Degradation of cMET using bispecific binding agents

Abstract

The present disclosure provides methods of degrading a cMET protein on a target cell. The present disclosure further discloses binding agents that bind to the cMET protein and the degradation protein.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 384,371, filed on November 18, 2022, and U.S. Provisional Patent Application No. 63 / 479,515, filed on January 11, 2023, each of which is incorporated herein by reference in its entirety. Background Art

[0003] Compared to traditional inhibition-based therapies, targeted protein degradation is a promising new therapeutic strategy. Inhibitors rely on sustained, occupancy-driven pharmacology, requiring high-affinity binders that can eliminate catalytic or binding functions. For standard binding-based small molecules, inhibiting protein-protein interactions or scaffold functions has been extremely challenging. In contrast, protein degraders are catalytic and utilize event-driven pharmacology, alleviating the need for high-affinity binders and persistently eliminating all protein functions at once. As a result, degrader technologies such as protein degradation targeting chimeras (PROTACs) have achieved great success in targeting traditionally challenging proteins. Many PROTACs are currently undergoing clinical trials.

[0004] Most degradation agent technologies (including PROTAC) utilize intracellular mechanisms of action and are therefore largely limited to targeting proteins with cytoplasmic domains. However, methods for specific degradation of cell surface proteins, such as LYTAC, have recently emerged. These methods utilize recycled glycan receptors (such as mannose-6-phosphate receptor (M6PR) or asialoglycoprotein receptor (ASGR) etc.) to target proteins for internalization and transport to lysosomes for degradation. These methods require complex glycans conjugated to antibodies or small molecules to achieve degradation of membrane proteins.

[0005] As a hybrid approach that is broadly applicable to many cell types, we recently described antibody-based PROTACs (AbTACs). AbTACs utilize a standard IgG bispecific antibody format to bring a cell surface E3 ligase (RNF43) close to a membrane protein of interest (POI) to mediate its degradation via the lysosomal pathway. The traditional bispecific IgG scaffold on which AbTACs are constructed has good pharmacokinetic properties relative to LYTACs and other small molecule-based degraders. In addition, AbTACs are fully recombinant compared to other degradation modalities (such as LYTACs and PROTACs). However, there is still a need for targeted protein degraders that efficiently and selectively induce degradation of target proteins. Summary of the Invention

[0006] In one aspect, the present disclosure provides a method for degrading a target protein on the surface of a target cell, the method comprising: contacting a degradation protein and the target protein on the surface of the target cell with a binding agent, wherein the binding agent comprises: (i) a first binding domain, which specifically binds to the degradation protein; (ii) a second binding domain, which specifically binds to the target protein, wherein the target protein comprises cMET.

[0007] In some embodiments, the binding agent is a multispecific antibody, a bispecific antibody, a bispecific diabody, a bispecific Fab2, a bispecific camelid antibody, a bispecific peptibody scFv-Fc, a bispecific IgG, a knob and hole bispecific IgG, an Fc-Fab, or a knob and hole bispecific Fc-Fab. In some embodiments, the binding agent is a multispecific antibody or a bispecific antibody. In some embodiments, the binding agent is a bispecific antibody.

[0008] In some embodiments, the degraded protein is CDH3, MUC1, CD276, TROP2, CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, EGFR, MST1R, EphA2, ADAM9, IGF1R, RNF43, RNF128, RNF130, or ZNRF3.

[0009] In some embodiments, the degradation protein is CDH3. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the variable heavy chain of the first binding domain comprises at least 80% sequence identity with any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

[0010] In some embodiments, the degradation protein is MUC1. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 6, 10, 14, 18, or 22 and any of SEQ ID NOs: 8, 12, 16, 20, or 24.

[0011] In some embodiments, the degradation protein is CD276. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 25, 26, 30, 34, or 38. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope e of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 26, 30, 34, or 38 and any of SEQ ID NOs: 28, 32, 36, or 40.

[0012] In some embodiments, the degradation protein is TROP2. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

[0013] In some embodiments, the degradation protein is selected from CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, MST1R, EphA2, ADAM9, IGF1R and EGFR. In some embodiments, the degradation protein is RNF43, RNF128, RNF130 or ZNRF3. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of the variable heavy chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of the variable heavy chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable heavy chain comprises any one of the variable heavy chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable light chain comprises any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4.

[0014] In some embodiments, the second binding domain comprises a second binding domain variable heavy chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain variable heavy chain comprises any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain comprises a second binding domain variable light chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366. In some embodiments, the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366. In some embodiments, the second binding domain variable heavy chain comprises any one of SEQ ID NO: 354, 358, 362, or 366.

[0015] In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by Amivantamab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by Amivantamab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope does not comprise any one of the amino acids from the epitope bound by Amivantamab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by Telisotuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by Telisotuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, and the epitope does not include any one of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by Onartuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by Onartuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by Onartuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by REGN5093s58.

[0016] In some embodiments, after the contacting, cMET is internalized into the target cell along with the degradation protein, and cMET is degraded. In some embodiments, after the cMET is internalized into the target cell along with the degradation protein, the degradation protein is recycled to the surface of the target cell. In some embodiments, after the cMET is internalized into the target cell along with the degradation protein, one or more of the degradation protein or cMET is degraded.

[0017] In some embodiments, the target cell is a cancer cell. In some embodiments, the cancer cell is selected from breast cancer cells, B cell lymphoma cells, pancreatic cancer cells, Hodgkin lymphoma cells, ovarian cancer cells, prostate cancer cells, mesothelioma cells, lung cancer cells, non-Hodgkin B cells (B-NHL) cells, melanoma cells, chronic lymphocytic leukemia cells, acute lymphocytic leukemia cells, neuroblastoma cells, glioma cells, glioblastoma cells, bladder cancer cells, colorectal cancer cells, gastric adenocarcinoma cells, non-small cell lung cancer cells, head and neck cancer cells, and cancers carrying cMET mutations including exon 14 deletions. In some embodiments, the cancer cell is a gastric adenocarcinoma cell. In some embodiments, the cancer cell is a non-small cell lung cancer cell. In some embodiments, the cancer cell comprises a gene mutation selected from a cMET exon 14 skipping mutation or a cMET duplication mutation. In some embodiments, the mutation comprises a cMET exon 14 skipping mutation. In some embodiments, the cancer cell comprises a cMET duplication mutation.

[0018] In some embodiments, the expression of cMET in the cancer cell after said contact with said binding agent is less than the expression of cMET in a control cancer cell not contacted with said binding agent. In some embodiments, the expression of cMET in the cancer cell after said contact with said binding agent is at least 50% less than the expression of cMET in a control cancer cell not contacted with said binding agent. In some embodiments, the expression of cMET in the cancer cell after said contact with said binding agent is at least 50% less than the expression of cMET in a control cancer cell contacted with a monospecific cMET binding agent.

[0019] In some embodiments, the amount of cMET on the surface of the cancer cell after contacting with the binding agent is less than the amount of cMET on the surface of a control cancer cell that has not been contacted with the binding agent. In some embodiments, the amount of cMET on the surface of the cancer cell after contacting with the binding agent is at least 20% less than the amount of cMET on the surface of a control cancer cell that has not been contacted with the binding agent. In some embodiments, the amount of cMET on the surface of the cancer cell after contacting with the binding agent is at least 20% less than the amount of cMET on the surface of a control cancer cell that has not been contacted with the binding agent. In some embodiments, the amount of cMET on the surface of the cancer cell after contacting with the binding agent is at least 20% less than the amount of cMET on the surface of a control cancer cell that has been contacted with a monospecific cMET binding agent. In some embodiments, the amount of cMET on the surface of the cancer cell is determined by staining the cancer cell with a fluorescently labeled antibody to cMET and measuring the fluorescence intensity.

[0020] In some embodiments, the amount of cMET internalized by the cancer cell after said contact with the binding agent is greater than the amount of cMET internalized by a control cancer cell that has not been contacted with the binding agent. In some embodiments, the amount of cMET internalized by the cancer cell after said contact with the binding agent is at least 20% greater than the amount of cMET internalized by a control cancer cell that has not been contacted with the binding agent. In some embodiments, the amount of cMET internalized by the cancer cell after said contact with the binding agent is at least 20% greater than the amount of cMET internalized by a control cancer cell that has not been contacted with the binding agent. In some embodiments, the amount of cMET internalized by the cancer cell after said contact with the binding agent is at least 20% greater than the amount of cMET internalized by a control cancer cell that has been contacted with a monospecific cMET binding agent. In some embodiments, the amount of internalized cMET is determined by the further steps of labeling the binding agent with a fluorescent tag prior to said contact with the degradation protein and the target protein and measuring the fluorescence of the fluorescent tag after contact with the degradation protein and the target protein, wherein the fluorescent tag selectively fluoresces at intracellular pH.

