "extracellular antigen-binding domain," "extracellular domain," or "extracellular ligand-binding domain" refers to the portion of a CAR that is located on the outside of the cell membrane and is capable of binding to an antigen, target, or ligand.

[0084] As used herein, the term "hinge region" refers to the portion of a CAR that connects two adjacent domains of a CAR protein, e.g., the extracellular domain and the transmembrane domain.

[0085] As used herein, the term "transmembrane domain" refers to the portion of a CAR that extends across the cell membrane and anchors the CAR to the cell membrane. This is sometimes referred to as the "transmembrane region."

[0086] Costimulatory domain As used herein, chimeric antigen receptors can incorporate a costimulatory (signaling) domain to enhance their efficacy. The costimulatory (signaling) domain can be derived from a costimulatory molecule. A costimulatory molecule is a cell surface molecule other than an antigen receptor or its ligand that is required for an efficient immune response.The costimulatory domain can be derived from costimulatory molecules, including CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (alpha, beta, delta, epsilon, gamma, zeta), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, programmed death-1 (PD-1), inducible T cell costimulatory factor (ICOS), lymphocyte function-associated antigen-1 (LFA-1; CD11a and CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Ig alpha (CD79a), DAP10, Fc gamma receptor, MHC class I molecule, TNFR, integrin, signaling lymphocyte activation molecule, BTLA, Toll ligand receptor, ICAM-1, CDS, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD8 alpha, CD8 beta, IL-2R beta, IL-2R gamma, IL-7R alpha , ITGA4, VLA1, CD49a, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, ITGAE, CD103, ITGAL, CD1a, CD1b, CD1c, CD1d, ITGAM, ITGAX, ITGB1, CD29, ITG B2 (CD18), ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD 160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Lyl08), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand, cytokine receptor, activating NK cell receptor, or any combination thereof.

[0087] Activation domain As used herein, chimeric antigen receptor can comprise an activation domain.The activation domain can include, but is not limited to, CD3.CD3 is a component of the T cell receptor on natural T cells and has been shown to be an important intracellular activation component in CAR.In a preferred embodiment, CD3 is CD3 zeta.

[0088] Hinge Area As described herein, a chimeric antigen receptor can comprise a hinge region, which is part of the extracellular domain and is sometimes referred to as a "spacer" region. A variety of hinges can be used in accordance with the present invention, including the costimulatory molecules described above, immunoglobulin (Ig) sequences, or other appropriate molecules to achieve a desired specific distance from the target cell. In some embodiments, the entire extracellular domain comprises the hinge region.

[0089] transmembrane region As used herein, a chimeric antigen receptor (CAR) can comprise a transmembrane region / domain. A CAR can be designed to include a transmembrane domain fused to the extracellular domain of the CAR. Similarly, this can be fused to the intracellular domain of the CAR. In one embodiment, a transmembrane domain that is naturally associated with one of the domains in the CAR is used. In some cases, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domain to the transmembrane domain of the same or a different surface membrane protein to minimize interaction with other members of the receptor complex. The transmembrane domain can be derived from either natural or synthetic sources. If the source is natural, the domain can be derived from any membrane-bound or transmembrane protein. Transmembrane regions that are particularly useful in the present invention include CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (alpha, beta, delta, epsilon, gamma, zeta), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, programmed death-1 (PD-1), inducible T-cell costimulator (ICOS), lymphocyte function-associated antigen-1 (LFA-1; CD11a and CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14;TNFSF14), NKG2C, Ig alpha (CD79a), DAP10, Fc gamma receptor, MHC class I molecule, TNFR, integrin, signaling lymphocyte activation molecule, BTLA, Toll ligand receptor, ICAM-1, CDS, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD8 alpha, CD8 beta, IL-2R beta, IL-2R gamma, IL-7R alpha , ITGA4, VLA1, CD49a, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, ITGAE, CD103, ITGAL, CD1a, CD1b, CD1c, CD1d, ITGAM, ITGAX, ITGB1, CD29, ITG B2 (CD18), ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD 160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Lyl08), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand, cytokine receptor, activating NK cell receptor, immunoglobulin protein, or a fragment or any combination thereof;

[0090] immune cells According to certain aspects, the present invention provides a cell that is an immune cell comprising an isolated polynucleotide comprising a nucleotide sequence encoding a CAR provided herein or a vector comprising the isolated polynucleotide. Immune cells comprising the isolated polynucleotide and / or vector of the present invention can be referred to as "engineered immune cells." Preferably, engineered immune cells are derived from a human (are cells of human origin prior to being recombined).

[0091] The engineered immune cells can be, for example, of the lymphoid lineage. Non-limiting examples of lymphoid lineage cells include T cells and natural killer (NK) cells. T cells express the T cell receptor (TCR), with most cells expressing α and β chains, and a smaller population expressing γ and δ chains. T cells useful as engineered immune cells of the present invention include CD4 + or CD8 + T helper cells (CD4 + ), cytotoxic T cells (also called cytotoxic T lymphocytes, CTLs; CD8 + Examples of immune cells include, but are not limited to, immune cells (e.g., immune cells), and memory T cells, including central memory T cells, stem-like memory T cells, and effector memory T cells, natural killer T cells, mucosal-associated invariant T cells, and γδ T cells. Other exemplary immune cells include, but are not limited to, macrophages, antigen-presenting cells (APCs), or any immune cell that expresses an inhibitor of a cell-mediated immune response, such as an immune checkpoint inhibitor pathway receptor (e.g., PD-1). Precursor cells of immune cells that can be used in accordance with the present invention include hematopoietic stem and / or progenitor cells. Hematopoietic stem and / or progenitor cells can be derived from bone marrow, umbilical cord blood, adult peripheral blood after cytokine mobilization, etc., by methods known in the art. The immune cells are engineered to recombinantly express the CAR of the present invention.

[0092] Immune cells and their precursor cells can be isolated by methods known in the art, including commercially available methods (see, e.g., Rowland Jones et al., Lymphocytes: A Practical Approach, Oxford University Press, NY 1999). Sources of immune cells or their precursors include, but are not limited to, peripheral blood, umbilical cord blood, bone marrow, or other sources of hematopoietic cells. Various techniques can be used to separate cells to isolate or enrich for desired immune cells. For example, negative selection methods can be used to remove cells that are not the desired immune cells. Additionally, positive selection methods can be used to isolate or enrich for desired immune cells or their precursors, or positive and negative selection methods can be used in combination. When isolating a specific type of cell, such as a specific T cell, various cell surface markers or combinations of markers (e.g., CD3, CD4, CD8, CD34) can be used to separate the cells.

[0093] Immune cells or their precursor cells can be autologous (autologous) or non-autologous (non-autologous) to the subject to whom they are administered in the treatment methods of the present invention. Autologous cells are isolated from the subject to whom engineered immune cells recombinantly expressing a CAR are administered. Optionally, cells can be obtained by leukapheresis, in which leukocytes are selectively removed from drawn blood, recombined, and then reinfused back into the donor. Alternatively, allogeneic cells from a non-autologous donor can be used. In the case of a non-autologous donor, the cells are sorted and matched with human leukocyte antigens (HLA) to determine the appropriate level of compatibility. For both autologous and non-autologous cells, the cells can be cryopreserved, if necessary, until ready for use.

[0094] Various methods for isolating immune cells that can be used for recombinant expression of the CARs of the invention have been previously described and can be used, including using peripheral blood donor lymphocytes (Sadelain et al., Nat. Rev. Cancer 2003; 3:35-45; Morgan et al., Science 2006; 314:126-9), using lymphocyte cultures derived from tumor-infiltrating lymphocytes (TILs) in tumor biopsies (Panelli et al., J. Immunol. 2000; 164:495-504; Panelli et al., J. Immunol. 2000; 164:4382-92), and using antigen-specific peripheral blood leukocytes selectively expanded in vitro using artificial antigen-presenting cells (AAPCs) or dendritic cells (Dupont et al., Cancer Res. 2005; 65:5417-427; Papanicolaou et al., Blood 2003; 102:2498-505). When using stem cells, the cells can be isolated by methods well known in the art (see, for example, Klug et al., Hematopoietic Stem Cell Protocols, Humana Press, NJ 2002; Freshney et al., Culture of Human Stem Cells, John Wiley & Sons 2007).

[0095] According to certain embodiments, methods of producing engineered immune cells include transfecting or transducing immune effector cells isolated from an individual to express one or more CARs according to embodiments of the present invention. Methods of preparing immune cells for immunotherapy are described, for example, in WO2014 / 130635, WO2013 / 176916, and WO2013 / 176915, which are incorporated herein by reference. Individual steps that can be used in the preparation of engineered immune cells are disclosed, for example, in WO2014 / 039523, WO2014 / 184741, WO2014 / 191128, WO2014 / 184744, and WO2014 / 184143, which are incorporated herein by reference.

