Anti-trop2 SCFV and method of use thereof

Abstract

Disclosed are antigen binding molecules that specifically bind to the TROP2 extracellular domain. The antigen binding molecules include a single-chain variable fragment (scFv) and an antibody comprising a VH having a specific amino acid sequence and VL having a specific amino acid sequence. Diagnostic and therapeutic methods of using the same are also disclosed.

Description

[0001] ANTI-TROP2 SCFV AND METHOD OF USE THEREOF INCORPORATION OF SEQUENCE LISTING

[0002] This application contains a sequence listing submitted in Computer Readable Form (CRF). The CRF file contains the sequence listing entitled “1-2-PB8160001PCT-SequenceListing.xml”, which was created on October 2, 2025, and is 96,948 bytes in size. The information in the sequence listing is incorporated herein by reference in its entirety.

[0003] FIELD OF THE INVENTION

[0004] The present invention relates generally to the preparation and use of an anti-human TROP2 antibody or single-chain variable fragment (scFv) for cancer therapy.

[0005] BACKGROUND OF THE INVENTION

[0006] Antibody-based cancer therapy has become one of the most promising cancer treatments. It utilizes antibodies, a type of protein naturally produced by the immune system, to target and destroy cancer cells. This approach leverages the specificity of antibodies to recognize and bind to certain proteins (also called antigens or cancer targets) usually on the surface of cancer cells or in the extracellular space, utilizing various strategies to attack those cells while minimizing damage to normal cells.

[0007] As a candidate for cancer targets, trophoblast cell surface antigen 2 (TROP2) is a 35-kDa transmembrane, signal transducing type-1 glycoprotein related to tumor marker EpCAM (Lenart, S. et al. Trop2: Jack of All Trades, Master of None. Cancers 12, 3328 (2020)). TROP2 was first identified in 1981 on human trophoblast cells, and its overexpression was found to promote cancer growth in vitro and in vivo. Elevated TROP2 expressions were detected in most epithelial neoplasms (urothelial, salivary gland, ovarian, endometrial, gastric, oral squamous cell, lung, cervical, breast and colorectal cancer) and have been associated with poor cancer patients’ prognosis (Zeng, P et al. Impact of TROP2 expression on prognosis in solid tumors: A Systematic Review and Meta-analysis. Sci Rep 6, 33658 (2016)).

[0008] Recent clinical data validated TROP2 as a candidate for targeted therapy with antibodydrug conjugates showing potent anti-tumor activity and mild, reversible toxicity. Antibody-drug conjugate (ADC) sacituzumab govitecan (Trodelvy), composed of a humanized anti-TROP2 (hRS7) monoclonal antibody and topoisomerase I inhibitor SN-38, achieved response rates ranging from 14% - 19% in small- and non-small-cell lung cancer, to ~30% in urothelial and breast cancer (Goldenberg, D. M. & Sharkey, R. M Sacituzumab govitecan, a novel, third-generation, antibody-drug conjugate (ADC) for cancer therapy. Expert Opinion on Biological Therapy 20, 871-885 (2020)). These favorable outcomes established the basis for accelerated approval of sacituzumab govitecan in relapsed metastatic triple-negative breast cancer and fast-track designation in the treatment of recurrent urothelial cancer.

[0009] TROP2 as a cancer therapeutic target presents a multitude of opportunities for therapeutics development including but not limited to monoclonal antibody, ADC, T cell engager (TCE), and immunotoxin (Goldenberg, D. M., Stein, R. & Sharkey, R. M. The emergence of trophoblast cell-surface antigen 2 (TROP-2) as a novel cancer target. Oncotarget 9, 28989-29006 (2018)). A critical component of these therapeutics is a highly specific and selective anti-TROP2, fully human monoclonal antibody.

[0010] As there is a strong need to develop anti-TROP2 therapeutical agents, the present invention describes the discovery' of a group of anti-TROP2 molecules that are in the format of scFvs and validated as IgGs.

[0011] SUMMARY OF THE INVENTION

[0012] The instant disclosure provides an antigen binding protein molecule that specifically binds to the human TROP2 extracellular domain.

[0013] Said antigen binding protein molecule is selected from the group consisting of an antibody and an antigen binding fragment. Examples of the antigen binding fragment include a single-chain variable fragment (scFv), a Fab' (fragment antigen-binding, monomeric), a F(ab')2 (Fragment antigen-binding, dimeric), a single chain diabody, and a nanobody.

[0014] Specifically, said antigen binding protein molecule is an scFv. More specifically, in one embodiment, the scFv can be selected from the group consisting of an scFv comprising VH having the amino acid of SEQ ID NO: 1 and VL having the amino acid of SEQ ID NO: 2, an scFv comprising VH having the amino acid of SEQ ID NO: 3 and VL having the amino acid of SEQ ID NO: 4, an scFv comprising VH having the amino acid of SEQ ID NO: 5 and VL having the amino acid of SEQ ID NO: 6, an scFv comprising VH having the amino acid of SEQ ID NO: 7 and VL having the amino acid of SEQ ID NO: 8, an scFv comprising VH having the amino acid of SEQ ID NO: 9 and VL having the amino acid of SEQ ID NO: 10, an scFv comprising VH having the amino acid of SEQ ID NO: 11 and XT, having the amino acid of SEQ ID NO: 12, an scFv comprising VH having the amino acid of SEQ ID NO: 13 and VL having the amino acid of SEQ ID NO: 14, an scFv comprising VH having the amino acid of SEQ ID NO: 15 and VL having the amino acid of SEQ ID NO: 16, and an scFv comprising VH having the amino acid of SEQ ID NO: 17 and VL having the amino acid of SEQ ID NO: 18.

[0015] In another embodiment, the scFv can be selected from the group consisting of an scFv comprising the amino acid sequence of SEQ ID NO: 19, an scFv comprising the amino acid sequence of SEQ ID NO: 20, an scFv comprising the amino acid sequence of SEQ ID NO: 21, an scFv comprising the amino acid sequence of SEQ ID NO: 22, an scFv comprising the amino acid sequence of SEQ ID NO: 23, an scFv comprising the amino acid sequence of SEQ ID NO: 24, an scFv comprising the amino acid sequence of SEQ ID NO: 25, an scFv comprising the amino acid sequence of SEQ ID NO: 26, and an scFv comprising the amino acid sequence of SEQ ID NO: 27.

[0016] In yet another embodiment, the scFv can be selected from the group consisting of an scFv comprising VII having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 28, 29, and 30, respectively; an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 34, 35, and 36, respectively; an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 40, 41, and 42, respectively; an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 46, 47, and 48, respectively; an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively; an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 58, 59, and 60, respectively; an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 64, 65, and 66, respectively; an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 70, 71, and 72, respectively; and an scFv comprising VH having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 76, 77, and 78, respectively.

[0017] Alternatively, the scFv can be selected from the group consisting of an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 31, 32, and 33, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 37, 38, and 39, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 43, 44, and 45, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 49, 50, and 51, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 55, 56, and 57, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 61, 62, and 63, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 67, 68, and 69, respectively; an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively; and an scFv comprising VL having the CDRI, CDR2, and CDR3 sequences of SEQ ID NOs: 79, 80, and 81, respectively.