[0021] In some embodiments, the amount of cMET degraded in the cancer cell after said contact with the binding agent is greater than the amount of cMET degraded in a control cancer cell not contacted with the binding agent. In some embodiments, the amount of cMET degraded in the cancer cell after said contact with the binding agent is at least 20% greater than the amount of cMET degraded in a control cancer cell not contacted with the binding agent. In some embodiments, the amount of cMET degraded in the cancer cell after said contact with the binding agent is at least 20% greater than the amount of cMET degraded in a control cancer cell contacted with a monospecific cMET binding agent.

[0022] In some embodiments, the amount of cMET dimer on the cancer cell after said contact with the binding agent is less than the amount of cMET dimer on a control cancer cell not contacted with the binding agent. In some embodiments, the amount of cMET dimer on the cancer cell after said contact with the binding agent is less than the amount of cMET dimer on a control cancer cell contacted with a monospecific cMET binding agent. In some embodiments, the amount of cMET activation in the cancer cell after said contact with the binding agent is within 50% of the amount of cMET activation in a control cancer cell not contacted with the binding agent.

[0023] In some embodiments, the monospecific cMET-binding agent is terituzumab. In some embodiments, the monospecific cMET-binding agent is onatuzumab. In some embodiments, the monospecific cMET-binding agent is REGN5093s58.

[0024] In some embodiments, the method increases the sensitivity of the cancer cells to a cancer therapeutic agent or radiation therapy. In some embodiments, the cancer therapeutic agent is a cytotoxic agent. In some embodiments, the method reduces the proliferation of the cancer cells. In some embodiments, the method induces the death of the cancer cells. In some embodiments, the contacting is performed in vivo.

[0025] In one aspect, the present disclosure provides a method of treating cancer in a subject in need thereof, the method comprising: administering a binding agent to the subject, wherein the binding agent comprises: (i) a first binding domain that specifically binds to a degradation protein, wherein the degradation protein is expressed on a target cell; and (ii) a second binding domain that specifically binds to the target protein, wherein the target protein comprises cMET.

[0026] In some embodiments, the binding agent is a multispecific antibody, a bispecific antibody, a bispecific diabody, a bispecific Fab2, a bispecific camelid antibody, a bispecific peptibody scFv-Fc, a bispecific IgG, a knob-in-hole bispecific IgG, an Fc-Fab, or a knob-in-hole bispecific Fc-Fab. In some embodiments, the binding agent is a multispecific antibody or a bispecific antibody. In some embodiments, the binding agent is a bispecific antibody.

[0027] In some embodiments, the degraded protein is CDH3, MUC1, CD276, TROP2, CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, EGFR, MST1R, EphA2, ADAM9, IGF1R, RNF43, RNF128, RNF130, or ZNRF3.

[0028] In some embodiments, the degradation protein is CDH3. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the variable heavy chain of the first binding domain comprises at least 80% sequence identity with any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

[0029] In some embodiments, wherein the degradation protein is MUC1. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 6, 10, 14, 18, or 22 and any of SEQ ID NOs: 8, 12, 16, 20, or 24.

[0030] In some embodiments, the degradation protein is CD276. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope e of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 26, 30, 34, or 38 and any of SEQ ID NOs: 28, 32, 36, or 40.

[0031] In some embodiments, the degradation protein is TROP2. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

[0032] In some embodiments, the degradation protein is selected from CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, MST1R, EphA2, ADAM9, IGF1R and EGFR. In some embodiments, the degradation protein is RNF43, RNF128, RNF130 or ZNRF3. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of the variable heavy chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of the variable heavy chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable heavy chain comprises any one of the variable heavy chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain variable light chain comprises any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4.

[0033] In some embodiments, the second binding domain comprises a second binding domain variable heavy chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain variable heavy chain comprises any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain comprises a second binding domain variable light chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366. In some embodiments, the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366. In some embodiments, the second binding domain variable heavy chain comprises any one of SEQ ID NO: 354, 358, 362, or 366.

[0034] In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope does not comprise any of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, and the epitope does not include any of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by onatuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by onatuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by onatuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by REGN5093s58.

[0035] In some embodiments, the cancer cell is selected from breast cancer cells, B cell lymphoma cells, pancreatic cancer cells, Hodgkin lymphoma cells, ovarian cancer cells, prostate cancer cells, mesothelioma cells, lung cancer cells, non-Hodgkin B cells (B-NHL) cells, melanoma cells, chronic lymphocytic leukemia cells, acute lymphocytic leukemia cells, neuroblastoma cells, glioma cells, glioblastoma cells, bladder cancer cells, colorectal cancer cells, gastric adenocarcinoma cells, non-small cell lung cancer cells, head and neck cancer cells, and cancers carrying cMET mutations including exon 14 deletions. In some embodiments, the cancer cell is a gastric adenocarcinoma cell. In some embodiments, the cancer cell is a non-small cell lung cancer cell. In some embodiments, the cancer cell comprises a genetic mutation selected from a cMET exon 14 skipping mutation or a cMET duplication mutation. In some embodiments, the mutation comprises a cMET exon 14 skipping mutation. In some embodiments, the cancer cell comprises a cMET duplication mutation.

[0036] In some embodiments, the methods increase the sensitivity of cancer cells to a cancer therapeutic agent or radiation therapy. In some embodiments, the cancer therapeutic agent is a cytotoxic agent. In some embodiments, the methods reduce the proliferation of cancer cells. In some embodiments, the methods induce the death of cancer cells.

[0037] In one aspect, the present disclosure provides a binding agent comprising: (a) a first binding domain that specifically binds to a degradation protein, wherein the degradation protein is CDH3, MUC1, CD276 or TROP2; and (b) a second binding domain that specifically binds to a target protein, wherein the target protein is cMET.

[0038] In some embodiments, the multispecific binding agent is a multispecific antibody, a bispecific antibody, a bispecific diabody, a bispecific Fab2, a bispecific camelid antibody, a bispecific peptibody scFv-Fc, a bispecific IgG, a knob-in-hole bispecific IgG, an Fc-Fab, or a knob-in-hole bispecific Fc-Fab. In some embodiments, the binding agent is a multispecific antibody or a bispecific antibody. In some embodiments, the binding agent is a bispecific antibody.

[0039] In some embodiments, the degradation protein is CDH3. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the variable heavy chain of the first binding domain comprises at least 80% sequence identity with any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 106, 110, 114, 118, or 122. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

[0040] In some embodiments, the degradation protein is MUC1. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 6, 10, 14, 18, or 22. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 6, 10, 14, 18, or 22 and any of SEQ ID NOs: 8, 12, 16, 20, or 24.

[0041] In some embodiments, the degradation protein is CD276. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 26, 30, 34, or 38. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity with any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity with any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope e of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not include any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 26, 30, 34, or 38 and any of SEQ ID NOs: 28, 32, 36, or 40. In some embodiments, the degraded protein is TROP2. In some embodiments, the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any of SEQ ID NOs: 198, 202, 206, 210, or 214.In some embodiments, the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain variable heavy chain comprises any one of SEQ ID NOs: 198, 202, 206, 210, or 214. In some embodiments, the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain variable light chain comprises any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216. In some embodiments, the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of SEQ ID NOs: 198, 202, 206, 210, or 214 and any of SEQ ID NOs: 200, 204, 208, 212, or 216.

[0042] In some embodiments, the second binding domain comprises a second binding domain variable heavy chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain variable heavy chain comprises any one of SEQ ID NOs: 352, 356, 360, or 364. In some embodiments, the second binding domain comprises a second binding domain variable light chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366. In some embodiments, the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366. In some embodiments, the second binding domain variable heavy chain comprises any one of SEQ ID NO: 354, 358, 362, or 366.

[0043] In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope does not comprise any of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, and the epitope does not include any of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by onatuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by onatuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity with the epitope bound by onatuzumab. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by REGN5093s58.

[0044] Incorporation by reference

[0045] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent that publications and patents or patent applications incorporated by reference contradict the disclosure contained in this specification, the specification is intended to supersede and / or take precedence over any such contradictory material. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The novel features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by referring to the following detailed description and accompanying drawings (also referred to herein as "figures" and "FIG.") which illustrate illustrative embodiments utilizing the principles of the present disclosure, wherein:

[0047] Figure 1 The methods of the present disclosure are depicted, wherein degradation of a target protein 112 (ie, cMET) is mediated by binding of a bifunctional binding agent 101 .