[0096] In certain embodiments, immune effector cells, e.g., T cells, are genetically modified with a CAR of the invention (e.g., transformed with a viral vector comprising a nucleic acid encoding the CAR), and then activated and expanded in vitro. In various embodiments, T cells can be activated and expanded before or after genetic modification to express a CAR using, for example, methods described in US 6,352,694, US 6,534,055, US 6,905,680, US 6,692,964, US 5,858,358, US 6,887,466, US 6,905,681, US 7,144,575, US 7,067,318, US 7,172,869, US 7,232,566, US 7,175,843, US 5,883,223, US 6,905,874, US 6,797,514, US 6,867,041, US 2006 / 121005, which are incorporated herein by reference. T cells can be expanded in vitro or in vivo. Generally, T cells of the present invention can be expanded by contact with a surface having attached thereto an agent that stimulates CD3 / TCR complex-associated signals and a ligand that stimulates costimulatory molecules on the surface of the T cells. As a non-limiting example, a population of T cells can be stimulated, as described herein, by contact with, for example, an anti-CD3 antibody, or an antigen-binding fragment thereof, or an anti-CD3 antibody immobilized on a surface, or by contact with a protein kinase C activator (e.g., bryostatin) in combination with a calcium ionophore, or by activation of the CAR itself. To costimulate an accessory molecule on the surface of T cells, a ligand that binds to the accessory molecule is used. For example, a population of T cells can be contacted with an anti-CD3 antibody and an anti-CD28 antibody under conditions appropriate to stimulate T cell proliferation.Suitable conditions for T cell culture include, for example, an appropriate medium (e.g., Minimum Essential Medium or RPMI Media 1640 or X-vivo 5 (Lonza)) that may contain factors necessary for proliferation and survival, including serum (e.g., fetal bovine serum or human serum), cytokines such as IL-2, IL-7, IL-15, and / or IL-21, insulin, IFN-γ, GM-CSF, TGFβ, and / or other additives for cell growth known to those of skill in the art. In other embodiments, T cells can be activated and stimulated with feeder cells and appropriate antibodies and cytokines for expansion using methods such as those described in U.S. Pat. No. 6,040,177, U.S. Pat. No. 5,827,642, and WO2012129514, which are incorporated herein by reference.

[0097] Antibodies and antigen-binding domains The present invention generally relates to isolated anti-TAA antibodies (e.g., anti-TIP-1 antibodies or anti-GPC3 antibodies), chimeric antigen receptors (CARs), nucleic acids and expression vectors encoding the antibodies and CARs, recombinant cells containing the vectors, and compositions comprising the antibodies, CARs, and recombinant cells expressing the CARs. Methods for making the antibodies and CARs, and methods for using the antibodies and CARs to treat diseases, including cancer and inflammatory diseases, are also described. The antigen-binding domains of the antibodies and CARs of the present invention possess one or more desirable functional properties, including, but not limited to, high affinity binding to the TAA, high specificity for the TAA, the ability to stimulate complement-dependent cytotoxicity (CDC), antibody-dependent cellular phagocytosis (ADCP), and / or antibody-dependent cellular cytotoxicity (ADCC) against cells expressing the TAA, and the ability to inhibit tumor growth in subjects and animal models when administered alone or in combination with other anti-cancer therapies.

[0098] In a general aspect, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof that binds to a TAA (eg, TIP-1 or GPC3).

[0099] As used herein, the term "antibody" is used broadly to include immunoglobulins or antibody molecules, including human antibodies, humanized antibodies, composite antibodies, and chimeric antibodies, as well as antibody fragments that are monoclonal or polyclonal. Generally, antibodies are proteins or peptide chains that exhibit binding specificity to a specific antigen. Antibody structure is well known. Immunoglobulins can be assigned to five major classes (i.e., IgA, IgD, IgE, IgG, and IgM) depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4. Thus, antibodies of the present invention can be of any of the five major classes or corresponding subclasses. Preferably, antibodies of the present invention are IgG1, IgG2, IgG3, or IgG4. Antibody light chains of vertebrate species can be assigned to one of two clearly distinct types, kappa and lambda, based on the amino acid sequence of their constant domains. Thus, antibodies of the present invention can contain kappa or lambda light chain constant domains. According to certain embodiments, the antibodies of the present invention comprise heavy and / or light chain constant regions of a rat or human antibody. In addition to the heavy and light chain constant domains, the antibodies contain an antigen-binding region composed of a light chain variable region and a heavy chain variable region, each of which contains three domains (i.e., complementarity-determining regions 1 to 3; CDR1, CDR2, and CDR3). The light chain variable region domains are alternatively referred to as LCDR1, LCDR2, and LCDR3, and the heavy chain variable region domains are alternatively referred to as HCDR1, HCDR2, and HCDR3.

[0100] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to a TAA (e.g., TIP-1 or GPC3) is substantially free of antibodies that do not bind to the same TAA (e.g., TIP-1 or GPC3)). Moreover, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0101] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies of the invention can be produced by hybridoma methods, phage display techniques, single lymphocyte gene cloning techniques, or by recombinant DNA methods. For example, monoclonal antibodies can be produced by hybridomas comprising B cells obtained from a transgenic non-human animal, e.g., a transgenic mouse or rat, whose genome comprises a human heavy chain transgene and a light chain transgene.

[0102] As used herein, the terms "antigen-binding fragment" and / or "antigen-binding domain" refer to antibody fragments such as, for example, diabodies, Fab, Fab', F(ab'), Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized diabodies (ds diabodies), single-chain antibody molecules (scFv), single-domain antibodies (sdab), scFv dimers (bivalent diabodies), multispecific antibodies formed from portions of antibodies comprising one or more CDRs, camelized single-domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment that binds to an antigen but does not comprise the complete antibody structure. An antigen-binding fragment can bind to the same antigen as the parent antibody or parent antibody fragment. According to certain embodiments, the antigen-binding fragment comprises a light chain variable region, a light chain constant region, and the Fd segment of a heavy chain. According to other particular embodiments, the antigen-binding fragment comprises Fab and F(ab'). The antigen-binding domain is capable of binding to the same antigen as the parent antibody. According to particular embodiments, the antigen-binding domain comprises a single-chain antibody molecule (scFv).

[0103] As used herein, the term "single-chain antibody" refers to a conventional single-chain antibody in the art, which comprises a heavy chain variable region and a light chain variable region linked by a short peptide of about 15 to about 20 amino acids. As used herein, the term "single-domain antibody" refers to a conventional single-domain antibody in the art, which comprises a heavy chain variable region and a heavy chain constant region, or comprises only a heavy chain variable region.

[0104] As used herein, the term "human antibody" refers to an antibody produced by a human or an antibody having an amino acid sequence corresponding to an antibody produced by a human, made using any technique known in the art. This definition of a human antibody includes intact or full-length antibodies, fragments thereof, and / or antibodies comprising at least one human heavy and / or light chain polypeptide.

[0105] As used herein, the terms "humanized antibody" and / or "humanized antigen-binding domain" refer to a non-human antibody and / or non-human antigen-binding domain that has been modified to increase sequence homology with a human antibody and / or human antigen-binding domain, such that the antigen-binding properties of the antibody and / or antigen-binding domain are retained but its antigenicity in the human body is reduced.

[0106] As used herein, the terms "chimeric antibody" and / or "chimeric antigen-binding domain" refer to antibodies and / or antigen-binding domains in which the amino acid sequences of the immunoglobulin molecule are derived from two or more species. The variable regions of both the light and heavy chains often correspond to the variable regions of antibodies and / or antigen-binding domains derived from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and capacity, while the constant regions correspond to the sequences of antibodies and / or antigen-binding domains derived from another species of mammal (e.g., human) to avoid eliciting an immune response in that species.

[0107] As used herein, the term "multispecific antibody" refers to an antibody comprising a plurality of immunoglobulin variable domain sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In one embodiment, the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes do not overlap or substantially do not overlap. In one embodiment, the first and second epitopes are on different antigens, e.g., different proteins (or different subunits of a multimeric protein). In one embodiment, the multispecific antibody comprises a third, fourth, or fifth immunoglobulin variable domain. In one embodiment, the multispecific antibody is a bispecific, trispecific, or tetraspecific antibody molecule.

[0108] As used herein, the term "bispecific antibody" refers to a multispecific antibody that binds to no more than two epitopes or two antigens. A bispecific antibody is characterized by a first immunoglobulin variable domain sequence that has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope. In one embodiment, the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes are on different antigens, e.g., different proteins (or different subunits of a multimeric protein). In one embodiment, a bispecific antibody comprises heavy and light chain variable domain sequences that have binding specificity for a first epitope and heavy and light chain variable domain sequences that have binding specificity for a second epitope. In one embodiment, a bispecific antibody comprises a half antibody or fragment thereof having binding specificity for a first epitope and a half antibody or fragment thereof having binding specificity for a second epitope. In one embodiment, a bispecific antibody comprises an scFv or fragment thereof having binding specificity for a first epitope and an scFv or fragment thereof having binding specificity for a second epitope. In one embodiment, the first epitope is located on a TAA of the invention, and the second epitope is located on PD-1, PD-L1, TIM-3, LAG-3, CTLA-4, EGFR, HER-2, CD19, CD20, CD33, CD3, CD73, CD47, TIP-1, GPC3, apelin, DLL3, folate receptor alpha, Claudin18.2, and / or other tumor-associated immunosuppressive factors or surface antigens. In one embodiment, the first and second epitopes are located on the same TAA of the invention.