[0018] Equally advantageously, said antigen binding protein molecule can be an antibody. Preferably, the antibody can be a humanized antibody. In one embodiment, the antibody can be selected from the group consisting of an antibody comprising VII having the amino acid of SEQ ID NO: 1 and VL having the amino acid of SEQ ID NO: 2, an antibody comprising VH having the amino acid of SEQ ID NO: 3 and VL having the amino acid of SEQ ID NO: 4, an antibody comprising VH having the amino acid of SEQ ID NO: 5 and VL having the amino acid of SEQ ID NO: 6, an antibody comprising VH having the amino acid of SEQ ID NO: 7 and VL having the amino acid of SEQ ID NO: 8, an antibody comprising VH having the amino acid of SEQ ID NO: 9 and VL having the amino acid of SEQ ID NO: 10, an antibody comprising VH having the amino acid of SEQ ID NO: 11 and VL having the amino acid of SEQ ID NO: 12, an antibody comprising VH having the amino acid of SEQ ID NO: 13 and VL having the amino acid of SEQ ID NO: 14, an antibody comprising VH having the amino acid of SEQ ID NO: 15 and VL having the amino acid of SEQ ID NO: 16, and an antibody comprising VH having the amino acid of SEQ ID NO: 17 and VL having the amino acid of SEQ ID NO: 18.

[0019] In another embodiment, the antibody can be selected from the group consisting of an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 28, 29, and 30, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 34, 35, and 36, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 40, 41, and 42, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 46, 47, and 48, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 58, 59, and 60, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 64, 65, and 66, respectively; an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 70, 71, and 72, respectively; and an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 76, 77, and 78, respectively.

[0020] Alternatively, the antibody is selected from the group consisting of an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 31, 32, and 33, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 37, 38, and 39, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 43, 44, and 45, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 49, 50, and 51, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 55, 56, and 57, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 61, 62, and 63, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 67, 68, and 69, respectively; an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively; and an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 79, 80, and 81, respectively.

[0021] Another aspect of the present invention relates to a composition of an scFv or antibody that comprises the amino acid sequence representing such molecules.

[0022] In one embodiment, an scFv or antibody that specifically binds to TROP2 comprises: a VH CDR1 having the amino acid of SEQ ID NO: 28, a VH CDR2 having the amino acid of SEQ ID NO: 29, a VH CDR3 having the amino acid of SEQ ID NO: 30, a VL CDR1 having the amino acid of SEQ ID NO: 31, a VL CDR2 having the amino acid of SEQ ID NO: 32, and a VL CDR3 having the amino acid of SEQ ID NO: 33.

[0023] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 1. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 2.

[0024] In another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDR1 having the amino acid of SEQ ID NO: 34, a VH CDR2 having the amino acid of SEQ ID NO: 35, a VH CDR3 having the amino acid of SEQ ID NO: 36, a VL CDR1 having the amino acid of SEQ ID NO: 37, a VL CDR2 having the amino acid of SEQ ID NO: 38, and a VL CDR3 having the amino acid of SEQ ID NO: 39.

[0025] The scFv or antibody that is immediately proceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 3. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 4.

[0026] In still another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDR1 having the amino acid of SEQ ID NO: 40, a VH CDR2 having the amino acid of SEQ ID NO: 41, a VH CDR3 having the amino acid of SEQ ID NO: 42, a VL CDR1 having the amino acid of SEQ ID NO: 43, a VL CDR2 having the amino acid of SEQ ID NO: 44, and a VL CDR3 having the amino acid of SEQ ID NO: 45.

[0027] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 5. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 6.

[0028] In yet another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDR1 having the amino acid of SEQ ID NO: 46, a VH CDR2 having the amino acid of SEQ ID NO: 47, a VH CDR3 having the amino acid of SEQ ID NO: 48, a VL CDR1 having the amino acid of SEQ ID NO: 49, a VL CDR2 having the amino acid of SEQ ID NO: 50, and a VL CDR3 having the amino acid of SEQ ID NO: 51.

[0029] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 7. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 8.

[0030] In another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDR1 having the amino acid of SEQ ID NO: 52, a VH CDR2 having the amino acid of SEQ ID NO: 53, a VH CDR3 having the amino acid of SEQ ID NO: 54, a VL CDR1 having the amino acid of SEQ ID NO: 55, a VL CDR2 having the amino acid of SEQ ID NO: 56, and a VL CDR3 having the amino acid of SEQ ID NO: 57.

[0031] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 9. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 10. In still another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDR1 having the amino acid of SEQ ID NO: 58, a VH CDR2 having the amino acid of SEQ ID NO: 59, a VH CDR3 having the amino acid of SEQ ID NO: 60, a VL CDRl having the amino acid of SEQ ID NO: 61, a VL CDR2 having the amino acid of SEQ ID NO: 62, and a VL CDR3 having the amino acid of SEQ ID NO: 63.

[0032] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 11. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 12.

[0033] In another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDRl having the amino acid of SEQ ID NO: 64, a VH CDR2 having the amino acid of SEQ ID NO: 65, a VH CDR3 having the amino acid of SEQ ID NO: 66, a VL CDRl having the amino acid of SEQ ID NO: 67, a VL CDR2 having the amino acid of SEQ ID NO: 68, and a VL CDR3 having the amino acid of SEQ ID NO: 69.

[0034] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 13. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 14.

[0035] In yet another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDRl having the amino acid of SEQ ID NO: 70, a VH CDR2 having the amino acid of SEQ ID NO: 71, a VH CDR3 having the amino acid of SEQ ID NO: 72, a VL CDRl having the amino acid of SEQ ID NO: 73, a VL CDR2 having the amino acid of SEQ ID NO: 74, and a VL CDR3 having the amino acid of SEQ ID NO: 75.

[0036] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 15. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 16.

[0037] In still another embodiment, a scFv or antibody that specifically binds to TROP2 comprises a VH CDR1 having the amino acid of SEQ ID NO: 76, a VH CDR2 having the amino acid of SEQ ID NO: 77, a VH CDR3 having the amino acid of SEQ ID NO: 78, a VL CDR1 having the amino acid of SEQ ID NO: 79, a VL CDR2 having the amino acid of SEQ ID NO: 80, and a VL CDR3 having the amino acid of SEQ ID NO: 81,

[0038] The scFv or antibody that is immediately preceding can also have a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 17. Alternatively, it can also have a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 18.

[0039] The scFv molecule described above comprises a linker between their VH and VL, and is capable of specifically binding to human TROP2. On the other hand, the antibody described above contains a variable region and a constant region and is capable of specifically binding to human TROP2 The antibody can be preferably a humanized antibody.

[0040] In another aspect, this invention provides a nucleic acid encoding the antigen binding protein molecule described above, which includes an scFv or an antibody, and a vector comprising said nucleic acid. The nucleic acid encodes the VH and VL responsible for binding to human TROP2. The vector can be used for expression in a host cell for therapeutic or diagnostic purposes.

[0041] Also provided is a cell comprising said vector and methods using the same. The cell can be selected from a mammalian cell, bacterial cell, yeast cell, or insect cell, and is capable of expressing the antigen binding protein molecule as above discussed.

[0042] One of the methods is for making an antigen binding molecule as described above. The method comprises culturing said cell under suitable conditions. Another method is for a cellbased therapy, which comprises administering to a subject in need thereof a preselected number of said cells expressing a therapeutic dose of the above-described antigen binding protein molecule.