[0048] Figure 2A-2D is a graph depicting the percentage of cMET cell surface removal in various cell types when treated with bispecific antibodies. Figure 2A is a graph depicting the percentage of cMET cell surface depletion in Hs746T cells treated with bispecific antibodies. Figure 2B is a graph depicting the percentage of cMET cell surface depletion in NCI-H1993 cells treated with bispecific antibodies. Figure 2C is a graph depicting the percentage of cMET cell surface depletion in NCI-H1975 cells treated with bispecific antibodies. Figure 2D is a graph depicting the percentage of cMET cell surface depletion in NCI-H596 cells treated with bispecific antibodies.

[0049] Figure 3A-Figure 3B is a graph depicting the percentage of cell surface depletion of cMET on target cells treated with various bispecific antibodies. Figure 3A is a graph depicting the percentage of cell surface depletion of cMET on NCI-H1975 target cells treated with various bispecific antibodies at a concentration of 50 nM. Figure 3B is a graph depicting the percentage of cell surface depletion of cMET on NCI-H1975 target cells treated with various bispecific antibodies at a concentration of 50 nM.

[0050] Figures 4A-4C is a graph depicting cell surface removal of cMET. Figure 4Ais a graph depicting cell surface removal of cMET on NCI-H1975 target cells when treated with various bispecific antibodies, wherein the antibodies to the cMET target bind to different epitopes. Figure 4B is a graph depicting cell surface removal of cMET on NCI-H596 target cells when treated with various bispecific antibodies, where the antibodies to the degraders bind to different epitopes. Figure 4C is a graph depicting cell surface removal of cMET on Hs746T target cells when treated with various bispecific antibodies, where the antibodies to the degraders bind to different epitopes.

[0051] Figure 5 is a graph depicting internalization of cMET on target cells when treated with various bispecific antibodies, where the bispecific drove higher internalization than either single-armed mAb targeting either the target or the degrader.

[0052] Figures 6A-6C is a graph depicting the degradation of cMET on target cells when treated with various bispecific antibodies. Figure 6A is a graph depicting the internalization of cMET on NCI-H1975 target cells when treated with various bispecific antibodies. Figure 6B is a graph depicting whole cell degradation on HS746t target cells when treated with various bispecific antibodies. Figure 6C is a graph depicting whole cell degradation on NCI-H596 target cells when treated with various bispecific antibodies.

[0053] Figures 7A-7C Depicted are the amounts of cMET on target cells treated with various bispecific antibodies. Figure 7A are images of Western blots depicting the amount of cMET protein on target cells when treated with various bispecific antibodies. Figure 7B are images of Western blots depicting the amount of cMET protein on target cells when treated with various bispecific antibodies. Figure 7C is a graph depicting whole-cell degradation of cMET on target cells when treated with various bispecific antibodies.

[0054] Figures 8A-8E Depicted are the amounts of pERK and ERK in target cells treated with various bispecific antibodies. Figure 8A are images of Western blots depicting the amounts of pERK and ERK proteins in target cells when treated with various bispecific antibodies at different concentrations. Figure 8B are images of Western blots depicting the amounts of pERK and ERK proteins in target cells when treated with various bispecific antibodies at different concentrations. Figure 8C is a graph depicting the percentage of pERK to ERK in target cells when treated with various bispecific antibodies at different concentrations. Figure 8D is a graph depicting the percentage reduction in the amount of cMET and the ratio of pERK to ERK in target cells when treated with various bispecific antibodies at different concentrations compared to PBS. Figure 8E is a graph depicting the percentage reduction in the amount of cMET and the ratio of pERK to ERK in target cells when treated with various bispecific antibodies at different concentrations.

[0055] Figure 9 are images of Western blots depicting the amount of cMET protein on Hs746t target cells when treated with various bispecific antibodies.

[0056] Figures 10A-10D Depicted is the amount of cMET dimerization on target cells treated with various bispecific antibodies. Figure 10A is a graph showing dimerization of cMET on target cells when treated with hepatocyte growth factor, the natural ligand of cMET. Figure 10B are graphs showing cMET dimerization on target cells when treated with various bispecific antibodies comprising the eviantazumab cMET-binding arm. Figure 10C are graphs showing cMET dimerization on target cells when treated with various bispecific antibodies comprising the onatumomab cMET binding arm. Figure 10D are graphs showing cMET dimerization on target cells when treated with various bispecific antibodies comprising the terituzumab cMET-binding arm. DETAILED DESCRIPTION

[0057] The present disclosure generally relates to binding agents that bind to both a target protein and a degradation protein present on the surface of a target cell. In some embodiments, the present disclosure provides a method for degrading a target protein, comprising contacting the target protein with a dual binding agent and a degradation protein that simultaneously binds, resulting in cellular internalization of the target protein and subsequent degradation of the target protein.

[0058] definition

[0059] Unless defined otherwise, 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 disclosure belongs. All patents and publications cited herein are incorporated by reference.

[0060] As used in this specification and the claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0061] The terms "administer," "administered," "administers," and "administering" are defined as providing a composition to a subject via routes known in the art, including but not limited to intravenous, intraarterial, intrathecal, oral, parenteral, perineural, buccal, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, intraperitoneal, or nerve root sheath administration. In certain embodiments, the composition can be administered using an oral route. The terms "administer," "administered," "administers," and "administering" a therapeutic protein should be understood to mean providing a therapeutic protein of the disclosure or a prodrug of a therapeutic protein of the disclosure to an individual in need thereof.

[0062] The term "humanization" refers to the replacement or substitution of certain amino acids in antibodies or Nanobodies derived from non-human species, in particular in the framework regions and constant domains of the heavy and / or light chains, in order to avoid or minimize an immune response in humans.

[0063] As used herein, the term "complementarity determining region" or "CDR" in the context of an antibody or nanobody refers to the variable region of H (heavy chain) or L (light chain) (also abbreviated as VH and VL, respectively), and contains an amino acid sequence that can specifically bind to an antigen target. These CDR regions explain the basic specificity of an antibody to a particular antigenic determinant structure. Such regions are also referred to as "hypervariable regions". CDRs represent discontinuous segments of amino acids within the variable region, but, regardless of species, the positional positions of these key amino acid sequences within the variable heavy and light chain regions have been found to be similar within the amino acid sequence of the variable chain. The variable heavy and light chains of all typical antibodies each have three CDR regions, and each CDR region of the corresponding light (L) and heavy (H) chains is discontinuous with the other CDR regions (referred to as L1, L2, L3, H1, H2, H3). In particular, nanobodies typically comprise a single amino acid chain, which can be considered to comprise four "framework sequences or framework regions" (or FRs) and three "complementarity determining regions" (or CDRs). Nanobodies have three CDR regions, each of which is discontinuous with the other CDR regions (referred to as CDR1, CDR2, and CDR3). The description of the FR and CDR sequences is based on the IMGT unique numbering system for V domains and V-like domains.

[0064] As used herein, the terms "nucleic acid molecule," "polynucleotide," "polynucleic acid," and "nucleic acid" are used interchangeably and refer to a polymeric form of nucleotides of any length (deoxyribonucleotides or ribonucleotides, or analogs thereof). A polynucleotide can have any three-dimensional structure and can perform any function, known or unknown. Non-limiting examples of polynucleotides include genes, gene fragments, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, control regions, isolated RNA of any sequence, nucleic acid probes, and primers. Nucleic acid molecules can be linear or circular.

[0065] As used herein, "nanoantibodies" (Nb) refer to the smallest antigen-binding fragments or single variable domains ("VHH") derived from naturally occurring heavy chain antibodies and are known to those skilled in the art. They are derived from heavy chain antibodies only, as found in, for example, camel antibodies. Nanoantibodies herein generally comprise a single amino acid chain, which can be considered to comprise four "framework sequences" and three "complementarity determining regions" (or CDRs, as defined above) that constitute a "scaffold". It should be noted that the term "nanoantibody", as used herein in its broadest sense, is not limited to a specific biological source or a specific method of preparation.

[0066] The phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0067] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0068] As used herein, the terms "polypeptide," "protein," and "peptide" are used interchangeably herein and refer to a polymeric form of amino acids of any length, which may include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides with modified peptide backbones.

[0069] The terms "subject," "individual," and "patient" are used interchangeably and refer to humans as well as non-human mammals (e.g., non-human primates, dogs, horses, cats, pigs, cattle, ungulates, lagomorphs, rodents, etc.). In various embodiments, a subject can be a human (e.g., an adult male, an adult female, an adolescent male, an adolescent female, a male child, a female child) under the care of a physician or other health worker in a hospital, outpatient clinic, or other clinical setting. In certain embodiments, a subject may not be under the care or prescription of a physician or other health worker.

[0070] As used herein, the phrase "a subject in need thereof" refers to a subject having or at risk for a pathology to be treated prophylactically or therapeutically with a therapeutic protein as described herein, as described below.