[0109] As used herein, an antibody and / or antigen-binding domain that "specifically binds to a TAA" has a KD of 1×10 -7 M or less, preferably 1 × 10 -8 M or less, preferably 5×10 -9M or less, 1×10 -9 M or less, 5×10 -10 M or less, or 1 x 10 -10 The term "KD" refers to an antibody and / or antigen-binding domain that binds to a TAA, preferably a human TAA (e.g., TIP-1 or GPC3), with M or less. The term "KD" refers to the dissociation constant, obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and expressed as a molar concentration (M). KD values ​​for antibodies and / or antigen-binding domains can be determined in light of the present disclosure using methods in the art. For example, the KD of an antibody and / or antigen-binding domain can be determined by using surface plasmon resonance, e.g., by using a biosensor system, e.g., a Biacore® system, or by using biolayer interferometry technology, e.g., an Octet RED96 system.

[0110] The smaller the KD value of an antibody and / or antigen-binding domain, the higher the affinity with which the antibody and / or antigen-binding domain binds to a target antigen.

[0111] According to a particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof, or chimeric antigen receptor (CAR) comprising an antigen-binding domain, wherein the monoclonal antibody or antigen-binding fragment thereof, or the antigen-binding domain is (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively; and a heavy chain complementarity-determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity-determining region 1 (LCDR1), LCDR2, and LCDR3, each having a polypeptide sequence as follows:

[0112] According to a particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof, or chimeric antigen receptor (CAR) comprising an antigen-binding domain, wherein the monoclonal antibody or antigen-binding fragment thereof, or the antigen-binding domain is (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55 and 56, respectively; (6) SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; or (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively; and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, each having a polypeptide sequence of:

[0113] According to a particular aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising (1) SEQ ID NOs: 3, 4, 5, 6, 7 and 8, respectively The present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, having a polypeptide sequence of:

[0114] According to a particular aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising (1) SEQ ID NOs: 9, 10, 11, 12, 13 and 14, respectively The present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, having a polypeptide sequence of:

[0115] According to another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof, or chimeric antigen receptor (CAR) comprising an antigen-binding domain, wherein the monoclonal antibody or antigen-binding fragment thereof or antigen-binding domain comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example 95%, 96%, 97%, 98%, or 99% identical to one of SEQ ID NOs: 85, 106, 15, 29, 43, 57, 71, or 1, or a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example 95%, 96%, 97%, 98%, or 99% identical to one of SEQ ID NOs: 86, 107, 16, 30, 44, 58, 72, or 2. According to a preferred embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof or antigen-binding domain thereof of the present invention comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, 71, or 1, or a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, 72, or 2, respectively.

[0116] In another particular aspect, the present invention relates to the isolated monoclonal antibody or antigen-binding fragment thereof or antigen-binding domain thereof of the present invention, a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; g. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; or h. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2 Includes:

[0117] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2.

[0118] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 17, 18, 19, 20, 21, and 22, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 15, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 16. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0119] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 31, 32, 33, 34, 35, and 36, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 29, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 30. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30.

[0120] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 45, 46, 47, 48, 49, and 50, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 43, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 44. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44.

[0121] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 57, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 58. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58.

[0122] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 73, 74, 75, 76, 77, and 78, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 71, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 72. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72.

[0123] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 87, 88, 89, 90, 91, and 92, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 85, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 86. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86.

[0124] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 108, 109, 110, 111, 112, and 113, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 106, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 107. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107.

[0125] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2.

[0126] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 15, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 16. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0127] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 29, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 30. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30.

[0128] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 43, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 44. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44.

[0129] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 57, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 58. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58.

[0130] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 71, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 72. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72.

[0131] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 93, 94, 95, 96, 97, and 98, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 85, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 86. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86.

[0132] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 114, 115, 116, 117, 118, and 119, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 106, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 107. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107.

[0133] In another particular aspect, the present invention relates to the isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, wherein the antibody or antigen-binding fragment thereof is chimeric.

[0134] In another particular aspect, the present invention relates to the isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, wherein the antibody or antigen-binding fragment thereof is human or humanized.

[0135] In another specific embodiment, the humanized monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 99-102, 120-123, 126-132, 139-140, 143-149, 153-156, 160-163, 183-197, or 202-205, or a light chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 103-105, 124-125, 133-138, 141-142, 150-152, 157-159, 164-166, or 198-201.

[0136] In another particular embodiment, the humanized monoclonal antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 Includes:

[0137] In another specific embodiment, the antigen-binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 99-102, 120-123, 126-132, 139-140, 143-149, 153-156, 160-163, 183-197, or 202-205, or a light chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 103-105, 124-125, 133-138, 141-142, 150-152, 157-159, 164-166, or 198-201.

[0138] According to another particular embodiment, the antigen-binding domain is humanized, (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 Includes:

[0139] In another particular embodiment, the antigen-binding domain is a single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3.

[0140] In certain embodiments, the encoded antigen-binding domain is a humanized single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3. In certain embodiments, the antigen-binding domain is a humanized single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3. In certain embodiments, the humanized single-chain variable fragment (scFv) comprises a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 167-182. In certain embodiments, the humanized single-chain variable fragment (scFv) comprises a polypeptide sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 167-182.

[0141] According to another particular embodiment, the chimeric antigen receptor comprises one or more antigen-binding domains.

[0142] According to another particular embodiment, the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0143] In another general aspect, the present invention relates to an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof of the present invention. In another general aspect, the present invention relates to an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR comprises the antigen-binding domain of the present invention. Those skilled in the art will understand that the coding sequence of a protein can be altered (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Thus, those skilled in the art will understand that the nucleic acid sequence encoding the monoclonal antibody or antigen-binding fragment thereof of the present invention can be modified without changing the amino acid sequence of the protein.

[0144] In another general aspect, the present invention relates to a vector comprising an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, and / or a CAR of the present invention. In light of the present disclosure, any vector known to those of skill in the art can be used, for example, a plasmid, cosmid, phage vector, or viral vector. In some embodiments, the vector is a recombinant expression vector, for example, a plasmid. The vector can include any elements for establishing the conventional functions of an expression vector, such as a promoter, a ribosome-binding element, a terminator, an enhancer, a selection marker, and an origin of replication. The promoter can be a constitutive, inducible, or repressible promoter. Numerous expression vectors capable of delivering nucleic acids into cells are known in the art and can be used herein to produce antibodies or antigen-binding fragments thereof in cells. Conventional cloning techniques or artificial gene synthesis can be used to generate recombinant expression vectors according to embodiments of the present invention.

[0145] In another general aspect, the present invention relates to a host cell comprising an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof of the present invention. Any host cell known to one of skill in the art in light of the present disclosure can be used for recombinant expression of an antibody or antigen-binding fragment thereof of the present invention. In some embodiments, the host cell is an Escherichia coli (E. coli) TG1 or BL21 cell (e.g., for expression of scFv or Fab antibodies), a CHO-DG44 or CHO-K1 cell, or a HEK293 cell (e.g., for expression of full-length IgG antibodies). In certain embodiments, the recombinant expression vector is transformed into the host cell by conventional methods, e.g., chemical transfection, heat shock, or electroporation, where it is stably integrated into the host cell genome for efficient expression of the recombinant nucleic acid.

[0146] In another general aspect, the invention relates to a method for producing a monoclonal antibody or antigen-binding fragment thereof and / or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising culturing cells containing nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof or the bispecific antibody or antigen-binding fragment thereof under conditions to produce the monoclonal antibody or antigen-binding fragment thereof or the bispecific antibody or antigen-binding fragment thereof of the invention, and recovering the antibody or antigen-binding fragment thereof from the cells or cell culture (e.g., from the supernatant). The expressed antibody or antigen-binding fragment thereof can be recovered from the cells and purified as described herein according to routine techniques known in the art.

[0147] In another general aspect, the invention relates to a cell transduced with a vector comprising an isolated nucleic acid encoding a CAR of the invention. The terms "transduced" or "transduction" refer to the process by which exogenous nucleic acid is transferred or introduced into a host cell. A "transduced" cell is a cell that has been transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny. In certain embodiments, the cell is a CAR-T cell, preferably a human CAR-T cell, where the T cell is engineered to express a CAR of the invention to treat a disease, e.g., cancer. In certain embodiments, the cell is a CAR-NK cell, preferably a human CAR-NK cell, where the NK cell engineered to express a CAR of the invention is used to treat a disease, e.g., cancer.