[0043] Still another aspect of this invention relates to a pharmaceutical composition of the antigen binding protein molecule introduced above and methods using the same. Specifically, one method is for administering a dose of a medicament, which comprises administering to a subject in need thereof an effective dose of said pharmaceutical composition. Another method is for treating a disease, which comprises administering to a subject in need thereof an effective amount of said pharmaceutical composition. The administration can be carried out by a delivery method selected from the group consisting of intravenous, subcutaneous, or intraperitoneal injection. The subject can be a human.

[0044] Yet another method is for treating cancer, which also comprises administering to a subject in need thereof an effective amount of said pharmaceutical composition. The cancer can be selected from the group consisting of epithelial cancers, breast cancer, lung cancer, ovarian cancer, colorectal cancer, pancreatic cancer, and prostate cancer. Advantageously, the composition can be administered via intravenous or subcutaneous injection.

[0045] Further methods using said pharmaceutical composition include that for inhibiting or suppressing a tumor in a subject and that for delaying the progression of a solid tumor in a subject. Both require administering an effective amount of said pharmaceutical composition. Further, the solid tumor can be selected from the group consisting of epithelial cancers, breast cancer, lung cancer, ovarian cancer, colorectal cancer, pancreatic cancer, and gastric cancer.

[0046] In yet another aspect, this invention provides an antigen binding protein molecule, as described above, further with a detectable label. Examples of the detectable label include a fluorescent label, a photochromic compound, a radiolabel, or a hapten.

[0047] Said antigen binding protein molecule with a detectable label can be applied to a method for detecting the presence of human TROP2 in a sample, which comprises contacting the sample with said antigen binding protein molecule and a positive measurement of the detectable label can indicate the presence of human TROP2 in the sample. In additional, the sample can be selected from the group consisting of blood, serum, tissue biopsy, or cell lysate

[0048] Said antigen binding protein molecule can also be applied to a method for quantifying the amount of human TROP2 in a sample, which comprises contacting the sample with said antigen binding protein molecule of claim 54 and a numeric measurement of the detectable label can indicate the amount of human TROP2 in the sample. Said antigen binding protein molecule can be further applied to a method of diagnosing the presence of a tumor or a cancer growth through a clinical sample, which includes: (a) obtaining a clinical sample isolated from a patient in need thereof, wherein the clinical sample can be selected from the group consisting of blood, serum, tissue biopsy, or cell lysate; and (b) contacting the clinical sample with said antigen binding protein molecule. The resulting positive measurement of the detectable label is indicative of the presence of a tumor or cancer growth associated with TROP2 expression in the patient. The antigen binding protein molecule with a detectable label as described above can be studied for its biodistribution profile as determined by pharmacokinetics measurements. It can also be determined whether it is suitable for whole body imaging and utilization as a biomarker for assessing human TROP2 organ distribution.

[0049] Consequently, a method to use the antigen binding protein molecule discussed immediately above is for diagnosing the presence of a tumor or a cancer growth in a subject through in vivo imaging, which comprises: (a) administering to the subject an effective amount of a composition comprising said antigen binding protein molecule, wherein the detectable label is suitable for the in vivo imaging; and (b) after a period of time sufficient for said antigen binding protein molecule to diffuse, applying whole-body or regionalized imaging to detect the bound antigen binding protein molecule by the detectable label; whereby a positive measurement of the detectable label indicates the presence of a tumor or cancer growth associated with TROP2 expression in the subject.

[0050] This invention also provides a kit for detecting human TROP2 and a kit for quantifying the amount of human TROP2 in a sample. Both kits comprise said antigen binding protein molecule.

[0051] The details of the invention are set forth in the drawing and the description below. Other features, objects, and advantages of the invention will be apparent to those persons skilled in the art upon reading the drawing and the description, as well as from the appended claims.

[0052] BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The embodiments exemplified in Figures 1-6 are intended to illustrate the present invention and are not to be considered as limiting.

[0054] The accompanying drawings are not intended to be drawn to scale. The figures are illustrative only and are not required for enablement of the disclosure. For purposes of clarity, not every component may be labeled in every drawing.

[0055] Figure 1 is a plot of an assay in which crude supernatant from 293T cells transfected with scFv-Fc clones was used in single point ELISA screening against recombinant TROP-2 protein. The ELISA signals were represented by the bars and the measured OD600 numbers were represented by the left Y-axis. The dots connected by a line represent the expression titers corresponding to the numbers on the right Y-axis. Clone numbers are indicated on the X-axis.

[0056] Figure 2 is a series of flow cytometry analysis plots with fluorescence-activated cell sorting (F / ACS) showing none of the 21 ELISA-positive clones bound to 293T cells in the FACS analysis. Figure 3 is a series of FACS plots showing that the TROP-2 negative cell line was used in FACS analysis against the ELISA positive clones. Clones T1, T6, and T23 displayed different levels of non-specific binding, whereas the others didn’t.

[0057] Figure 4 is a series of FACS plots showing that 293T transiently transfected and expressing a high level of TROP-2 was used in the FACS assay. Positive control is a rabbit polyclonal antibody. Almost all the clones displayed various binding activities against the TROP-2 high expressing 293T cell.

[0058] Figure 5 is a series of FACS plots showing that prostate cancer cell line PC3 was used in the FACS analysis. Indicated clones showing binding to 293T also bound PC3.

[0059] Figure 6 is a plot of an assay in which serial dilutions of scFv-Fc of indicated clones were added to TROP-2 recombinant protein coated on an ELISA plate and the bindings were measured and ECso calculated.

[0060] DETAILED DESCRIPTION

[0061] Before the present invention is described in more detail, it is to be understood that this invention is not limited to a particular method described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0062] The present disclosure focuses on antibodies and antigen-binding fragments thereof. Antigen-binding fragments (Fab) are portions of antibodies responsible for recognizing and binding to specific antigens. Various types of antibody binding motifs have been developed, which include Fab' (Fragment antigen-binding, monomeric), F(ab')2 (Fragment antigen-binding, dimeric), scFv, and single-domain antibody (VHH or nanobody ).

[0063] A scFv consists of the variable regions of the heavy (VH) and light (VL) chains of an antibody, joined by a short peptide linker. These fragments are small and maintain the antigenbinding properties of the full antibody. Due to its size and flexibility, the scFv is widely used in therapeutic applications.

[0064] The antibody of the present invention can also be a fully human monoclonal antibody. Fully human monoclonal antibodi es can be generated by any number of techniques with which those having ordinary skill in the art will be familiar. Such methods include, but are not limited to, Epstein Barr Virus (EBV) transformation of human peripheral blood cells, in vitro immunization of human B-cells, a fusion of spleen cells from immunized transgenic mice carrying inserted human immunoglobulin genes, isolation from human immunoglobulin V region phage libraries, or other procedures as known in the art. As used herein, the terms "antigen" and "epitope" are well understood in the art and refer to the portion of a macromolecule (e.g., a polypeptide) which is specifically recognized by a component of the immune system, e.g., an antibody or a T-cell antigen receptor.?\s used herein, the term "antigen" encompasses antigenic epitopes, e.g., fragments of an antigen which are antigenic epitopes. Epitopes can be recognized by antibodies in solution, e.g. free from other molecules. Epitopes can be recognized by T-cell antigen receptors when the epitope is associated with a class I or class II major histocompatibility complex molecule.