[0071] As used herein, the term "specificity" refers to the ability of a protein binding domain, particularly an immunoglobulin or immunoglobulin fragment (such as a Nanobody), to preferentially bind one antigen over a different antigen and does not necessarily imply high affinity.

[0072] As used herein, "treatment" or "treating" refers to a method for obtaining a beneficial or desired result (including but not limited to therapeutic benefit and / or preventive benefit) with respect to a disease, disorder or medical condition. In certain embodiments, treatment or treating involves administering a therapeutic protein or composition disclosed herein to a subject. Therapeutic benefit can include eradicating or improving the underlying condition being treated. In addition, therapeutic benefit can be achieved by eradicating or improving one or more physiological symptoms associated with the underlying condition (such as observing an improvement in the subject, although the subject may still suffer from the underlying condition). In certain embodiments, for preventive benefit, the composition is administered to a subject at risk of developing a particular disease, or to a subject reporting one or more physiological symptoms of a disease, even if a diagnosis of the disease may not yet be made. Treatment can include, for example, reducing, delaying or alleviating the severity of one or more symptoms of a disease or condition, or it can include reducing the frequency of symptoms of a disease, defect, disorder or adverse condition experienced by the patient. Treatment can be used herein to refer to a method that results in some degree of treatment or improvement of a disease or condition, and a range of results for this purpose can be expected, including but not limited to preventing the condition completely.

[0073] In certain embodiments, the terms "prevent" or "preventing" in relation to a disease or condition may refer to reducing the occurrence of the disease or condition in a treated sample relative to an untreated control sample, or delaying the onset or reducing the severity of one or more symptoms of the disease or condition relative to an untreated control sample in a statistical sample.

[0074] As used herein, the term "therapeutic effect" encompasses therapeutic benefits and / or prophylactic benefits as described above. A prophylactic effect includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, arresting or reversing the progression of a disease or condition, or any combination thereof.

[0075] As used herein, the term "degradation protein" or "degrader protein" can encompass a range of moieties including, but not limited to, membrane-associated internalization proteins, internalization receptors, membrane-associated degradation receptors, degradation receptors, surface moieties configured as internalization binders, surface moieties configured as degradation binders, combinations thereof, or variants thereof.

[0076] The term "internalization protein" as used herein can encompass a range of moieties including, but not limited to, membrane-associated internalization proteins, internalization receptors, surface moieties configured as internalization binding agents, combinations thereof, or variants thereof.

[0077] Methods for degrading cMET protein

[0078] Mesenchymal epithelial transition factor (cMET) is a transmembrane protein that is a receptor for hepatocyte growth factor / scatter factor ligands. cMET is a receptor tyrosine kinase that is activated by binding to these specific ligands, including hepatocyte growth factor (HGF), and subsequent dimerization. Aberrant cMET function and / or expression is associated with cancer, where it leads to enhanced cell proliferation and drives tumor growth, invasion, metastasis, and angiogenesis.

[0079] Mutations that lead to overexpression (called upregulation or amplification) of cMET are associated with many cancers, including colorectal cancer, non-small cell lung cancer, breast cancer, renal cell carcinoma, and head and neck cancer. High expression of cMET is associated with poor prognosis in cancer patients, and abnormal activation of cMET is associated with resistance to targeted therapies. Upregulation or overactivation of cMET can induce a variety of signaling cascades that lead to motility, invasion, growth, and transformation. Therefore, degradation of cMET in cancer is a promising cancer treatment approach.

[0080] The present disclosure provides a method for degrading cMET protein on target cells, such as Figure 1As shown. The method utilizes a binding agent 101 that specifically binds to both: (i) an extracellular epitope on a cMET protein 112; and (2) an extracellular epitope on a membrane-associated internalization protein 113 on a target cell 111. The binding agent 101 comprises a first binding domain 102 that selectively binds to the cMET protein 112 and a second binding domain 103 that selectively binds to the membrane-associated internalization protein 113. Simultaneous binding of the binding agent 101 to the cMET protein 112 and the membrane-associated internalization protein 113 results in both the cMET protein 112 and the membrane-associated internalization protein 113 being internalized into the target cell 111. After internalization, the cMET protein 112 is degraded by the target cell 111 (e.g., via trafficking to lysosomes).

[0081] In some embodiments, the membrane-associated internalization protein is a cell surface protein that is internalized after a binding agent (e.g., an antibody) binds to the protein. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123 , ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, L Y6E, CLDN18, LY6G6D, GPR56, CD276, TPBG, MST1R, CDH3, EpCAM, TNFRSF10B, PD-L1, TROP2, EphA2 and CD71.

[0082] The present disclosure also provides a method for degrading cMET protein on target cells. The method utilizes a binding agent that specifically binds to: (1) an extracellular epitope on the cMET protein; and (2) an extracellular epitope on a membrane-associated degradation protein on the target cell. The binding agent comprises a first binding domain that selectively binds to the cMET protein and a second binding domain that selectively binds to the membrane-associated degradation protein. Simultaneous binding of the binding agent to the cMET protein and the membrane-associated degradation protein results in degradation of both the cMET protein and the membrane-associated degradation protein.

[0083] In some embodiments, the membrane-associated degradation protein is a cell surface protein that is degraded after a binding agent (e.g., an antibody) binds to the protein. In some embodiments, the membrane-associated degradation protein is RNF43, ZHFR3, RNF167, RNF128, and RNF130.

[0084] In one aspect, the present disclosure provides a method for degrading cMET protein on target cells, the method comprising:

[0085] contacting the cMET protein and the membrane-associated internalization protein on the target cell with a binding agent, wherein contacting the cMET protein and the membrane-associated internalization protein with the binding agent results in internalization and degradation of the cMET protein; and

[0086] wherein the binding agent comprises: (a) a first binding domain that specifically binds to an extracellular epitope on a membrane-associated internalization protein; and (b) a second binding domain that specifically binds to an extracellular epitope on a cMET protein;

[0087] Among them, membrane-associated internalization proteins are selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, AD AM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, CD276, TPBG, MST1R, CDH3, EpCAM, TNFRSF10B, PD-L1, TROP2, EphA2 and CD71. In some embodiments, the binding agent comprises an antibody. In some embodiments, the binding agent comprises a multispecific antibody. In some embodiments, the binding agent comprises a bispecific antibody. In some embodiments, the binding agent comprises an IgG antibody. In some embodiments, the binding agent comprises an IgG antibody. In some embodiments, the binding agent comprises a knob-and-hole bispecific IgG. In some embodiments, the binding agent is not an antibody drug conjugate ("ADC"). In some embodiments, the binding agent comprises a bispecific binding agent. In some embodiments, the binding agent comprises a bispecific antibody. In some embodiments, the binding agent comprises a bispecific diabody. In some embodiments, the binding agent comprises a bispecific Fab2. In some embodiments, the binding agent comprises a bispecific camelid antibody. In some embodiments, the binding agent comprises a bispecific peptide antibody scFv-Fc. In some embodiments, the binding agent comprises an Fc-Fab. In some embodiments, the binding agent comprises a knob-in-hole bispecific Fc-Fab.

[0088] binder

[0089] The binding agents of the present disclosure contain two binding domains: one is specific for a degradation protein and the other is specific for a cMET protein. The binding agents of the present disclosure include, but are not limited to, agents in which the degradation protein binding domain and the cMET binding domain are each independently selected from an antibody (or half of an antibody), a nanobody or a minibody, a Fab fragment, a single-chain variable fragment (scFv), and a single-domain antibody (sdAb) or a functional fragment thereof. The two binding domains can be molecules of the same type or different types. For example, the binding agents of the present disclosure include, but are not limited to, binding agents having an IgG that binds to a degradation protein and a scFv domain that binds to cMET. The two binding domains of the binding agent can be connected by a covalent bond, a non-covalent interaction, or a combination thereof.

[0090] Binders can generally be in the form of proteins, glycoproteins, lipoproteins, phosphoproteins, and the like. Some binding agents disclosed herein are in the form of antibodies or antibody derivatives. In some embodiments, the target protein binding domain is selected from a half antibody, a nanobody or a mini antibody, a F(ab')2 fragment, a Fab fragment, a single-chain variable fragment (scFv), and a single domain antibody (sdAb) or a functional fragment thereof. The two binding domains can be in the form of bispecific antibodies, bispecific diabodies, bispecific camel antibodies, or bispecific peptide antibodies. Antibody derivatives do not have to be derived from a specific wild-type antibody. For example, known techniques (such as phage display, etc.) can be used to produce and select small proteins having binding domains similar to antibody complementary determining regions (CDRs). In some embodiments, the antibody binding portion includes scFv. The binding domain can also be derived from a natural or synthetic ligand or receptor that specifically binds to the cMET protein, whether soluble or membrane-bound.