[0148] In another general aspect, the invention features a method of generating a CAR-T cell by transducing a T cell with a vector comprising an isolated nucleic acid encoding a CAR of the invention.

[0149] In another general aspect, the invention relates to a method of producing a chimeric antigen receptor (CAR) T cell of the invention, comprising culturing T cells comprising a nucleic acid encoding the CAR under CAR-T cell-producing conditions, and recovering the CAR-T cell.

[0150] In another general aspect, the invention features a method of generating CAR-NK cells by transducing NK cells with a vector comprising an isolated nucleic acid encoding a CAR of the invention.

[0151] In another general aspect, the invention features a method of producing a chimeric antigen receptor (CAR)-NK cell of the invention, comprising culturing NK cells comprising a nucleic acid encoding the CAR under CAR-NK cell-producing conditions, and recovering the CAR-NK cell.

[0152] In another general aspect, the invention relates to a method of generating a population of RNA engineered cells comprising a chimeric antigen receptor (CAR) of the invention. The method comprises contacting the cell population with an isolated polynucleotide comprising a nucleic acid encoding a CAR of the invention, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

[0153] Pharmaceutical Composition In another general aspect, the invention relates to a pharmaceutical composition comprising an isolated monoclonal antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, an isolated polynucleotide, an isolated polypeptide, a host cell, and / or an engineered immune cell of the invention, and a pharmaceutically acceptable carrier.

[0154] As used herein, the term "pharmaceutical composition" refers to a product comprising an isolated polynucleotide of the invention, an isolated polypeptide of the invention, a host cell of the invention, an engineered immune cell of the invention, an anti-TIP-1 or anti-GPC3 monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody of the invention, together with a pharmaceutically acceptable carrier. The polynucleotides, polypeptides, host cells, engineered immune cells, anti-TIP-1 or anti-GPC3 monoclonal antibodies or antigen-binding fragments thereof, and / or bispecific antibodies of the invention and compositions comprising them are also useful in the manufacture of medicaments for the therapeutic applications described herein.

[0155] As used herein, the term "carrier" refers to any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, oil, lipid, lipid-containing vesicle, microsphere, liposome encapsulation, or other material known in the art for use in pharmaceutical formulations. It is understood that the characteristics of the carrier, excipient, or diluent will depend on the route of administration for a particular application. As used herein, the term "pharmaceutically acceptable carrier" refers to a non-toxic material that does not interfere with the efficacy or biological activity of the compositions according to the present invention. In certain embodiments, any pharmaceutically acceptable carrier suitable for use in antibody pharmaceutical compositions, in light of the present disclosure, can be used in the present invention.

[0156] Formulation of pharmaceutically active ingredients with pharmaceutically acceptable carriers is known in the art, e.g., Remington: The Science and Practice of Pharmacy (e.g., 21st Edition (2005) and any later editions). Non-limiting examples of additional ingredients include: buffers, diluents, solvents, tonicity adjusters, preservatives, stabilizers, and chelating agents. One or more pharmaceutically acceptable carriers can be used in formulating the pharmaceutical compositions of the present invention.

[0157] In one embodiment of the present invention, the pharmaceutical composition is a liquid formulation. A preferred example of a liquid formulation is an aqueous formulation, i.e., a formulation containing water. Liquid formulations can include solutions, suspensions, emulsions, microemulsions, gels, etc. Aqueous formulations typically contain at least 50% w / w water, or at least 60%, 70%, 75%, 80%, 85%, 90%, or at least 95% w / w water.

[0158] In one embodiment, the pharmaceutical composition can be formulated as an injectable that can be injected, for example, via an injection device (e.g., a syringe or an infusion pump). The injection can be delivered, for example, subcutaneously, intramuscularly, intraperitoneally, intravitreally, or intravenously.

[0159] In another embodiment, the pharmaceutical composition is a solid formulation, such as a freeze-dried or spray-dried composition that can be used as is, or a solid formulation to which the physician or patient can add solvents and / or diluents before use. Solid dosage forms can include tablets, for example, compressed tablets, and / or coated tablets, and capsules (e.g., hard or soft gelatin capsules). Pharmaceutical compositions can also be in the form of, for example, sachets, dragees, powders, granules, lozenges, or powders for reconstitution.

[0160] Dosage forms may be immediate release, in which case they may include a water soluble or dispersible carrier, or they may be delayed, sustained, or modified release, in which case they may include a water insoluble polymer that modulates the dissolution rate of the dosage form in the gastrointestinal tract or under the skin.

[0161] In other embodiments, the pharmaceutical composition may be delivered intranasally, bucally, or sublingually.

[0162] The pH of the aqueous formulation can be between pH 3 and pH 10. In one embodiment of the invention, the pH of the formulation is from about 7.0 to about 9.5. In another embodiment of the invention, the pH of the formulation is from about 3.0 to about 7.0.

[0163] In another embodiment of the present invention, the pharmaceutical composition comprises a buffering agent. Non-limiting examples of buffering agents include arginine, aspartic acid, bicine, citrate, disodium hydrogen phosphate, fumaric acid, glycine, glycylglycine, histidine, lysine, maleic acid, malic acid, sodium acetate, sodium carbonate, sodium dihydrogen phosphate, sodium phosphate, succinic acid, tartaric acid, tricine, and tris(hydroxymethyl)aminomethane, and mixtures thereof. The buffering agent can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific buffering agents constitute alternative embodiments of the present invention.

[0164] In another embodiment of the present invention, the pharmaceutical composition comprises a preservative. Non-limiting examples of preservatives include benzethonium chloride, benzoic acid, benzyl alcohol, bronopol, butyl 4-hydroxybenzoate, chlorobutanol, chlorocresol, chlorhexidine, chlorphenesin, o-cresol, m-cresol, p-cresol, ethyl 4-hydroxybenzoate, imidurea, methyl 4-hydroxybenzoate, phenol, 2-phenoxyethanol, 2-phenylethanol, propyl 4-hydroxybenzoate, sodium dehydroacetate, thiomerosal, and mixtures thereof. The preservatives can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific preservatives constitute alternative embodiments of the present invention.

[0165] In another embodiment of the present invention, the pharmaceutical composition comprises an isotonicity agent. Non-limiting examples of isotonicity agents include salts (e.g., sodium chloride), amino acids (e.g., glycine, histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, and threonine), alditols (e.g., glycerol, 1,2-propanediol, propylene glycol), 1,3-propanediol, and 1,3-butanediol), polyethylene glycols (e.g., PEG 400), and mixtures thereof. Another example of an isotonicity agent includes a sugar. Non-limiting examples of sugars may be monosaccharides, disaccharides, or polysaccharides, or water-soluble glucans, such as fructose, glucose, mannose, sorbose, xylose, maltose, lactose, sucrose, trehalose, dextran, pullulan, dextrin, cyclodextrin, alpha- and beta-HPCD, soluble starch, hydroxyethyl starch, and sodium carboxymethylcellulose. Another example of an isotonicity agent is a sugar alcohol, where the term "sugar alcohol" is defined as a C(4-8) hydrocarbon having at least one -OH group. Non-limiting examples of sugar alcohols include mannitol, sorbitol, inositol, galactitol, dulcitol, xylitol, and arabitol. The isotonicity agent can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific isotonicity agents constitute alternative embodiments of the invention.

[0166] In another embodiment of the invention, the pharmaceutical composition comprises a chelating agent. Non-limiting examples of chelating agents include citric acid, aspartic acid, salts of ethylenediaminetetraacetic acid (EDTA), and mixtures thereof. The chelating agent can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific chelating agents constitute alternative embodiments of the invention.

[0167] In another embodiment of the invention, the pharmaceutical composition comprises a stabilizer, non-limiting examples of which include one or more aggregation inhibitors, one or more oxidation inhibitors, one or more surfactants, and / or one or more protease inhibitors.

[0168] In another embodiment of the present invention, the pharmaceutical composition comprises a stabilizer, which is carboxy- / hydroxycellulose and its derivatives (e.g., HPC, HPC-SL, HPC-L, and HPMC), cyclodextrin, 2-methylthioethanol, polyethylene glycol (e.g., PEG3350), polyvinyl alcohol (PVA), polyvinylpyrrolidone, salts (e.g., sodium chloride), sulfur-containing substances (e.g., monothioglycerol), or thioglycolic acid. The stabilizer can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific stabilizers constitute alternative embodiments of the present invention.