[0065] The term “tumor-associated antigen” or “TAA” refers to an antigen that is associated with a tumor.

[0066] As used herein, the terms "subject," "host," "patient," and "individual" are used interchangeably herein to refer to any mammalian subject for whom diagnosis or therapy is desired, particularly humans. Other subjects may include cattle, dogs, cats, guinea pigs, rabbits, rats, mice, horses, and so on.

[0067] The terms "cell," and "cells," and "cell population," used interchangeably, intend one or more mammalian cells. The term includes progeny of a cell or cell population. Those skilled in the art will recognize that "cells" include progeny of a single cell, and there are variations between the progeny and its original parent cell due to natural, accidental, or deliberate mutation and / or change.

[0068] The terms "cell proliferation" and "to proliferate" as used herein refer to the amplification of the cell-by-cell division.

[0069] A "cancer cell" as used herein refers to a cell exhibiting a neoplastic cellular phenotype, which may be characterized by one or more of, for example, abnormal cell growth, abnormal cellular proliferation, loss of density dependent growth inhibition, anchorage-independent growth potential, ability to promote tumor growth and / or development in an immunocompromised non-human animal model, and / or any appropriate indicator of cellular transformation. "Cancer cell" may be used interchangeably herein with "tumor cell" or "cancerous cell", and encompasses cancer cells of a solid tumor, a semi-solid tumor, a primarytumor, a metastatic tumor, and the like.

[0070] As used herein, "immunotherapy" refers to the treatment of a disease (e.g., cancer) by modulating an immune response to a disease antigen. In the context of the present application, immunotherapy refers to providing an anti -cancer immune response in a subject by administration of an antibody (e.g., a monoclonal antibody) and / or by administration of an antigen that elicits an anti -tumor antigen immune response in the subject. The term “immunoglobulin” means an immune molecule from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG, and IgM IgG subclasses are also well known to those in the art and include but are not limited to human IgGl, IgG2, IgG3, and IgG4. Many of the molecules described herein are immunoglobulins. As used herein, “isotype” means the antibody class or subclass (e.g., IgM or IgGl) that is encoded by the heavy chain constant region genes.

[0071] The term “heavy chain” when used in reference to an antibody can refer to any distinct type, e.g., alpha (a), delta (5), epsilon (s), gamma (y), and mu (p), based on the amino acid sequence of the constant domain, which give rise to IgA, IgD, IgE, IgG and IgM cl asses of antibodies, respectively, including subclasses of IgG, e.g., IgGl, IgG2, IgG3, and IgG4.

[0072] On the other hand, the term “light chain” when used in reference to an antibody can refer to any distinct type, e.g., kappa (K) or lambda (X) based on the amino acid sequence of the constant domains. Light chain amino acid sequences are known in the art. In specific embodiments, the light chain is a human light chain.

[0073] The terms “CDR” or “complementarity determining region” refer to amino acid residues comprising non-contiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. The amino acids of the CDRs of the variable domains were initially defined by Kabat (Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991)). Based on sequence variability, the CDRs include amino acid sequences of CDR1, CDR2, and CDR3 in the heavy chain variable domain (VH) and CDR1, CDR2, and CDR3 in the light chain variable domain (VL). The CDR sequences and their three-dimensional structural loops formed were found to be important in antigen binding activity.

[0074] The term “antigen presenting cells (APCs)” refers to a class of cells capable of presenting one or more antigens in the form of peptide-WIC complex recognizable by specific effector cells of the immune system, and thereby inducing an effective cellular immune response against the antigen or antigens being presented.

[0075] “Immune response” broadly refers to the antigen-specific responses of lymphocytes to foreign substances. Any substance that can elicit an immune response is said to be “immunogenic” and is referred to as an “immunogen”. All immunogens are antigens, however, not all antigens are immunogenic. An immune response of this invention can be humoral (via antibody activity) or cell-mediated (via T cell activation).

[0076] The term “binding affinity” means the strength of the total of non-covalent interactions between a single binding site of a molecule (e.g., an antigen binding fragment or an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its partner Y can generally be represented by the dissociation constant (KD). Affinity can be measured and / or expressed in a number of ways known in the art, including, but not limited to, equilibrium dissociation constant (KD), and equilibrium association constant (KA).

[0077] An "effective amount" is an amount sufficient to effect beneficial or desired clinical results. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of reagent antibodies is an amount that is suffici ent to diagnose, palliate, ameliorate, stabilize, reverse, slow, or delay the progression of the disease state.

[0078] The term "ECso" (half-maximal effective concentration) is an important parameter that measures the potency of a drug or antibody. It represents the concentration required to achieve 50% of the maximum effect. A lower ECso value indicates a higher affinity of the drug or antibody for the target, meaning it can achieve half of its maximum effect at a lower concentration. In antibody screening, a lower ECso suggests stronger binding to the target antigen.

[0079] "PC3" is a prostate cancer cell line commonly used in cancer research. PC3 cells express low levels of TROP-2 protein, making them useful in antibody screening or functional studies to evaluate the binding ability of antibodies in a low TROP-2 expression context. If an antibody binds effectively to PC3 cells, it indicates potential therapeutic relevance for cells with low TROP-2 expression.

[0080] "NCI-H1975" is a non-small cell lung cancer (NSCLC) cell line. In this case, if it is described as a TROP-2 high expression line, it indicates that TROP-2 is overexpressed on the surface of these lung cancer cells, making it a suitable model for testing antibodies or therapies targeting TROP-2 in lung cancer.

[0081] "NCI-H929" is a multiple myeloma cell line. If it is described as having high TROP-2 expression, it implies that TROP-2 is abundantly present on these multiple myeloma cells. This is notable because TROP-2 is more commonly associated with epithelial tumors, so its high expression in a hematologic cancer like multiple myeloma suggests that it could also serve as a target in these cancers.

[0082] "FACS" (Fluorescence-Activated Cell Sorting) is a specialized type of flow cytometry used for sorting and analyzing cells based on specific characteristics, such as size, shape, and the presence of fluorescently labeled markers. A "single-point ELISA" (Enzyme-Linked Immunosorbent Assay) refers to a simplified version of the traditional ELISA, where the concentration of an analyte (e.g., a protein or antibody) is determined based on one specific point or dilution of the sample, rather than creating a full standard curve.

[0083] The immunotherapeutic strategy, introduced in this invention, leverages the specificity of a scFv or antibody in targeting cancer antigens. The scFvs or antibodies may or may not compete with one another for binding to that antigen. In practice, one of the scFvs or antibodies provided herein is prepared in a pharmaceutical composition. Alternatively, two or more scFvs or antibodies provided herein are combined together in a pharmaceutical composition.

[0084] Preferably such a composition will be suitable for administration to a subject, including a human.

[0085] BEST MODE FOR CARRYING OUT INVENTION

[0086] The following example explains the present invention more concretely, but does not limit the range of the present invention.