[0091] Bispecific antibodies can be prepared by known methods. Embodiments of the present disclosure include "knob-into-hole" bispecific antibodies, in which the originally symmetrical dimerization region of the bispecific binding agent is altered to make it asymmetric. For example, a knob-into-hole bispecific IgG with specificity for antigens A and B can be altered so that the Fc portion of the A binding chain has one or more protrusions ("knob"), and the Fc portion of the B binding chain has one or more depressions ("hole"), wherein the knob and hole are arranged to interact. This reduces homodimerization (AA and BB antibodies) and promotes the heterodimerization required for the bispecific binding agent. See, for example, Y.Xu et al., mAbs (2015) 7(1): 231-42. In some embodiments, the bispecific binding agent has a knob-into-hole design. In some embodiments, the "knob" comprises a T336W change in the CH3 domain, i.e., the threonine at position 336 is replaced with a tryptophan. In some embodiments, the "hole" comprises one or a combination of T366S, L368A, and Y407V. In some embodiments, the "hole" comprises T366S, L368A, and Y407V.

[0092] In some embodiments, the binding agent comprises an FcRn receptor recognition domain to facilitate return of the binding agent to the extracellular space if the binding agent is internalized.

[0093] In another aspect, the present disclosure provides a binding agent comprising an antibody or antibody derivative, the binding agent comprising:

[0094] a) a first binding domain that specifically binds to an extracellular epitope of a cMET protein of a target cell; and

[0095] b) a second binding domain that specifically binds to an extracellular epitope of the membrane-associated internalization protein on the target cell;

[0096] The membrane-associated internalization protein is selected from CD205, CD166, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56 and CD71.

[0097] Degraded proteins

[0098] The methods and binding agents disclosed herein can utilize membrane-associated degradation proteins to cause degradation of cMET proteins. The disclosure can utilize membrane-associated degradation proteins to cause ubiquitination after binding of the binding agent to the membrane-associated degradation protein. By also binding to cMET at the first binding domain and using the second binding domain to bind to the membrane-associated degradation protein, the multifunctional binding agent can cause degradation of cMET protein along with the membrane-associated degradation protein.

[0099] The membrane-associated degradation proteins used in the methods and bifunctional binding agents of the present disclosure may include cell surface proteins that are degraded after the binding agent (e.g., antibody) binds to and / or internalizes the protein. Such membrane-associated degradation proteins may include cell surface proteins targeted by antibody drug conjugates, which may rely on the degradation of the antibody-protein complex to ensure the release of the conjugated drug. Examples of such membrane-associated degradation proteins used in the methods of the present disclosure may include, for example, TROP2. In some embodiments, the membrane-associated degradation protein is an E3 ligase. In some embodiments, the membrane-associated degradation protein is RNF43 (i.e., RING finger protein 43).

[0100] Degradation proteins for the methods and bifunctional binding agents of the present disclosure may include cell surface proteins that are internalized after the binding agent (e.g., antibody) binds to the protein. Such membrane-associated internalization proteins include cell surface proteins currently targeted by antibody drug conjugates, which typically rely on the internalization of the antibody protein complex to ensure the release of the conjugated drug. Examples of such membrane-associated internalization proteins for the methods of the present disclosure include, for example, CEACAM5 (i.e., CEA cell adhesion molecule 5), CEACAM6 (i.e., CEA cell adhesion molecule 6), HER3 (i.e., receptor tyrosine protein kinase erbB-3), MUC1 (i.e., mucin 1), CD205 (i.e., lymphocyte antigen 75), CD166 (i.e., activated leukocyte cell adhesion molecule, also known as ALCAM), PRLR (i.e., prolactin receptor), SLC34A2 (i.e., solute carrier family 34 member 2), ITGB6 (i.e., integrin subunit β6), LRRC15 (i.e., containing rich Leucine repeat protein 15), MUC16 (i.e., mucin 16), SLC39A6 (i.e., solute carrier family 39 member 6), AXL (i.e., AXL receptor tyrosine kinase), MMP14 (i.e., matrix metalloproteinase 14), CD40 (i.e., cluster of differentiation 40), CD228A (i.e., melanin transferrin), CD70 (i.e., cluster of differentiation 70), MUC5A (i.e., mucin 5A), CD44 (i.e., homing cell adhesion molecule), ITGB1 (i.e., integrin beta-1), STn (e.g., carbohydrate antigen STn), KAAG1 (i.e., kidney-associated antigen 1), DL K1 (i.e., delta-like non-canonical Notch ligand 1), 5T4 (i.e., carcinoembryonic antigen 5T4), SEZ6 (i.e., epilepsy-associated 6 homolog), CD123 (i.e., interleukin 3 receptor), ADAM9 (i.e., a disintegrin and metalloproteinase 9), I-Ag7 (i.e., MHC class II molecule Ag7), ENPP3 (i.e., ectonucleotide pyrophosphatase / phosphodiesterase 3), CD37 (i.e., tetraspanin CD37), CD46 (i.e., CD46 complement regulatory protein), CD56 (i.e., neural cell adhesion molecule), CD74 (i.e., invariant chain of MHC II), IGF1R (i.e., insulin-like growth factor receptor 1), 1 receptor), ROR1 (i.e., receptor tyrosine kinase-like orphan receptor 1), CDH6 (i.e., cadherin 6), ROR2 (i.e., receptor tyrosine kinase-like orphan receptor 2), GPR20 (i.e., G protein-coupled receptor 20), TM4SF1 (i.e., transmembrane 4L size family member 1), B7-H4 (i.e., V-Set domain-containing inhibitor of T cell activation 1), ALPP (i.e., placental alkaline phosphatase), LY6E (i.e., lymphocyte antigen 6 family member E), CLDN18 (i.e., claudin 18), LY6G6D (i.e., lymphocyte antigen 6 family member G6D), GPR56 (i.e.,Adhesion G protein-coupled receptor G1), CDH3 (chromodomain helicase DNA binding protein 3), CD276 (cluster of differentiation 276), TROP2 (trophoblast cell surface antigen 2), TNFRSF10B (TNF receptor superfamily member 10), PD-L1 (programmed death ligand 1), EpCAM (epithelial cell adhesion molecule), TPBG (trophoblast glycoprotein), EGFR (epithelial growth factor receptor), MST1R (macrophage stimulating 1 receptor), EphA2 (ephrin receptor A2), and CD71 (transferrin receptor-1). It has been shown that these proteins are internalized into cells after binding of binding agents (e.g., antibodies) to the extracellular epitopes of the proteins.

[0101] In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123 , ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, L Y6E, CLDN18, LY6G6D, GPR56, CD276, TPBG, MST1R, CDH3, EpCAM, TNFRSF10B, PD-L1, TROP2, EphA2 and CD71. In some embodiments, the membrane-associated internalization protein is selected from CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.

[0102] In some embodiments, the membrane-associated internalization protein is selected from the group consisting of CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.

[0103] In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, CD40, CD228A, CD70, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, CD123, ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, and B7-H4.

[0104] In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CD205, CD166, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.

[0105] In some embodiments, the membrane-associated internalization protein is selected from SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CD205, CD166, LRRC15, MUC16, SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.

[0106] In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from the group consisting of CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.

[0107] In some embodiments, the membrane-associated internalization protein is selected from CD205, CD166, CD40, CD70, CD44, CD123, CD37, CD228, CD46, CD56, CD74, CDH6, and CD71. In some embodiments, the membrane-associated internalization protein is selected from SLC34A2, ITGB6, LRRC15, MUC16, AXL / UFO, MMP14, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, IGF1R, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from CEACAM5, CEACAM6, HER3, MUC1, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MMP14, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, IGF1R, ROR1, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56 and CD71.

[0108] In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL, CD40, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71.

[0109] In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL / UFO, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, LY6E, CLDN18, LY6G6D, GPR56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from the group consisting of SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, and ALPP.

[0110] In some embodiments, the membrane-associated internalization protein is selected from CD205, CD166, CD40, CD228, CD46, CD56, and CD71. In some embodiments, the membrane-associated internalization protein is selected from SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, and GPR56.

[0111] In some embodiments, the membrane-associated internalization protein is CDH3. In some embodiments, the membrane-associated internalization protein is MUC1. In some embodiments, the membrane-associated internalization protein is CD276. In some embodiments, the membrane-associated internalization protein is TROP2. In some embodiments, the membrane-associated internalization protein is CD71. In some embodiments, the membrane-associated internalization protein is HER3. In some embodiments, the membrane-associated internalization protein is TNFRSF10B. In some embodiments, the membrane-associated internalization protein is ITGB6. In some embodiments, the membrane-associated internalization protein is PD-L1. In some embodiments, the membrane-associated internalization protein is EpCAM. In some embodiments, the membrane-associated internalization protein is TPBG. In some embodiments, the membrane-associated internalization protein is EGFR. In some embodiments, the membrane-associated internalization protein is MST1R. In some embodiments, the membrane-associated internalization protein is EphA2. In some embodiments, the membrane-associated internalization protein is ADAM9. In some embodiments, the membrane-associated internalization protein is IGF1R.