[0169] In a further embodiment of the present invention, the pharmaceutical composition comprises one or more surfactants, preferably one surfactant, at least one surfactant, or two different surfactants. The term "surfactant" refers to any molecule or ion composed of a water-soluble (hydrophilic) portion and a fat-soluble (lipophilic) portion. The surfactant may be selected, for example, from the group consisting of anionic surfactants, cationic surfactants, nonionic surfactants, and / or zwitterionic surfactants. The surfactant may be present individually or in aggregates at a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific surfactants constitute alternative embodiments of the present invention.

[0170] In a further embodiment of the invention, the pharmaceutical composition comprises one or more protease inhibitors, such as EDTA and / or benzamidine hydrochloride (HCl). The protease inhibitors can be present individually or in aggregates at a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific protease inhibitors constitute alternative embodiments of the invention.

[0171] In another general aspect, the invention relates to a method for producing a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising combining the monoclonal antibody or antigen-binding fragment thereof, and / or the bispecific antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

[0172] How to use In another general aspect, the invention relates to a method of treating cancer in a subject in need thereof, comprising administering to the subject CAR-T cells and / or CAR-NK cells of the invention. The cancer can be selected from, for example, but not limited to, lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors.

[0173] In another general aspect, the present invention relates to a method of targeting a TAA (e.g., TIP-1 or GPC3) on the surface of cancer cells in a subject to achieve cell killing, the method comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof, that specifically binds to the TAA, or a pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof, and / or the bispecific antibody or antigen-binding fragment thereof of the present invention. Binding of the TAA monoclonal or bispecific antibody or antigen-binding fragment to the TAA can mediate complement-dependent cytotoxicity (CDC), antibody-dependent cellular phagocytosis (ADCP), and / or antibody-dependent cellular cytotoxicity (ADCC), or other effects that result in the death of the targeted cancer cells. The monoclonal or bispecific antibody or antigen-binding fragment thereof can, for example, serve to recruit a conjugated drug and / or can form a bispecific antibody with another monoclonal antibody to mediate the death of the targeted cancer cells.

[0174] The functional activity of antibodies and antigen-binding fragments thereof that bind to a TAA (e.g., TIP-1 or GPC3) can be characterized by methods known in the art and by methods described herein. Methods for characterizing antibodies and antigen-binding fragments thereof that bind to a TAA include, but are not limited to, affinity and specificity assays, including Biacore, ELISA, and OctetRed analysis, and detecting binding of antibodies and antigen-binding fragments to the TAA on cells (either cells transfected with the TAA or cells that naturally express the TAA) by FACS. According to certain embodiments, methods for characterizing antibodies and antigen-binding fragments thereof that bind to a TAA include those described below.

[0175] In another general aspect, the present invention relates to a method of treating cancer in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof that specifically binds to a TAA (e.g., TIP-1 or GPC3), or a pharmaceutical composition of the invention. The cancer can be selected from, for example, but not limited to, lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors.

[0176] In another general aspect, the present invention relates to a method of treating an inflammatory disease in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof that specifically binds to a TAA (e.g., TIP-1 or GPC3), or a pharmaceutical composition of the invention.

[0177] According to an embodiment of the present invention, the CAR-T cells or CAR-NK cells comprise a therapeutically effective amount of the expressed CAR of the present invention, and the pharmaceutical composition comprises a therapeutically effective amount of an anti-TAA antibody or an antigen-binding fragment thereof (e.g., an anti-TIP-1 antibody or an anti-GPC3 antibody). As used herein, the term "therapeutically effective amount" refers to the amount of an active ingredient or component that elicits a desired biological or pharmaceutical response in a subject. The therapeutically effective amount can be determined empirically and routinely for the specified purpose.

[0178] As used herein with respect to a CAR, a therapeutically effective amount refers to the amount of a CAR molecule expressed in a transduced T cell or NK cell that modulates an immune response in a subject in need thereof. Further, as used herein with respect to a CAR, a therapeutically effective amount refers to the amount of a CAR molecule expressed in a transduced T cell or NK cell that provides treatment for a disease, disorder, or condition, prevents or slows the progression of the disease, disorder, or condition, or reduces or completely alleviates symptoms associated with the disease, disorder, or condition.

[0179] As used herein with respect to CAR-T cells or CAR-NK cells, a therapeutically effective amount refers to the amount of CAR-T cells or CAR-NK cells that modulates an immune response in a subject in need thereof. Further, as used herein with respect to CAR-T cells or CAR-NK cells, a therapeutically effective amount refers to the amount of CAR-T cells or CAR-NK cells that provides treatment for a disease, disorder, or condition, prevents or slows the progression of the disease, disorder, or condition, or reduces or completely alleviates symptoms associated with the disease, disorder, or condition.

[0180] As used herein with respect to an anti-TAA antibody or antigen-binding fragment thereof, a therapeutically effective amount refers to the amount of an anti-TAA antibody or antigen-binding fragment thereof that modulates an immune response in a subject in need thereof. Also, as used herein with respect to an anti-TAA antibody or antigen-binding fragment thereof, a therapeutically effective amount refers to the amount of an anti-TAA antibody or antigen-binding fragment thereof that provides treatment for a disease, disorder, or condition, prevents or slows the progression of the disease, disorder, or condition, or reduces or completely alleviates symptoms associated with the disease, disorder, or condition.

[0181] According to certain embodiments, the disease, disorder or condition to be treated is cancer, preferably a cancer selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, bile duct cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors. According to other certain embodiments, the disease, disorder or condition to be treated is an inflammatory disease.

[0182] According to certain embodiments, a therapeutically effective amount refers to the amount of therapy that is sufficient to achieve one, two, three, four, or more of the following effects: (i) reducing or ameliorating the severity of the disease, disorder, or condition being treated or the symptoms associated therewith; (ii) reducing the duration of the disease, disorder, or condition being treated or the symptoms associated therewith; (iii) preventing the progression of the disease, disorder, or condition being treated or the symptoms associated therewith; (iv) causing regression of the disease, disorder, or condition being treated or the symptoms associated therewith; (v) preventing the development or onset of the disease, disorder, or condition being treated or the symptoms associated therewith. (vi) preventing the recurrence of the disease, disorder, or condition to be treated or symptoms associated therewith; (vii) reducing hospitalization of a subject having the disease, disorder, or condition to be treated or symptoms associated therewith; (viii) reducing the length of hospitalization of a subject having the disease, disorder, or condition to be treated or symptoms associated therewith; (ix) increasing survival of a subject having the disease, disorder, or condition to be treated or symptoms associated therewith; (xi) suppressing or reducing the disease, disorder, or condition to be treated or symptoms associated therewith in a subject; and / or (xii) enhancing or improving the prophylactic or therapeutic efficacy of another therapy.

[0183] The therapeutically effective amount or dosage can vary depending on a variety of factors, such as the disease, disorder, or condition to be treated, the means of administration, the target site, the physiological state of the subject (including, for example, age, weight, and health), whether the subject is human or animal, other pharmaceutical agents administered, and whether the treatment is prophylactic or therapeutic. Treatment dosages are optimally titrated to optimize safety and efficacy.

[0184] According to certain embodiments, the compositions described herein are formulated to be suitable for the intended route of administration to a subject. For example, the compositions described herein can be formulated to be suitable for intravenous, subcutaneous, or intramuscular administration.

[0185] The cells of the present invention can be administered by any convenient method known to those skilled in the art. For example, the cells of the present invention can be administered to a subject by aerosol inhalation, injection, ingestion, transfusion, implantation, and / or transplantation. Compositions comprising the cells of the present invention can be administered intraarterially, subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, intrapleurally, by intravenous (iv) injection, or intraperitoneally. In certain embodiments, the cells of the present invention can be administered with or without lymphodepletion of the subject.

[0186] Pharmaceutical compositions comprising the cells of the invention expressing a CAR of the invention can be provided in sterile liquid formulations, generally isotonic aqueous solutions with cell suspensions, or optionally as emulsions, dispersions, etc., which are generally buffered to a selected pH. The compositions can include carriers suitable for cell integrity and viability and for administration of the cell composition, such as water, saline, phosphate-buffered saline, etc.

[0187] Sterile injectable solutions can be prepared by incorporating the cells of the present invention in a suitable solvent in a suitable amount, along with various other ingredients, as desired. Such compositions can contain pharmaceutically acceptable carriers, diluents, or excipients, such as sterile water, physiological saline, glucose, dextrose, etc., that are suitable for use in cell compositions and for administration to subjects, e.g., humans. Buffers suitable for providing cell compositions are well known in the art. Any vehicle, diluent, or additive used will be compatible with preserving the integrity and viability of the cells of the present invention.

[0188] The cells of the present invention can be administered in any physiologically acceptable vehicle. Cell populations containing the cells of the present invention can include purified cell populations. Those skilled in the art can easily determine the purity of cells in a cell population using various well-known methods. The purity range of a cell population containing genetically modified cells of the present invention can be about 50% to about 55%, about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, or about 95% to about 100%. Dosages can be easily adjusted by those skilled in the art; for example, a decrease in purity may require an increase in dosage.