[0087] EXAMPLES

[0088] Example 1:

[0089] Phage display strategy was applied in the discovery effort. The fully human, scFv phage library is the proprietary JechoMab platform established within Jecho Laboratories. The panning antigen his-tagged TROP2 extracellular domain was purchased from Sino Biological (#10428-H08H). The panning antigen was biotin-labeled and used in the magnetic bead-based (Dynabeads™ M-280 Streptavidin, Invitrogen, cat # 11205D) solution panning procedure. After three rounds of panning, single colonies were picked and subjected to monoclonal phage ELISA screening. Positive clones were picked and sequenced.

[0090] After sequencing, 30 unique sequences were cloned into the scFv-Fc expression vector and transfected into 293 T cells for transient expression. The resulting transfection supernatant was used in a single-point ELISA to select positive binders. Among the 30 scFv-Fc, 27 were successfully expressed and 21 displayed positive binding to TROP-2 protein in an ELISA as shown in Figure 1. The ELISA signals were represented by the bars and the measured OD600 numbers were represented by the left Y-axis. The dots connected by a line represent the expression titers corresponding to the numbers on the right Y-axis. Clone numbers are indicated on the X-axis.

[0091] Clones T4, T15, T17, T18, T19, T20, T22, T25, T26, and T28: Show very low bar heights, indicating weak binding or response. However, T17 shows a higher titer value, indicating that T7 was successfully expressed.

[0092] Clones Tl, T2, T3, T5, T6, T7, T8, T9, T10, TH, T12, T13, T14, T16, T21, T23, T24, T27, T29, and T30: Display high bar heights, suggesting strong binding or a higher response level. Figure 1 compares the binding response (bar graph) with another metric (line graph) for each antibody clone. 21 Clones seem to perform better, making them suitable candidates for further development.

[0093] Example 2:

[0094] The 21 clones that were positive in the preliminary ELISA were further characterized against cell lines. First, they were tested in FACS against cell lines negative for TROP-2 expressions including 293T and NCI-H929. None of the clones bound 293T which is the negative for TROP-2 expression as shown in Figure 2.

[0095] Figure 2 shows the fluorescence intensity distribution obtained through flow cytometry (FACS) analysis of multiple antibody clones binding on cells. Each plot presents the binding data of different antibody clones (e.g., TIC, T2C) compared to a control group (control).

[0096] X-axis in Figure 2: Represents fluorescence intensity; further to the right, the stronger the fluorescence signal indicating antibody binding to the cells.

[0097] Y-axis in Figure 2: Represents the proportion of cells, showing the distribution of cells at different fluorescence intensities.

[0098] Peak of the curve in Figure 2: Represents the fluorescence intensity of the cell population. The further to the right the peak is, the stronger the antibody binds to the cells, the further to the left, the weaker or no binding.

[0099] Control group (control, blue curve): This represents the background fluorescence of cells without antibody binding.

[0100] The left plot on Figure 2: Shows the FACS results for clones TIC to T8C. The curves for these clones are very close to the control group (blue), indicating weak or no binding to the target cells. The middle plot on Figure 2: Shows the FACS results for clones T9C to T16C. Similarly, these clones have curves very' close to the control, suggesting weak or no significant binding to the cells. The right plot on Figure 2: Shows the FACS results for clones T17C to T30C. Most of these clones also show little difference from the control group, indicating weak binding to the cells.

[0101] From Figure 2, it can be observed that most antibody clones show very weak binding to 297T which is the negative for TROP-2 expression, almost identical to the control group, indicating that these clones may exhibit specific binding to the target protein TROP-2. These data can be used for further screening and optimization of antibody clones.

[0102] Example 3: However, Figure 3 presents that some of the clones demonstrated bindings to the TROP-2 negative multiple myeloma cancer cell line NCI-H929. Clones Tl, T6, and T23 displayed different levels of non-specific binding, whereas the others didn’t.

[0103] Figure 3 presents flow cytometry' (FACS) results comparing the binding activity of different antibody clones (labeled H929-T1 through H929-T30) against the H929 multiple myeloma cell line, which is TROP-2 negative.

[0104] Interpretation:

[0105] Top Left Plot on Figure 3:

[0106] This plot shows the FACS data for clones H929-T1 through H929-T7 compared with the H929 control (blue line).

[0107] The curves for most clones (except H929-T1 and H929-T6) closely overlap with the control, indicating weak or no significant binding to the H929 cells.

[0108] Top Right Plot on Figure 3:

[0109] The binding of clones H929-T8 through H929-T14 is displayed here. Most of the clones show curves that align closely with the control (H929 control, blue), indicating weak or no binding.

[0110] None of these clones show a significant binding signal in this assay.

[0111] Bottom Left Plot on Figure 3:

[0112] This plot displays the FACS data for clones H929-T16 through H929-T24.

[0113] The H929-T23 clone (yellow line) shows a significant deviation, suggesting stronger non-specific binding to the TROP-2 negative H929 cells. Other clones (H929-T16, H929-T17) show limited or no binding, as their curves largely follow the control line.

[0114] Bottom Right Plot on Figure 3:

[0115] This final plot shows the binding of H929-T2, H929-T3, H929-T7, H929-T12, H929-T21, H929-T29, and H929-T30 compared with both the H929 control and H929 with a known TROP-2 antibody (purple line).

[0116] The purple line, which represents the H929 cells treated with a known TROP-2 antibody, serves as a positive control to indicate potential non-specific binding or cross-reactivity. The purple curve shifts significantly to the right, indicating strong binding, while the other clones largely overlap with the H929 control, suggesting no significant specific binding to the TROP-2 negative H929 cells.

[0117] Across all plots, most of the antibody clones tested show weak or no specific binding to the TROP-2 negative H929 cell line, as their curves closely follow the control. However, a few clones, such as H929-T1, T6 (in the top-left plot) and H929-T23 (in the bottom-left plot), show some non-specific binding. The purple line in the bottom-right plot demonstrates what strong binding would look like, serving as a positive control with TROP-2 specific antibody.

[0118] This analysis suggests that most of the tested clones are not binding specifically to H929 cells, which could be expected given the absence of TROP-2 on these.

[0119] Example 4:

[0120] These clones bound with various strengths to 293 T transiently transfected with TROP-2, as shown in Figure 4.

[0121] Figure 4 shows the binding profiles of various antibody clones to 293T cells transfected with TROP-2, which is demonstrated through flow cytometry (FACS). Each graph represents the fluorescence intensity (indicative of antibody binding) of different clones, compared to each other.

[0122] X-axis on Figure 4: Represents fluorescence intensity, indicating the binding strength of the antibody clones to TROP-2 expressing 293T cells. The further to the right the curve shifts, the stronger the binding.

[0123] Y-axis on Figure 4: Represents the percentage of the maximum cell population displaying a specific fluorescence intensity, showing the distribution of binding levels.

[0124] Different clones (color-coded) on Figure 4: Each colored line corresponds to a specific antibody clone, showing how it interacts with the TROP-2 expressing cells.

[0125] The red, yellow, and green lines represent different antibody clones. Some clones show a stronger shift to the right (higher binding intensity), indicating better binding to the TROP-2 positive 293T cells, while others are closer to the left, suggesting weaker binding.

[0126] In some graphs, clones demonstrate more prominent peaks than others, which signifies that certain clones may have a higher affinity for TROP-2, resulting in more effective binding to the transfected 293T cells.

[0127] Overall, this figure demonstrates all the clones displayed various binding activities against the TROP-2 high-expressing 293 T cell.