[0112] Degradation proteins for use in the methods and bifunctional binding agents of the present disclosure may include cell surface proteins that are degraded after the binding agent (e.g., antibody) binds to the protein. Such membrane-associated degradation proteins include cell surface proteins currently targeted by antibody drug conjugates, which typically rely on the degradation of antibody protein complexes to ensure the release of conjugated drugs. Examples of such membrane-associated degradation proteins for use in the methods of the present disclosure include, for example, RNF43 (i.e., RING finger protein 43), RNF128 (i.e., RING finger protein 128), RNF130 (i.e., RING finger protein 130), and ZNRF3 (i.e., zinc finger and RING finger 3).

[0113] In some embodiments, the membrane-associated degradation protein is RNF43. In some embodiments, the membrane-associated degradation protein is RNF128. In some embodiments, the membrane-associated degradation protein is RNF130. In some embodiments, the membrane-associated degradation protein is ZNRF3.

[0114] First binding region

[0115] In some embodiments, the first binding domain is derived from an antibody against a membrane-associated internalization protein. Such antibodies are known to those skilled in the art and can be incorporated into the methods and binding agents of the present disclosure. For example, in some embodiments, the complementary determining regions ("CDRs") of known antibodies against a membrane-associated internalization protein of interest can be incorporated into the binding agents and methods of the present disclosure using known techniques. Exemplary antibodies suitable for incorporation into the methods and binding agents of the present disclosure include those described below.

[0116] For example, antibodies targeting CEACAM5 are known in the art, including, for example, the CC4 antibody, which is disclosed in, for example, Zheng, Chaogu et al., "A novel anti-CEACAM5 monoclonal antibody, CC4, suppresses colorectal tumor growth and enhances NK cells-mediated tumor immunity." PloS one 6.6 (2011): e21146. Additional antibodies targeting CEACAM5 suitable for use in the present disclosure include, for example, the anti-CEACAM5 antibodies MN-14, MN-15, and MN-3, which are described in, for example, Blumenthal, Rosalyn D. Hans J. Hansen, and David M. Goldenberg, "Inhibition of adhesion, invasion, and metastasis by antibodies targeting CEACAM6 (NCA-90) and CEACAM5 (Carcinoembryonic Antigen)." Cancer research 65.19 (2005): 8809-8817.

[0117] Antibodies targeting CEACAM6 are known in the art, including, for example, anti-CEACAM6 antibodies sdAb, 2Ab, and 4Ab, which are described in, for example, the following literature: Wu, Shang-Jung et al., "Migration and invasion of NSCLC suppressed by the downregulation of Src / focal adhesion kinase using single, double and tetra domain anti-CEACAM6 antibodies." Translational oncology 14.7(2021):101057. Additional antibodies targeting CEACAM6 suitable for use in the present disclosure include, for example, the anti-CEACAM6 antibodies MN-3 and MN-15, which are described, for example, in Blumenthal, Rosalyn D. Hans J. Hansen, and David M. Goldenberg, "Inhibition of adhesion, invasion, and metastasis by antibodies targeting CEACAM6 (NCA-90) and CEACAM5 (Carcinoembryonic Antigen)." Cancer research 65. 19 (2005): 8809-8817.

[0118] Antibodies targeting HER3 (also known as ErbB-3) are known in the art, including, for example, the anti-HER3 antibody GSK2849330, which is described, for example, in the following literature: Gan, Hui K. et al., "Aphase I, first-in-human study of GSK2849330, an anti-HER3 monoclonal antibody, in HER3-expressing solid tumors." The oncologist 26.10(2021):e1844-e1853. Further anti-HER3 antibodies include, for example, Patritumab (U3-1287), which is described in, for example, Hashimoto, Yuuri et al., "A Novel HER3-Targeting Antibody–Drug Conjugate, U3-1402, Exhibits Potent Therapeutic Efficacy through the Delivery of Cytotoxic Payload by Efficient Internalization Preclinical Evaluation of U3-1402, a HER3-Targeting ADC." Clinical Cancer Research 25.23(2019):7151-7161.

[0119] Antibodies targeting MUC1 are known in the art, including, for example, anti-MUC1 antibodies MY.1E12, KL6, 5E5, and TAB004, which are described in the following literature: Bose, Mukulika and Pinku Mukherjee, "Potential of anti-MUC1 antibodies as a targeted therapy for gastrointestinal cancers." Vaccines 8.4 (2020): 659.

[0120] Antibodies targeting CD205 are known in the art, including, for example, the anti-CD205 antibody MEN1309 / OBT076, which is described, for example, in Rieke, Damian T. and Ulrich Keller, "ACD205-directed antibody drug conjugate–lymphoma precision oncology or sophisticated chemotherapy?" Haematologica 105.11(2020):2504.

[0121] Antibodies targeting CD166 are known in the art, for example, the anti-CD166 antibody CX-2009, which is described in, for example, Boni, Valentina et al., "Praluzatamab ravtansine, a CD166-targeting antibody-drug conjugate, in patients with advanced solid tumors: an open-label phase 1 / 2 trial of Praluzatamab ravtansine in patients with advanced tumors." Clinical Cancer Research (2022).

[0122] Antibodies targeting PRLR are known in the art, for example, anti-PRLR antibody ABBV-176, which is described in, for example, the following literature: Anderson, Mark G. et al., "ABBV-176, a PRLR antibody drug conjugate with apotent DNA-damaging PBD cytotoxin and enhanced activity with PARPinhibition." BMC cancer 21.1(2021): 1-11.]). Additional antibodies targeting CEACAM6 suitable for use in the present disclosure include, for example, anti-CEACAM6 antibody LFA102, which is described in, for example, Damiano, Jason S. et al., "Neutralization of Prolactin Receptor Function by Monoclonal Antibody LFA102, a Novel Potential Therapeutic for the Treatment of Breast Cancer Preclinical Development of Anti-PRLR Antibody LFA102." Molecular cancer therapeutics 12.3(2013): 295-305.

[0123] Antibodies targeting SCL34A2 are known in the art, for example, anti-NaPi2b antibodies, which are described in the following literature: Lin, Kedan et al., "Preclinical Development of an Anti-NaPi2b (SLC34A2) Antibody–Drug Conjugate as a Therapeutic for Non–Small Cell Lung and Ovarian Cancers Preclinical Development of NaPi2b Antibody–Drug Conjugate." Clinical Cancer Research 21.22 (2015): 5139-5150. Additional antibodies suitable for incorporation into the binding agents of the present disclosure include the anti-SCL34A2 antibody MX35, which is described in the following literature: Yin, Beatrice WT et al., "Monoclonal antibody MX35 detects the membrane transporter NaPi2b (SLC34A2) in human carcinomas." Cancer immunity 8.1 (2008).

[0124] Antibodies targeting ITGB6 are known in the art, including, for example, the antibody SGN-B6A, which is described in, for example, Patnaik, Amita et al., "A phase 1 study of SGN-B6A, an antibody-drug conjugate targeting integrin beta-6, in patients with advanced solid tumors (SGN-B6A-001, Trial in Progress)." (2021). Another antibody suitable for incorporation into the present disclosure includes the anti-ITGB6 antibody TPS3144-TPS3144, which is described in, for example, Zheng, Xiaoxia et al., "Silencing of ITGB6 inhibits the progression of cervical carcinoma via regulating JAK / STAT3 signaling pathway." Annals of Translational Medicine 9.9 (2021).

[0125] Antibodies targeting LRRC15 are known in the art, including, for example, the anti-LRCC15 antibody ABBV-085, which is described in, for example, the following literature: Demetri, George D. et al., "First-in-Human Phase I Study of ABBV-085, an Antibody–Drug Conjugate Targeting LRRC15, in Sarcomas and Other Advanced Solid Tumors Phase I Study of ABBV-085, an LRRC15-Targeting ADC." Clinical Cancer Research 27.13(2021):3556-3566; and Slemmons, Katherine K. et al., "LRRC15 antibody-drug conjugates show promise as osteosarcoma therapeutics in preclinical studies." Pediatric blood & cancer 68.2(2021):e28771]).