[0189] The cells of the invention are generally administered as a dose based on the number of cells per kilogram of body weight (cells / kg) of the subject to whom the cells are administered. Generally, cell doses are about 10 per kg of body weight, depending on the mode and location of administration. 4 ~about 10 10 Cell numbers, e.g., about 10 5 ~about 10 9 , about 10 5 ~about 10 8 , about 10 5 ~about 10 7 or about 10 5 ~about 10 6 Generally, for systemic administration, higher doses are used than for local administration, where the immune cells of the invention are administered within the region of the tumor and / or cancer. Exemplary dose ranges include, but are not limited to, 1 x 10 4~1×10 8 、2×10 4 ~1×10 8 、3×10 4 ~1×10 8 、4×10 4 ~1×10 8 、5×10 4 ~6×10 8 、7×10 4 ~1×10 8 、8×10 4 ~1×10 8 、9×10 4 ~1×10 8 、1×10 5 ~1×10 8 、1×10 5 ~9×10 7 、1×10 5 ~8×10 7 、1×10 5 ~7×10 7 、1×10 5 ~6×10 7 、1×10 5 ~5×10 7 、1×10 5 ~4×10 7 、1×10 5 ~4×10 7 、1×10 5 ~3×10 7 、1×10 5 ~2×10 7 、1×10 5 ~1×10 7 、1×10 5 ~9×10 6 、1×10 5 ~8×10 6 、1×10 5 ~7×10 6 、1×10 5 ~6×10 6 、1×10 5 ~5×10 6 、1×10 5 ~4×10 6 、1×10 5 ~4×10 6 、1×10 5 ~3×10 6 、1×10 5 ~2×10 6 、1×105 ~1×10 6 , 2 × 10 5 ~9×10 7 , 2 × 10 5 ~8×10 7 , 2 × 10 5 ~7×10 7 , 2 × 10 5 ~6×10 7 , 2 × 10 5 ~5×10 7 , 2 × 10 5 ~4×10 7 , 2 × 10 5 ~4×10 7 , 2 × 10 5 ~3×10 7 , 2 × 10 5 ~2×10 7 , 2 × 10 5 ~1×10 7 , 2 × 10 5 ~9×10 6 , 2 × 10 5 ~8×10 6 , 2 × 10 5 ~7×10 6 , 2 × 10 5 ~6×10 6 , 2 × 10 5 ~5×10 6 , 2 × 10 5 ~4×10 6 , 2 × 10 5 ~4×10 6 , 2 × 10 5 ~3×10 6 , 2 × 10 5 ~2×10 6 , 2 × 10 5 ~1×10 6 , 3×10 5 ~3×10 6 These include the number of cells / kg, etc. Additionally, the dose can be adjusted to account for whether a single dose or multiple doses are administered. The precise determination of what is considered an effective dose can be based on individual factors for each subject.

[0190] As used herein, the terms "treat," "treating," and "treatment" all refer to an improvement or reversal of at least one measurable physical parameter associated with a cancer and / or inflammatory disease, disorder, or condition, which may, but is not necessarily, discernible in the subject. The terms "treat," "treating," and "treatment" can also refer to causing regression, preventing progression, or at least slowing progression of a disease, disorder, or condition. In a particular embodiment, "treat," "treating," and "treatment" refer to alleviating, preventing the development or onset, or reducing the duration of one or more symptoms associated with a disease, disorder, or condition, such as a tumor, more preferably a cancer. In a particular embodiment, "treat," "treating," and "treatment" refer to preventing the recurrence of a disease, disorder, or condition. In a particular embodiment, "treat," "treating," and "treatment" refer to increasing the survival of a subject having a disease, disorder, or condition. In a particular embodiment, "treat," "treating," and "treatment" refer to eliminating a disease, disorder, or condition in a subject.

[0191] Certain embodiments provide compositions for use in the treatment of cancer and / or inflammatory diseases, disorders, or conditions. For cancer therapy, the provided compositions can be used in combination with another treatment, including, but not limited to, chemotherapy, anti-CD20 mAb, anti-TIM-3 mAb, anti-LAG-3 mAb, anti-EGFR mAb, anti-HER-2 mAb, anti-CD19 mAb, anti-CD33 mAb, anti-CD47 mAb, anti-CD73 mAb, anti-DLL-3 mAb, anti-Apelin mAb, anti-FOLR1 mAb, anti-CTLA-4 mAb, anti-PD-L1 mAb, anti-PD-1 mAb, anti-Claudin18.2 mAb, other immuno-oncology agents, anti-angiogenic agents, radiation therapy, antibody-drug conjugates (ADCs), targeted therapies, or other anti-cancer agents. An antibody against a given TAA can be used to construct a bispecific antibody with a partner mAb against PD-1, PD-L1, LAG3, TIM-3, CTLA-4, EGFR, HER-2, CD19, CD20, CD33, CD73, CD47, CD3, Apelin, DLL-3, TIP-1, GPC3, Claudin18.2, folate receptor alpha (FOLR1), and / or a second TAA to treat cancers / tumors expressing both TAAs. Two antibodies that recognize two different epitopes on the same TAA can also be used to construct a bispecific antibody to treat cancers / tumors expressing TAAs.

[0192] According to certain embodiments, a method of treating cancer in a subject in need thereof comprises administering to the subject the CAR-T cells and / or CAR-NK cells of the invention in combination with an agent that increases the efficacy of cells expressing a CAR molecule, including, but not limited to, an antibody fragment that binds to CD73, CD39, PD1, PD-L1, PD-L2, CTLA4, TIM3, or LAG3, or an adenosine A2a receptor antagonist.

[0193] According to certain embodiments, a method of treating cancer in a subject in need thereof comprises administering to the subject the CAR-T cells and / or CAR-NK cells of the invention in combination with an agent that ameliorates one or more side effects associated with the administration of cells expressing a CAR molecule. Such agents include, but are not limited to, steroids, inhibitors of TNFα, or inhibitors of IL-6.

[0194] According to certain embodiments, a method of treating cancer in a subject in need thereof comprises administering to the subject the CAR-T cells and / or CAR-NK cells of the invention in combination with an agent that treats a disease associated with GPC3, including, but not limited to, an anti-GPC3 monoclonal antibody or bispecific antibody.

[0195] As used herein, in the context of administering two or more therapies to a subject, the term "in combination" refers to the use of multiple therapies.The use of the term "in combination" does not limit the order in which therapies are administered to a subject.For example, a first therapy (e.g., a composition described herein) can be administered before (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks before), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks after) the administration of a second therapy to a subject.

[0196] In another general aspect, the invention relates to a method for determining the level of a TAA (e.g., TIP-1 or GPC3) in a subject, the method comprising: (a) obtaining a sample from the subject; (b) contacting the sample with a monoclonal antibody or antigen-binding fragment thereof of the invention; and (c) determining the level of the TAA in the subject.

[0197] As used herein, "sample" refers to a biological sample isolated from a subject, and can include, but is not limited to, whole blood, serum, plasma, blood cells, endothelial cells, tissue biopsy (e.g., cancer tissue), lymph, ascites, interstitial fluid, bone marrow, cerebrospinal fluid, saliva, mucus, sputum, sweat, urine, or any other secretion, excretion, or other bodily fluid. A "blood sample" refers to whole blood or any fraction thereof, including blood cells, serum, and plasma.

[0198] In certain embodiments, the level of a TAA (e.g., TIP-1 or GPC3) in a subject can be determined using an assay selected from, but not limited to, Western blot assay, immunohistochemistry (IHC), and ELISA assay. Relative protein levels can be determined using Western blot analysis and IHC, and absolute protein levels can be determined using ELISA assay. When determining the relative level of a TAA, the level of the TAA can be determined between at least two samples, for example, between samples from the same subject at different time points, between samples from different tissues within the same subject, and / or between samples from different subjects. Alternatively, when determining the absolute level of a TAA, for example, by ELISA assay, the absolute level of the TAA in a sample can be determined by preparing a standard for the ELISA assay before testing the sample. Those skilled in the art will understand the analytical techniques to be used to determine the level of a TAA in a sample from a subject using the antibody or antigen-binding fragment thereof of the present invention.

[0199] Utilizing a method for determining the level of a TAA (e.g., TIP-1 or GPC3) in a sample from a subject can result in the diagnosis of abnormal (elevated, decreased, or insufficient) TAA levels in a disease, allowing appropriate treatment decisions to be made. Such diseases can be selected from, but are not limited to, cancer and inflammatory diseases. Furthermore, by monitoring the level of a TAA in a subject, the risk of developing such a disease can be determined based on knowledge of the level of the TAA in a particular disease and / or during the progression of a particular disease.

[0200] Embodiment The present invention also provides the following non-limiting embodiments.

[0201] Embodiment 1 is (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively; an isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence shown below; wherein the antibody or antigen-binding fragment thereof specifically binds to GPC3, preferably human GPC3.