[0128] Example 5:

[0129] Figure 5 presents that selected clones are also bound to PC3 cells expressing low levels of TROP-2.

[0130] Figure 5 shows flow cytometry (FACS) results analyzing the binding of different TROP-2 antibody clones to TROP-2 expressing cells. The data is displayed in several panels, each comparing the fluorescence intensity (X-axis, RL1-A) and the percentage of maximum cell population (Y-axis, Percent of Max) for different antibody clones. X-axis on Figure 5: Represents fluorescence intensity, which indicates the strength of the antibody binding to TROP-2 on the surface of the cells. Further to the right a peak is, the stronger the antibody is binding.

[0131] Y-axis on Figure 5: Shows the percentage of the cell population that exhibits a particular fluorescence intensity, indicating the distribution of antibody binding across the cell population.

[0132] Different colored lines: Each panel contains curves for various clones (e.g., trop2-2, trop2-3, etc.) along with a known TROP-2 antibody (yellow, trop2+AB), which serves as a positive control for strong binding to TROP-2.

[0133] Top Left Panel on Figure 5:

[0134] This compares the control (trop2 control 1) with the known TROP-2 antibody (trop2+AB).

[0135] The yellow curve is shifted significantly to the right, indicating strong TROP-2 binding, while the red curve (control) shows no binding.

[0136] Top Right Panel on Figure 5:

[0137] This compares clones trop2-7, trop2-12, and trop2-21 with the positive control.

[0138] The positive control (yellow) shows strong binding, while trop2-7 and trop2-12 show moderate binding (curves shifted slightly right compared to the control), and trop2-21 shows weaker binding.

[0139] Bottom Left Panel on Figure 5:

[0140] This compares clones trop2-2, trop2-3, trop2-5 with the positive control.

[0141] The positive control (yellow) shows strong binding, while the other clones show various levels of binding. TROP2-5 has the weakest binding, indicated by the smaller right shift.

[0142] Bottom Right Panel on Figure 5:

[0143] This compares clones trop2-27, trop2-29, trop2-30 with the positive control.

[0144] The positive control again shows strong binding, while trop2-29 and trop2-30 show moderate binding levels, with trop2-27 showing weaker binding.

[0145] In each panel, the known TROP-2 antibody (yellow curve) consistently shows strong binding to the TROP-2 expressing cells, as expected. The various antibody clones show differing levels of binding, with some clones (e.g., trop2-12, trop2-29, and trop2-30) displaying moderate binding strength, while others (e.g., trop2-5 and trop2-27) exhibit weaker binding. This analysis helps to identify the most promising antibody clones for targeting TROP-2 based on their binding affinities.

[0146] The binding activities of isolated TROP-2 specific clones against various cell lines were summarized below. Table 1: Summary of binding activities of clones in FACS.

[0147]

[0148] 293T: TROP-2 high expression by transfection; NCI-H1975 and NCLH929, high expression cancer cell lines; PC3, low expression cancer cell line. Selected Y and N, Y indicates potential candidates, and N indicates candidates not pursued further. NA, not analyzed, W indicates weak binding.

[0149] Example 6:

[0150] Selected clones were further tested in an ELISA, and they displayed various binding affinities as shown in Figure 6.

[0151] This graph shows a dose-response curve for different antibody clones (T3, T7, T8, T12, T21, T24, T29, and T30) based on their binding activity at varying concentrations, measured by ELISA (OD450). The X-axis represents the concentration of the antibody in micrograms per milliliter (pg / mL), while the Y-axis represents the optical density (OD450) values, which indicate the level of binding of the antibodies to the target antigen.

[0152] Binding Strength:

[0153] T8 and T7 show the highest binding activity with a higher OD450 value at lower concentrations, indicating that they bind strongly to the antigen.

[0154] T24 shows the weakest binding, as their curves are lower on the Y-axis, indicating less binding activity.

[0155] 7 clones show an increase in OD450 values as the concentration increases, which is typical in dose-response curves, indicating that more antibodies are available to bind to the antigen. Among the clones, T8, T7, and T12 exhibit stronger and more efficient binding, as shown by their higher OD450 values at comparable concentrations, while T24 shows weaker binding, as indicated by their lower OD450 values even at higher concentrations.

[0156] This graph helps determine which clones have the strongest binding affinity to the target antigen. These insights are valuable for selecting the best candidates for further development in antibody-based applications. The table provides the EC₅₀ values (half maximal effective concentration) for various antibody clones (T3, T7, T8, T12, T21, T24, T29, and T30). The EC₅₀ is a measure of the concentration of the antibody required to achieve 50% of its maximum binding response and the measured EC₅₀ values are shown in the Table 2 below.

[0157] Table 2. EC50 values of different Antibody clones

[0158]

[0159] T8 has the lowest EC₅₀ value (0.04), indicating the strongest binding affinity, as it requires the lowest concentration to achieve half of its maximal binding response.

[0160] T12 also shows strong binding with a low EC₅₀ value (0.0388).

[0161] T30 and T7 have moderate binding affinities, with EC₅₀ values of 0.0621 and 0.2677, respectively.

[0162] T3 has the highest EC₅₀ value (0.597), indicating the weakest binding affinity, as it requires a much higher concentration to reach 50% of its maximal binding.

[0163] T24 is labeled as "NA," which may indicate that the EC₅₀ value could not be determined, potentially due to very weak or no binding.

[0164] Clones T8, T12, and T30 demonstrate the strongest binding affinities, as shown by their low EC₅₀ values, making them strong candidates for further development. T3, with the highest EC₅₀, exhibits the weakest binding, suggesting it may not be a suitable candidate for further development.

[0165] Based on the comprehensive analysis of the binding profiles, the following clones have been selected as potential candidates for further development, and their sequences are reported as they were originally isolated from the scFv libraries.

[0166] Table 3: Amino acid sequences of candidate clones in the original scFv format.

[0167]

[0168]

[0169] Table 4: VH and VL amino acid sequences of candidate clones.

[0170]

[0171]

[0172] Example 7

[0173] Another aspect of this invention relates to “complementarity determining region” or “CDR,” the amino acid sequence of which contributes to antigen binding specificity and affinity. The CDRs include CDR1, CDR2, and CDR3 in the heavy chain variable domain (VH) and CDR1, CDR2, and CDR3 in the light chain variable domain (VL). Consequently, the CDRs of the VHs and VLs described herein have been determined according to the standard numbering scheme, as shown in Table 5 below.

[0174] Table 5 CDR sequences and corresponding SEQ ID numbers

[0175]

[0176]

[0177] As used in this specification and claim(s), the words "comprising" (and any form of comprising, such as "comprise" and "comprises"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "includes" and "include") or "containing" (and any form of containing, such as "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed. To the extent a definition of a term set out in a document incorporated herein by reference conflicts with the definition of a term explicitly defined herein, the definition set out herein controls.

Claims

What is claimed is:

1. An antigen binding protein molecule that specifically binds to human trophoblast cell surface antigen 2 ( TR0P2), wherein the antigen binding protein molecule is selected from the group consisting of an antibody and an antigen binding fragment.

2. The antigen binding protein molecule of claim 1, wherein the antigen binding fragment is selected from the group consisting of a single-chain variable fragment (scFv), a Fab' (fragment antigen-binding, monomeric), a F(ab')2 (Fragment antigen-binding, dimeric), a single chain diabody, and a nanobody.