[0126] Antibodies targeting MUC16 are known in the art and include, for example, the anti-MUC16 antibody OC125, which is described, for example, in Rao, Thapi Dharma et al., "Novel monoclonal antibodies against the proximal (carboxy-terminal) portions of MUC16." Applied immunohistochemistry & molecular morphology: AIMM / official publication of the Society for Applied Immunohistochemistry 18.5 (2010): 462. Additional anti-MUC16 antibodies include, for example, those described in Aithal, Abhijit et al., "MUC16 as a novel target for cancer therapy." Expertopinion on therapeutic targets 22.8 (2018): 675-686; and Rao, Thapi Dharma et al., "Antibodies against specific MUC16 glycosylation sites inhibit ovarian cancer growth." ACS chemical biology 12.8 (2017): 2085-2096].

[0127] Antibodies targeting SLC39A6 are known in the art, including, for example, anti-SLC39A6 antibodies described in Cui, Shen et al., "SLC39A6: a potential target for diagnosis and therapy of esophageal carcinoma." Journal of Translational Medicine 13(2015): 321. Additional anti-SLC29A6 antibodies include, for example, those described in Sussman, Smith et al., "SGN-LIV1A: A novel antibody-drug conjugate targeting LIV-1 for the treatment of metastatic breast cancer." Mol Chancer Ther (2014) 13(12): 2991-3000; and Wan and Wang, "Role of SLC39A in the development and progression of liver cancer." Oncology Letters 23.3.(2022): 77.

[0128] Antibodies targeting AXL are known in the art, including, for example, AXL-specific antibodies described in the following literature: Vajkoczy, Knyazev et al., "Dominant-negative inhibition of the Axl receptor tyrosine kinase suppresses brain tumor cell growth and invasion and prolongs survival." Proceedings of the National Academy of Sciences 103.15(2006):5799-5804. Additional anti-AXL antibodies include, for example, the anti-AXL antibody 20G7-D9, described in the following literature: Leconet, Chentouf et al., "Therapeutic activity of anti-AXL antibody against triple-negative breast caser patient-derived xenografts and metastasis." Clin Cancer Research 23.11(2017):2806-2816.

[0129] Antibodies targeting CD40 are known in the art, including, for example, anti-CD40 antibodies, which are described in the following literature: Xu, Gao et al., "Repulsive guidance molecule a blockade exerts the immunoregulatory function in DCs stimulated with ABP and LPS." Human vaccines & immunotherapeutics 12.8 (2016): 2169-2180. Additional anti-CD40 antibodies include, for example, those described in: Silvin, Chapuis et al., "Elevated calprotectin and abnormal myeloid cell subsets discriminate severe from mild COVID-19." Cell 182.6(2020):1401-1418; and Ceglia, Zurawski et al., "Anti-CD40 Antibody Fused to CD40Ligand Is a Superagonist Platform for Adjuvant Intrinsic DC-Targeting Vaccines." Frontiers in immunology 12:786144(2021).

[0130] Antibodies targeting CD228 are known in the art, including, for example, anti-MELTF antibodies, which are described in the following literature: Sawaki, Kanda et al., "Level of melanotransferrin in tissue and sera serves as aprognostic marker of gastric cancer." Anticancer Research 39.11(2019):6125-6133. Additional anti-CD228 antibodies include, for example, those described in the following literature: Singh, Eyford et al., "Discovery of a Highly Conserved Peptide in the Iron Transporter Melanotransferrin that Traverses an Intact Blood Brain Barrier and Localizesin Neural Cells." Frontiers in neuroscience 15:596976.(2021):473.

[0131] Antibodies targeting MUC5A are known in the art and include, for example, the anti-MUC5A antibody MUC5:TR-3A, which is described in: Zuhdi Alimam, Piazza et al., "Muc-5 / 5ac mucin messenger RNA and protein expression is a marker of goblet cell metaplasia in murine airways." American journal of respiratory cell and molecular biology 22.3 (2000): 253-260. Additional anti-MUC5 antibodies include, for example, those described in Wang, Jin et al., "Expression of survivin, MUC2 and MUC5 in colorectal cancer and their association with clinical pathological characteristics." Oncology Letters 14.1 (2017): 1011-1016; and Reis, David et al., "Immunohistochemical study of MUC5AC expression in human gastric carcinomas using a novel monoclonal antibody." International journal of cancer 74.1 (1997): 112-121.

[0132] Antibodies targeting ITGB1 are known in the art, including, for example, anti-ITGB1 antibodies, which are described in the following literature: Du, Yang et al., "The circular RNAcircSKA3 binds integrinβ1to induce invadopodiumformation enhancing breast cancer invasion." Molecular Therapy 28.5(2020):1287-1298. Additional anti-ITGB1 antibodies include, for example, those described in the following literature: Kawahara, Niwa et al., "Integrinβ1is an essential factor in vasculogenic mimicry of human cancer cells." Cancer science 109.8(2018):2490-2496; and Wang and Li, "Ropivacaineinhibits the proliferation and migration of colorectal cancer cells throughITGB1." Bioengineered 12.1(2021):44-53.

[0133] Antibodies targeting STn are known in the art, including, for example, anti-STn antibodies described in Prendergast, da Silva et al., "Novel anti-Sialyl-Tn monoclonal antibodies and antibody-drug conjugates demonstrate tumor specificity and anti-tumor activity." mAbs 9, 4 (2017): 615-627. Additional anti-STn antibodies include, for example, those described in Eavarone, David A et al., "Humanized anti-Sialyl-Tn antibodies for the treatment of ovarian carcinoma." PloS one 13, 7 (2018) e0201314.27.

[0134] Antibodies targeting KAAG1 are known in the art and include, for example, the anti-KAAG1 antibody anti-KAAG1 AB-3A, which is described in US Pat. No. 9,393,302 B2.

[0135] Antibodies targeting DLK1 are known in the art, including, for example, the anti-DLK1 antibody anti-DLK1SIP (EB3), which is described in Bujak, Ritz et al., "A monooclonal antibody to human Dlk1 reveals differential expression in cancer and absence in healthy tissues." Antibodies 4.2 (2015): 71-87. Additional anti-DLKL antibodies include, for example, those described in Takagi, Zhao et al., "Delta-like 1 homolog (DLK1) as a possible therapeutic target and its application to radioimmunotherapy using 125I-labelled anti-DLK1 antibody in lung cancer models (HOT1801 and FIGHT004). " Lung Cancer 153 (2021): 134-142; and Huang, Zhang et al., "Up-regulation of DLK1 as an imprinted gene could contribute to human hepatocellular carcinoma." Carcinogenesis 28.5 (2007): 1094-1103.

[0136] Antibodies targeting 5T4 are known in the art, including, for example, the anti-5T4 antibody anti-5T4 IgG1, which is described in Shapiro, Vaishampayan et al., "First-in-human trial of an anti-5T4 antibody-monomethylauristatin conjugate, PF-06263507, in patients with advanced solid tumors." Investigational New Drugs 35.3(2017):315-323. Additional anti-5T4 antibodies include, for example, those described in Owens, Sheard et al., "Preclinical assessment of CART-cell therapy targeting the tumor antigen 5T4 in ovarian cancer." Journal of Immunotherapy 41.3(2018):130-140.

[0137] Antibodies targeting SEZ6 are known in the art, including, for example, anti-SEZ6 antibodies described in the following literature: Jiang, Chen et al., "Correlation between human seizure-related gene 6 variants and idiopathic generalized epilepsy in a Southern Chinese Han population." Neural Regeneration Research 7.2 (2012): 96-100. Additional anti-SEZ6 antibodies include, for example, those described in the following literature: Kuhn, Koroniak et al., "Secretome protein enrichment identifies physiological BACE1 protease substrates in neurons." The EMBO journal 31.14 (2012): 3157-3168.

[0138] Antibodies targeting ADAM9 are known in the art, including, for example, anti-ADAM9 antibodies, which are described in: Mazzocca, Coppari et al., "Asecreted form of ADAM9 promotes carcinoma invasionthrough tumor-stromal interactions." Cancer research 65.11(2005):4728-4738. Additional anti-ADAM9 antibodies include, for example, those described in: Zigrino, Mauch et al., "Adam-9expression and regulation in human skin melanoma and melanoma cell lines." International journal of cancer 116.6(2005):853-859; and Kim, Jeung et al., "The Effect of Disintegrin–Metalloproteinase ADAM9 in Gastric Cancer Progression." Molecular cancer therapeutics 13.12(2014):3074-3085.

[0139] Antibodies targeting I-Ag7 are known in the art and include, for example, anti-I-Ag7 antibodies described in Zhang, Crawford et al., "Monoclonal antibody blocking the recognition of aninsulin peptide-MHC complex modulates type 1 diabetes." Proceedings of the National Academy of Sciences 111.7 (2014): 2656-2661. Additional antibodies targeting I-Ag7 include, for example, those described in Noorchashm, Hooman et al., "I-Ag7-mediated antigen presentation by B lymphocytes is critical in overcoming a checkpoint in T cell tolerance to islet β cells of nonobese diabetic mice." The Journal of Immunology 163.2 (1999): 743-750; and Gardiner, Richards et al., "Conformation of MHC class II I-Ag7 is sensitive to the P9 anchor amino acid in bound peptide." International immunology 19.9 (2007): 1103-1113.