[0202] In embodiment 2, (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55 and 56, respectively; (6) SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; or (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively; an isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence shown below; wherein the antibody or antigen-binding fragment thereof specifically binds to GPC3, preferably human GPC3.

[0203] Embodiment 3 is (1) SEQ ID NOs: 3, 4, 5, 6, 7 and 8, respectively An isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0204] Embodiment 4 is (1) SEQ ID NOs: 9, 10, 11, 12, 13 and 14, respectively An isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0205] Embodiment 5 is an isolated monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 4, comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, 71, or 1, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, 72, or 2.

[0206] Embodiment 6 is (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; or (8) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2 The isolated monoclonal antibody or antigen-binding fragment of any one of embodiments 1 to 5, comprising:

[0207] Embodiment 7 is the isolated monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 6, wherein the antibody or antigen-binding fragment thereof is chimeric and / or human or humanized.

[0208] Embodiment 8 is a humanized monoclonal antibody or antigen-binding fragment thereof, comprising a heavy chain variable region having a polypeptide sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 99-102, 120-123, 126-132, 139-140, 143-149, 153-156, 160-163, 183-197, or 202-205; Alternatively, it is an isolated monoclonal antibody or antigen-binding fragment of embodiment 7, comprising a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 103-105, 124-125, 133-138, 141-142, 150-152, 157-159, 164-166, or 198-201.

[0209] Embodiment 9 is a method for producing a humanized monoclonal antibody or antigen-binding fragment thereof, comprising: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 The isolated monoclonal antibody or antigen-binding fragment thereof of embodiment 8, comprising:

[0210] Embodiment 10 is the isolated monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 9, wherein the isolated antibody or antigen-binding fragment thereof is capable of inducing effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or mediating the recruitment of a conjugated agent, and / or forming a bispecific antibody with another mAb or antigen-binding fragment thereof that has cancericidal activity.

[0211] Embodiment 11 is an isolated bispecific antibody or antigen-binding fragment thereof comprising the monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 10.

[0212] Embodiment 12 is an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment of any one of Embodiments 1-10.

[0213] Embodiment 13 is an isolated nucleic acid encoding the bispecific antibody or antigen-binding fragment thereof of Embodiment 11.

[0214] Embodiment 14 is a vector comprising the isolated nucleic acid of embodiment 12 or 13.

[0215] Embodiment 15 is a host cell comprising the vector of embodiment 14.

[0216] Embodiment 16 is a pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 10, or the bispecific antibody or antigen-binding fragment thereof of Embodiment 11, and a pharmaceutically acceptable carrier.

[0217] Embodiment 17 is a method of targeting a TAA on the surface of cancer cells and / or treating cancer and / or treating an inflammatory disease in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of Embodiment 16, optionally wherein the cancer is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors.

[0218] Embodiment 18 is a method for producing the monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 10, or the bispecific antibody or antigen-binding fragment thereof of Embodiment 11, the method comprising culturing cells containing a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, under conditions for producing the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, and recovering the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof from the cells or culture.

[0219] Embodiment 19 is a method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 10, or the bispecific antibody or antigen-binding fragment thereof of Embodiment 11, comprising combining the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

[0220] Embodiment 20 is a method for determining the level of a TAA in a subject, comprising: a. obtaining a sample from a subject; b. contacting the sample with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-10; and c. Determining the level of TAA in a subject The method includes:

[0221] Embodiment 21 is the method of embodiment 20, wherein the sample is a tissue sample or a blood sample, and optionally the tissue sample is a cancer tissue sample.

[0222] Embodiment 22 is an isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an extracellular domain comprising at least one antigen-binding domain that specifically binds to GPC3; (b) hinge region; (c) a transmembrane region, and (d) Intracellular signaling domain It is an isolated polynucleotide comprising:

[0223] Embodiment 23 is a method for producing an antigen-binding domain comprising: (1) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, respectively; (2) SEQ ID NOs: 108, 109, 110, 111, 112 and 113, respectively; (3) SEQ ID NOs: 17, 18, 19, 20, 21 and 22, respectively; (4) SEQ ID NOs: 31, 32, 33, 34, 35 and 36, respectively; (5) SEQ ID NOs: 45, 46, 47, 48, 49 and 50, respectively; (6) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively; or (7) SEQ ID NOs: 73, 74, 75, 76, 77 and 78, respectively; 23. The isolated polynucleotide of embodiment 22, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0224] Embodiment 24 is a method for producing an antigen-binding domain comprising: (1) SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (2) SEQ ID NOs: 114, 115, 116, 117, 118 and 119, respectively; (3) SEQ ID NOs: 23, 24, 25, 26, 27 and 28, respectively; (4) SEQ ID NOs: 37, 38, 39, 40, 41 and 42, respectively; (5) SEQ ID NOs: 51, 52, 53, 54, 55 and 56, respectively; (6) SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; or (7) SEQ ID NOs: 79, 80, 81, 82, 83 and 84, respectively; 23. The isolated polynucleotide of embodiment 22, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0225] Embodiment 25 is the isolated polynucleotide of any one of Embodiments 22 to 24, wherein the antigen-binding domain comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 85, 106, 15, 29, 43, 57, or 71, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 86, 107, 16, 30, 44, 58, or 72.

[0226] Embodiment 26 is a method for producing an antigen-binding domain comprising: a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 106 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 107; c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; or g. A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72 26. The isolated polynucleotide of embodiment 25, comprising:

[0227] Embodiment 27 is the isolated polynucleotide of any one of Embodiments 22-24, wherein the antigen-binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 99-102, 120-123, 126-132, 139-140, 143-149, 153-156, 160-163, 183-197, or 202-205, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 103-105, 124-125, 133-138, 141-142, 150-152, 157-159, 164-166, or 198-201.

[0228] Embodiment 28 is an antibody wherein the antigen-binding domain is humanized; (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 121 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 122 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 125; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 123 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 124; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 139 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 140 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 153 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 157; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 154 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 158; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 147 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 149 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 152; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 160 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 161 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 163 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 205 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 202 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 164; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (58) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (59) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (60) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (61) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (62) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (63) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (64) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (65) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (66) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (67) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (68) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (69) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (70) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (71) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (72) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (73) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (74) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; or (75) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 28. The isolated polynucleotide of embodiment 27, comprising:

[0229] Embodiment 29 is the isolated polynucleotide of any one of Embodiments 22 to 28, wherein the antigen-binding domain is a single-chain variable fragment (scFv) that specifically binds to GPC3, preferably human GPC3.

[0230] Embodiment 30 is the isolated polynucleotide of embodiment 29, wherein the single chain variable fragment (scFv) is humanized.

[0231] Embodiment 31 is the isolated polynucleotide of embodiment 29 or 30, wherein the single chain variable fragment (scFv) comprises a polypeptide sequence at least 95% identical to any one of SEQ ID NOs: 167-182.

[0232] Embodiment 32 is the isolated polynucleotide of any one of Embodiments 22 to 31, wherein the chimeric antigen receptor (CAR) comprises one or more antigen-binding domains.

[0233] Embodiment 33 is the isolated polynucleotide of any one of Embodiments 22 to 32, wherein the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0234] Embodiment 34 is a chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of any one of Embodiments 22 to 33.

[0235] Embodiment 35 is a vector comprising the isolated polynucleotide of any one of Embodiments 22 to 33.

[0236] Embodiment 36 is a host cell comprising the vector of embodiment 35.

[0237] Embodiment 37 is the host cell of embodiment 36, wherein the host cell is a T cell, preferably a human T cell.

[0238] Embodiment 38 is the host cell of embodiment 36, wherein the host cell is a NK cell, preferably a human NK cell.

[0239] Embodiment 39 is a method of producing a host cell that expresses a chimeric antigen receptor (CAR), the method comprising transducing a T cell with the vector of embodiment 35.

[0240] Embodiment 40 is a method for producing chimeric antigen receptor (CAR)-T cells, comprising culturing T cells comprising an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of Embodiments 22 to 33 under CAR-T cell-producing conditions, and recovering the CAR-T cells.

[0241] Embodiment 41 is a method of producing a host cell expressing a chimeric antigen receptor (CAR), comprising transducing an NK cell with the vector of embodiment 35.

[0242] Embodiment 42 is a method for producing chimeric antigen receptor (CAR)-NK cells, comprising culturing NK cells comprising an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of Embodiments 22 to 33 under CAR-NK cell-producing conditions, and recovering the CAR-NK cells.

[0243] Embodiment 43 is a method of generating a cell comprising a chimeric antigen receptor (CAR), comprising contacting a cell with an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of Embodiments 22 to 33, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

[0244] Embodiment 44 is a method of treating cancer in a subject in need thereof, comprising administering to a subject in need thereof the host cell of any one of Embodiments 36 to 38, optionally wherein the cancer is selected from lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML), and other liquid tumors.