3. The antigen binding protein molecule of claim 2, wherein the antigen binding fragment is an scFv.

4. The antigen binding protein molecule of claim 3, wherein the scFv is selected from the group consisting of:an scFv comprising VH having the amino acid of SEQ ID NO: 1 and VL having the amino acid of SEQ ID NO: 2;an scFv comprising VH having the amino acid of SEQ ID NO: 3 and VL having the amino acid of SEQ ID NO: 4;an scFv comprising VH having the amino acid of SEQ ID NO: 5 and VL having the amino acid of SEQ ID NO: 6;an scFv comprising VH having the amino acid of SEQ ID NO: 7 and VL having the amino acid of SEQ ID NO: 8;an scFv comprising VH having the amino acid of SEQ ID NO: 9 and VL having the amino acid of SEQ ID NO: 10;an scFv comprising VH having the amino acid of SEQ ID NO: 11 and VL having the amino acid of SEQ ID NO: 12;an scFv comprising VH having the amino acid of SEQ ID NO: 13 and VL having the amino acid of SEQ ID NO: 14;an scFv comprising VH having the amino acid of SEQ ID NO: 15 and VL having the amino acid of SEQ ID NO: 16; andan scFv comprising VH having the amino acid of SEQ ID NO: 17 and VL having the amino acid of SEQ ID NO: 18.

5. The antigen binding protein molecule of claim 3, wherein the scFv is selected from the group consisting ofan scFv comprising the amino acid sequence of SEQ ID NO: 19; an scFv comprising the amino acid sequence of SEQ ID NO: 20;an scFv comprising the amino acid sequence of SEQ ID NO: 21;an scFv comprising the amino acid sequence of SEQ ID NO: 22;an scFv comprising the amino acid sequence of SEQ ID NO: 23;an scFv comprising the amino acid sequence of SEQ ID NO: 24;an scFv comprising the amino acid sequence of SEQ ID NO: 25;an scFv comprising the amino acid sequence of SEQ ID NO: 26; andan scFv comprising the amino acid sequence of SEQ ID NO: 27.

6. The antigen binding protein molecule of claim 3, wherein the scFv is selected from the group consisting of:an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 28, 29, and 30, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 34, 35, and 36, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 40, 41, and 42, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 46, 47, and 48, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 58, 59, and 60, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 64, 65, and 66, respectively;an scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 70, 71, and 72, respectively; andan scFv comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 76, 77, and 78, respectively.

7. The antigen binding protein molecule of claim 3, wherein the scFv is selected from the group consisting ofan scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 31, 32, and 33, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 37, 38, and 39, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 43, 44, and 45, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 49, 50, and 51, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 55, 56, and 57, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 61, 62, and 63, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 67, 68, and 69, respectively;an scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively; andan scFv comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 79, 80, and 81, respectively.

8. The antigen binding protein molecule of claim 1, wherein the antigen binding protein molecule is an antibody.

9. The antigen binding protein molecule of claim 8, wherein the antibody is selected from the group consisting of:an antibody comprising VH having the amino acid of SEQ ID NO: 1 and VL having the amino acid of SEQ ID NO: 2;an antibody comprising VH having the amino acid of SEQ ID NO: 3 and VL having the amino acid of SEQ ID NO: 4;an antibody comprising VH having the amino acid of SEQ ID NO: 5 and VL having the amino acid of SEQ ID NO: 6;an antibody comprising VH having the amino acid of SEQ ID NO: 7 and VL having the amino acid of SEQ ID NO: 8;an antibody comprising VH having the amino acid of SEQ ID NO: 9 and VL having the amino acid of SEQ ID NO: 10;an antibody comprising VH having the amino acid of SEQ ID NO: 11 and VL having the amino acid of SEQ ID NO: 12;an antibody comprising VH having the amino acid of SEQ ID NO: 13 and VL having the amino acid of SEQ ID NO: 14;an antibody comprising VH having the amino acid of SEQ ID NO: 15 and VL having the amino acid of SEQ ID NO: 16; andan antibody comprising VH having the amino acid of SEQ ID NO: 17 and VL having the amino acid of SEQ ID NO: 18.

10. The antigen binding protein molecule of claim 8, wherein the antibody is selected from the group consisting of:an antibody comprising VH having the CDR1, CDR2, and CDR.3 sequences of SEQ ID NOs: 28, 29, and 30, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 34, 35, and 36, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 40, 41, and 42, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 46, 47, and 48, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 58, 59, and 60, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 64, 65, and 66, respectively;an antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 70, 71, and 72, respectively; andan antibody comprising VH having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 76, 77, and 78, respectively.

11. The antigen binding protein molecule of claim 8, wherein the antibody is selected from the group consisting of:an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 31, 32, and 33, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 37, 38, and 39, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 43, 44, and 45, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 49, 50, and 51, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 55, 56, and 57, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 61, 62, and 63, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 67, 68, and 69, respectively;an antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively; andan antibody comprising VL having the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 79, 80, and 81, respectively.

12. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 28,a VH CDR2 having the amino acid of SEQ ID NO: 29,a VH CDR3 having the amino acid of SEQ ID NO: 30,a VL CDR1 having the amino acid of SEQ ID NO: 31,a VL CDR2 having the amino acid of SEQ ID NO: 32, anda VL CDR3 having the amino acid of SEQ ID NO: 33.

13. The antigen binding protein molecule of claim 12, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 1.

14. The antigen binding protein molecule of claim 12, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 2.

15. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 34,a VH CDR2 having the amino acid of SEQ ID NO: 35,a VH CDR3 having the amino acid of SEQ ID NO: 36,a VL CDR1 having the amino acid of SEQ ID NO: 37,a VL CDR2 having the amino acid of SEQ ID NO: 38, anda VL CDR3 having the amino acid of SEQ ID NO: 39.

16. The antigen binding protein molecule of claim 15, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 3.

17. The antigen binding protein molecule of claim 15, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 4.

18. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 40,a VH CDR2 having the amino acid of SEQ ID NO: 41,a VH CDR3 having the amino acid of SEQ ID NO: 42,a VL CDR1 having the amino acid of SEQ ID NO: 43,a VL CDR2 having the amino acid of SEQ ID NO: 44, anda VL CDR3 having the amino acid of SEQ ID NO: 45.

19. The antigen binding protein molecule of claim 18, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 5.

20. The antigen binding protein molecule of claim 18, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 6.

21. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 46,a VH CDR2 having the amino acid of SEQ ID NO: 47,a VH CDR3 having the amino acid of SEQ ID NO: 48,a VL CDR1 having the amino acid of SEQ ID NO: 49,a VL CDR2 having the amino acid of SEQ ID NO: 50, anda VL CDR3 having the amino acid of SEQ ID NO: 51.

22. The antigen binding protein molecule of claim 21, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 7.

23. The antigen binding protein molecule of claim 21, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 8.

24. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 52,a VH CDR2 having the amino acid of SEQ ID NO: 53,a VH CDR3 having the amino acid of SEQ ID NO: 54,a VL CDR1 having the amino acid of SEQ ID NO: 55,a VL CDR2 having the amino acid of SEQ ID NO: 56, anda VL CDR3 having the amino acid of SEQ ID NO: 57.

25. The antigen binding protein molecule of claim 24, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 9.

26. The antigen binding protein molecule of claim 24, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 10.

27. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 58,a VH CDR2 having the amino acid of SEQ ID NO: 59,a VH CDR3 having the amino acid of SEQ ID NO: 60,a VL CDR1 having the amino acid of SEQ ID NO: 61,a VL CDR2 having the amino acid of SEQ ID NO: 62, anda VL CDR3 having the amino acid of SEQ ID NO: 63.

28. The antigen binding protein molecule of claim 27, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 11.

29. The antigen binding protein molecule of claim 27, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 12.

30. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 64,a VH CDR2 having the amino acid of SEQ ID NO: 65,a VH CDR3 having the amino acid of SEQ ID NO: 66,a VL CDR1 having the amino acid of SEQ ID NO: 67,a VL CDR2 having the amino acid of SEQ ID NO: 68, anda VL CDR3 having the amino acid of SEQ ID NO: 69.

31. The antigen binding protein molecule of claim 30, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 13.

32. The antigen binding protein molecule of claim 27, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 14.

33. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 70,a VH CDR2 having the amino acid of SEQ ID NO: 71,a VH CDR3 having the amino acid of SEQ ID NO: 72,a VL CDR1 having the amino acid of SEQ ID NO: 73,a VL CDR2 having the amino acid of SEQ ID NO: 74, anda VL CDR3 having the amino acid of SEQ ID NO: 75.

34. The antigen binding protein molecule of claim 33, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 15.

35. The antigen binding protein molecule of claim 33, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 16.

36. An antigen binding protein molecule, being an antibody or an scFv, comprising:a VH CDR1 having the amino acid of SEQ ID NO: 76,a VH CDR2 having the amino acid of SEQ ID NO: 77,a VH CDR3 having the amino acid of SEQ ID NO: 78,a VL CDR1 having the amino acid of SEQ ID NO: 79,a VL CDR2 having the amino acid of SEQ ID NO: 80, anda VL CDR3 having the amino acid of SEQ ID NO: 81.

37. The antigen binding protein molecule of claim 36, wherein the antigen binding protein molecule comprises a VH amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 17.

38. The antigen binding protein molecule of claim 36, wherein the antigen binding protein molecule comprises a VL amino acid sequence that is at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid of SEQ ID NO: 18.

39. The antigen binding protein molecule of one of claims 12-38, wherein the antigen binding protein molecule is an antibody, comprising a variable region and a constant region, capable of specifically binding to human TROP2.

40. The antigen binding protein molecule of claim 39, wherein the antibodby is a humanized antibody.

41. The antigen binding protein molecule of one of claims 12-38, wherein the antigen binding protein molecule is an scFv, comprising the variable regions of the heavy and light chains connected by a linker peptide, and capable of specifically binding to human TROP2.

42. A pharmaceutical composition comprising the antigen binding protein molecule of one of claims 4-38, wherein the antigen binding protein molecule is formulated for therapeutic use in treating a disease associated with human TROP2 overexpression.

43. A nucleic acid encoding the antigen binding protein molecule of one of claims 4-38, wherein the nucleic acid sequence encodes the variable regions responsible for binding to human TROP2, and is suitable for expression in a host cell for therapeutic or diagnostic purposes.

44. A vector comprising the nucleic acid of claim 43.

45. A cell comprising the vectors of claim 44, wherein the host cell is selected from a mammalian cell, bacterial cell, yeast cell, or insect cell, and is capable of expressing the antigen binding protein molecule encoded by the nucleic acid molecule.

46. A method of making an antigen binding molecule of claims 4-38 comprising culturing the cell of claim 45 under suitable conditions.

47. A method of a cell-based therapy, comprising administering to a subject in need thereof a preselected number of cells of claim 45 expressing a therapeutic dose of the antigen binding protein molecule of claim 39 or claim 41.

48. A method of administering a dose of a medicament, comprising administering to a subject in need thereof an effective dose of a composition, wherein the composition comprises the pharmaceutical composition of claim 42.

49. A method of treating a disease, the method comprising administering to a subject in need thereof an effective amount of a composition, wherein the composition comprises the pharmaceutical composition of claim 42, wherein the administration is carried out by a delivery method selected from the group consisting of intravenous, subcutaneous, or intraperitoneal injection.

50. A method of treating cancer in a human subject, the method comprising administering to the subject an effective amount of a composition, wherein the composition comprises the pharmaceutical composition of claim 42 and the cancer is selected from the group consisting of epithelial cancers, breast cancer, lung cancer, ovarian cancer, colorectal cancer, pancreatic cancer, and prostate cancer, wherein the composition is administered via intravenous or subcutaneous injection.

51. The method of claim 49, wherein the subject is a human.

52. A method of inhibiting or suppressing a tumor in a subject, comprising administering an effective amount of a composition to the subject, wherein the composition comprises the pharmaceutical composition of claim 42.

53. A method of delaying progression of a solid tumor in a subject, comprising administering an effective amount of a composition to the subject, wherein the composition comprises the pharmaceutical composition of claim 42, and wherein the solid tumor is selected from the group consisting of epithelial cancers, breast cancer, lung cancer, ovarian cancer, colorectal cancer, pancreatic cancer, and gastric cancer, and the administration is carried out via intravenous or subcutaneous injection..

54. The antigen binding protein molecule of claim 39 or claim 41, further comprises a detectable label.

55. The antigen binding protein molecule of claim 54, wherein the detectable label is selected from the group consisting of a fluorescent label, a bioluminescent protein / peptide, a photochromic compound, a radiolabel, and a hapten.

56. A method of detecting the presence of human TROP2 in a sample, comprising contacting the sample with a composition comprising the antigen binding protein molecule of claim 54, whereby a positive measurement of the detectable label indicates the presence of human TROP2 in the sample, wherein the biological sample is selected from the group consisting of blood, serum, tissue biopsy, or cell lysate.

57. A method of quantifying the amount of human TROP2 in a sample, comprising contacting the sample with a composition comprising the antigen binding protein molecule of claim 54,whereby a numeric measurement of the detectable label indicates the amount of human TROP2 in the sample.

58. A method of diagnosing the presence of a tumor or a cancer growth through a clinical sample, comprising:(a) obtaining a clinical sample from a patient in need thereof, wherein the clinical sample is selected from blood, serum, tissue biopsy, or cell lysate; and,(b) contacting the clinical sample with the antigen binding protein molecule of claim 54; whereby a positive measurement of the detectable label indicates the presence of a tumor or cancer growth associated with TR0P2 expression in the patient.

59. A method of diagnosing the presence of a tumor or a cancer growth in a subject through in vivo imaging, comprising:(a) administering to the subject a diagnostically effective amount of a composition comprising the antigen binding protein molecule of claim 54, wherein the detectable label is suitable for the in vivo imaging; and(b) applying whole-body or regionalized imaging to detect the bound antigen binding protein molecule by the detectable label;whereby a positive measurement of the detectable label indicates the presence of a tumor or cancer growth associated with TROP2 expression in the subject.

60. A kit of detecting human TROP2, comprising the antigen binding protein molecule of claim 54.

61. A kit of quantifying the amount of human TROP2 in a sample, comprising the antigen binding protein molecule of claim 54.

Images