[0245] Embodiment 45 is the method of embodiment 44, further comprising administering to a subject in need thereof an agent that increases the efficacy of the cells expressing the CAR.

[0246] Embodiment 46 is the method of embodiment 44, further comprising administering to a subject in need thereof an agent that ameliorates one or more side effects associated with administration of the cells expressing a CAR.

[0247] Embodiment 47 is the method of embodiment 44, further comprising administering to a subject in need thereof an agent for treating a GPC3-associated disease.

[0248] [Example] [Example 1] Identification of anti-TAA monoclonal antibodies Mice were immunized with the antigen, hybridomas were produced, and screened by ELISA and / or FACS. Positive clones were isolated and sequenced.

[0249] The sequences of the heavy and light chain variable regions (VH and VL regions, respectively) of anti-TAA monoclonal antibodies are provided in Tables 1 and 2, and the CDR regions of anti-TAA monoclonal antibodies are provided in Tables 3-6.

[0250] [Table 1-1] [Table 1-2]

[0251] [Table 2]

[0252] [Table 3-1] [Table 3-2]

[0253] [Table 4]

[0254] [Table 5]

[0255] [Table 6-1] [Table 6-2]

[0256] [Example 2] Production and purification of mAbs from the culture medium of transfected cells To obtain recombinant anti-TAA chimeric mAbs, expression vectors containing mouse variable regions (VH and VL) fused to the constant regions of human IgG1 heavy and kappa light chains, respectively, were transiently transfected into ExpiCHO-S or Expi293F cells. Recombinant antibodies produced in suspension in ExpiCHO-S or Expi293F cells were purified using Protein A affinity chromatography.

[0257] [Example 3] ELISA binding analysis of purified chimeric mAbs Purified chimeric mAbs were tested in an ELISA assay for their ability to bind to immobilized TIP-1. Recombinant human TIP-1 in PBS buffer was coated onto a 96-well plate for 1 hour at room temperature. The plate was blocked with 5% BSA in TBST for 1 hour at room temperature. Various concentrations of mAb were incubated in each well of each plate for 1 hour at room temperature. The plate was washed, and binding to TIP-1 was detected by incubating with anti-human IgG conjugated to horseradish peroxidase (hIgG-HRP) (ThermoFisher Scientific, Cat#: H10007) for 1 hour at room temperature. After washing, the ELISA was developed using One-Step Detection Solution (ThermoFisher Scientific, Cat#: 34028) and measured as absorbance at 450 nm. Anti-GPC3 chimeric antibodies were tested in a similar assay, except that nickel-coated plates (ThermoFisher, Cat#: 15442) were used. The results are shown in Figure 1 and Figures 2A to 2B.

[0258] [Example 4] FACS binding analysis of purified mAbs HEK293 cells expressing full-length human GPC3 were transferred to a 96-well plate. Approximately 50,000 cells were incubated with purified chimeric anti-GPC3 mAb (variable regions of mouse mAb fused to the constant regions of human IgG1 heavy and kappa light chains, respectively) for 15 minutes at 4°C. oThe cells were incubated at various concentrations in PBS (HBSS) at 100,000 cells per well. In some cases, 100,000 cells were used per well. The cells were then centrifuged for 5 minutes and washed once with FACS buffer (HBSS supplemented with 5% BSA and 0.05% sodium azide). The cells were then incubated with an Alexa Fluor 488-conjugated anti-human IgG secondary antibody (ThermoFisher, Cat#: H10120) and incubated on ice for an additional 15 minutes. The cells were then washed once in FACS buffer and resuspended in FACS buffer. The cells were then run on an Attune NxT column, and the data were analyzed using the Attune NxT software. The results are shown in Figures 2C-2E. HEK293 cells lacking GPC3 expression were used as a negative control (Figures 2F-2G).

[0259] [Example 5] Humanization of anti-GPC3 mAb A murine anti-GPC3 mAb was humanized to reduce the potential for immunogenicity when used in human patients. The sequences of the variable regions of the heavy and light chains (VH and VL) were compared with human antibody sequences in the Protein Data Bank (PDB) database to construct a homology model. The CDRs of both the heavy and light chains of the murine mAb were grafted onto a human framework most likely to maintain the proper structure required for antigen binding. Back-mutations or other mutations from human residues to murine residues were designed as necessary. The sequences of the humanized VH and VL regions are shown in Table 7. The humanized VH and VL regions were fused to the constant regions of human IgG1 heavy and kappa light chains, respectively. Binding of the humanized mAb to GPC3 was assessed using a FACS assay (Figures 3A–3O). M3-H1L1 refers to the humanized mAb constructed using the M3-H1 heavy and M3-L1 light chains shown in Table 7; other humanized clones follow the same naming convention.

[0260] [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5]

[0261] [Example 6] Construction of anti-GPC3 mAbs scFv molecules Humanized sequences of anti-GPC3 mAbs were used to construct scFv molecules fused to the human IgG1 Fc region. The sequences of the designed scFv molecules are listed in Table 8. The fusion molecules were expressed in CHO cells, purified, and tested for binding to GPC3 using a FACS assay (Figures 4A-4E).

[0262] [Table 8-1] [Table 8-2] [Table 8-3]

[0263] Those skilled in the art will appreciate that changes could be made to the above-described embodiments without departing from the broad inventive concept thereof. It is understood, therefore, that the invention is not limited to the particular embodiments disclosed, but that it is intended to cover modifications within the spirit and scope of the invention as defined herein.

Claims

1. An isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof, which specifically binds to GPC3, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 87, 88, 89, 90, 91, and 92, respectively, or SEQ ID NOs: 93, 94, 95, 96, 97, and 98, respectively.

2. 2. The isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof of claim 1, comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 85, 99-102, or 183-197, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 86, 103-105, or 198-201.

3. (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 101 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 102 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 105; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 100 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 188 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 103; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 189 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 190 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 195 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 194 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 104; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 187 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 199; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 196 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201, or (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201 2. The isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof of claim 1, comprising:

4. 4. The isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof of any one of claims 1 to 3, wherein the antibody or antigen-binding fragment or antigen-binding domain thereof is chimeric and / or human or humanized.

5. 4. The isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof according to any one of claims 1 to 3, wherein the antibody or antigen-binding fragment or antigen-binding domain thereof is capable of inducing effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or is capable of mediating the recruitment of conjugated drugs, and / or is capable of forming a bispecific antibody with another mAb or antigen-binding fragment thereof that has cancer-killing activity.

6. A bispecific antibody or antigen-binding fragment thereof or bispecific antigen-binding domain comprising the monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof according to any one of claims 1 to 5.

7. An isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof of any one of claims 1 to 5, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain thereof of claim 6.

8. A vector comprising the isolated nucleic acid of claim 7.

9. A host cell comprising the vector of claim 8.

10. A pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof according to any one of claims 1 to 5, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain thereof according to claim 6, and a pharmaceutically acceptable carrier.

11. Use of the isolated monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof according to any one of claims 1 to 5, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain thereof according to claim 6, in the preparation of a pharmaceutical composition or medicament for targeting GPC3 on the surface of cancer cells and / or treating cancer and / or treating an inflammatory disease in a subject in need thereof.

12. The use described in claim 11, wherein the cancer is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, bile duct cancer, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

13. A method for producing the monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof according to any one of claims 1 to 5, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain thereof according to claim 6, the method comprising culturing cells containing a nucleic acid encoding the monoclonal antibody or antigen-binding fragment or antigen-binding domain thereof, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain, under conditions for producing the monoclonal antibody or antigen-binding fragment or antigen-binding domain, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain, and recovering the monoclonal antibody or antigen-binding fragment or antigen-binding domain, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain from the cells or culture.

14. A method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment or antigen-binding domain of any one of claims 1 to 5, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain of claim 6, the method comprising combining the monoclonal antibody or antigen-binding fragment or antigen-binding domain, or the bispecific antibody or antigen-binding fragment or bispecific antigen-binding domain with a pharmaceutically acceptable carrier to obtain a pharmaceutical composition.

15. 1. A method for determining the level of GPC3 in a sample obtained from a subject, comprising: a. contacting a sample obtained from a subject with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 5; and b. Determining the level of GPC3 in a sample obtained from the subject A method comprising:

16. The method of claim 15, wherein the sample is a tissue sample or a blood sample.

17. The method described in claim 16, wherein the tissue sample is a cancer tissue sample.

18. 1. An isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an extracellular domain comprising at least one antigen-binding domain according to any one of claims 1 to 5 or a bispecific antigen-binding domain according to claim 6; (b) hinge region; (c) a transmembrane region, and (d) Intracellular signaling domain An isolated polynucleotide comprising:

19. 20. A chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of claim 18.

20. 20. A host cell comprising a vector comprising the isolated polynucleotide of claim 18.

21. The host cell described in claim 20, which is a T cell or an NK cell.

Images