[0140] Antibodies targeting ENPP3 are known in the art, including, for example, anti-ENPP3 antibodies described in the following literature: Boggavarapu, Lalitkumar et al., "Compartmentalized gene expression profiling ofreceptive endometrium reveals progesterone regulated ENPP3 is differentially expressed and secreted in glycosylated form." Scientific reports 6.1(2016): 1-13. Additional anti-ENPP3 antibodies include, for example, those described in the following literature: Schiechl, Hermann et al., "Basophils trigger fibroblast activation in cardiac allograft fibrosis development." American Journal of Transplantation 16.9(2016): 2574-2588.

[0141] Antibodies targeting CD46 are known in the art and include, for example, the anti-CD46 antibody anti-CD46 antibody YS5, which is described in: Su, Liu et al., "Targeting CD46 for both adenocarcinoma and neuroendocrine prostate cancer." JCI insight 3.17 (2018) e121497. Additional anti-CD46 antibodies include, for example, those described in: Carver-Ward, Hollanders et al., "Progesterone does not potentiate the acrosome reaction in human spermatozoa: flowcytometric analysis using CD46 antibody." Human reproduction 11.1 (1996): 121-126; and Krey, Himmelreich et al., "Function of bovine CD46 as a cellular receptor for bovine viral diarrhea virus is determined by complement control protein 1." Journal of virology 80.8 (2006): 3912-3922.

[0142] Antibodies targeting CD56 are known in the art, including, for example, anti-CD56 antibodies, which are described in the following literature: Silvin, Chapuis et al., "Elevated calprotectin and abnormal myeloid cell subsets discriminate severe from mild COVID-19." Cell 182.6(2020):1401-1418. Additional anti-CD46 antibodies include, for example, those described in the following literature: Zhan, Guo et al., "Glioma stem-like cells evade interferon suppression through MBD3 / NuRD complex-mediated STAT1 downregulation." The Journal of experimental medicine 217, 5(2020):e20191340; and Feng, Wang et al., "Differential killing of CD56-expressing cells by drug-conjugated human antibodies targeting membrane-distal and membrane-proximal non-overlapping epitopes." mAbs 8.4(2016):799-810.

[0143] Antibodies targeting ROR1 are known in the art, including, for example, the anti-ROR1 antibody anti-ROR14A5, which is described in Balakrishnan, Goodpaster et al., "Analysis of ROR1 Protein Expression in Human Cancer and Normal Tissues." Clinical Cancer Research 23.12 (2017): 3061-3071. Additional anti-ROR1 antibodies include, for example, those described in Baskar, Wiestner et al., "Targeting malignant B cells with an immunotoxin against ROR1." mAbs.4.3 (2012) 349-361; and Zhang, Chen et al., "ROR1 is expressed in human breast cancer and associated with enhanced tumor-cell growth." PloS one 7, 3 (2012): e31127.

[0144] Antibodies targeting GPR20 are known in the art, including, for example, anti-GPR20 antibodies described in Wheway, Schmidts et al., "An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes." Nature cell biology 17, 8 (2015): 1074-1087. Additional anti-GPR20 antibodies include, for example, those described in Iida, Ahmed et al., "Identification and Therapeutic Targeting of GPR20, Selectively Expressed in Gastrointestinal Stromal Tumors, with DS-6157a, a First-in-Class Antibody–Drug Conjugate." Cancer Discovery 11.6 (2021): 1508-1523.

[0145] Antibodies targeting TM4SF1 are known in the art, including, for example, anti-TM4SF1 antibodies described in Zacharias, Frank et al., “Regeneration of the lung alveolus by anevolutionarily conserved epithelial progenitor.” Nature 555, 7695 (2018): 251-255. Additional antibodies targeting TM4SF1 include, for example, the anti-TM4SF1 antibody 8G4, which is described in the following literature: Lin, Merley et al., “TM4SF1: a new vascular t9herapeutic target in cancer.” Angiogenesis 17, 4 (2014): 897-907; and anti-TM4SF1 antibodies, which are described in the following literature: Wang, Sun et al., “B7-H3 suppresses doxorubicin-induced senescence-like growth arrest in colorectal cancer through the AKT / TM4SF1 / SIRT1 pathway” Cell death & disease 12, 5 (2021): 453.

[0146] Antibodies targeting B7-H4 are known in the art, including, for example, anti-B7-H4 antibodies described in the following literature: Podojil, Glaser et al., "Antibody targeting of B7-H4 enhances the immune response inurothelial carcinoma." Oncoimmunology 9, 1(2020): 1744897. Additional antibodies targeting B7-H4 include, for example, those described in Miao and Sun, "Development of a novel anti-B7-H4 antibody enhances anti-tumor immune response of human T cells." Biomedicine & pharmacology 141 (2021): 111913; and Dangaj, Lanitis et al., "Novel Recombinant Human B7-H4 Antibodies Overcome Tumoral Immune Escape to Potentiate T-Cell Antitumor Responses Overcoming B7-H4–Mediated T-Cell Inhibition." Cancer research 73.15 (2013): 4820-4829.

[0147] Antibodies targeting ALPP are known in the art, including, for example, the anti-ALPP antibody anti-ALPP SP15, which is described in the following literature: Zwolanek, Satue et al., "Tracking mesenchymal stem cell contributions to regeneration in an immunocompetent cartilage regeneration model." JCI insight 2.20 (2017) e87322. Additional antibodies targeting ALPP include, for example, those described in the following literature: Chen, Chen et al., "Placental alkaline phosphatase promotes Zika virus replication bystabilizing viral proteins through BIP." MBio 11.5 (2020): e01716-20.; and Egerbacher et al., "Hematopoietic bone marrow cells participate inendothelial, but not epithelial or mesenchymal cell renewal in adult rats." Journal of cellular and molecular medicine 15.10(2011):2232-2244.

[0148] Antibodies targeting LY6E are known in the art, including, for example, anti-LY6E antibodies, which are described in the following literature: Mar, Rinkenberger et al., "LY6E mediates an evolutionarily conserved enhancement of virus infection by targeting a late entry step." Nature communications 9.1(2018):1-14. Additional antibodies targeting LY6E include, for example, the anti-LY6E antibody anti-LY6E MTS35, which is described in the following literature: Langford, Outhwaite et al., "Deletion of the Syncytin Areceptor Ly6e impairssyncytiotrophoblast fusion and placental morphogenesis causing embryoniclethality in mice." Scientific reports 8, 1 (2018): 3961.; and the anti-LY6E antibody anti-LY6E9B12, which is described in the following literature: Dela Cruz Chuh, Josefa et al., "Preclinical optimization of Ly6E-targeted ADCs for increased durability and efficacy of anti-tumor response." MAbs 13.1 (2021).

[0149] Antibodies targeting CLDN18 are known in the art, including, for example, anti-CLDN18 antibodies described in the following literature: Türeci, Mitnacht-Kraus et al., "Characterization of zolbetuximab in pancreatic cancer models." Oncoimmunology 8.1 (2019): e1523096. Additional anti-CLDN18 antibodies include, for example, those described in the following literature: Matsusaka, Ushiku et al., "Coupling CDH17 and CLDN18 markers for comprehensive membrane-targeted detection of human gastric cancer." Oncotarget 7, 39 (2016): 64168-64181.

[0150] Antibodies targeting LY6G6D are known in the art, including, for example, anti-LY6G6D antibodies described in Sewda, Coppola et al., “Cell-surface markers for colon adenoma and adenocarcinoma.” Oncotarget 7, 14 (2016): 17773-89. Additional anti-LY6G6D antibodies include, for example, the anti-LY6G6D antibody anti-LY6G6D clone 10C1, which is described in: Corrales, Hipp et al., "LY6G6D is a selectively expressed colorectal cancer antigen that can be used for targeting a therapeutic T-cell response by a T-cell engager." Frontiers inimmunology 13(2022):1008764.; and anti-LY6G6D antibodies described in Wang, Sun et al., "Novel Anti-LY6G6D / CD3 T Cell-Dependent Bispecific Antibody for the Treatment of Colorectal Cancer." Molecular Cancer Therapeutics 21:6(2022):974-985.

[0151] Antibodies targeting GPR56 are known in the art, including, for example, the anti-GPR56 antibody anti-GPR56 10C7, which is described in the following document: Chatterjee, Zhang et al., "Anti-GPR56 monoclonal antibody potentiates GPR56-mediated Src-Fak signaling to modulate cell adhesion." Journal of Biological Chemistry 296(2021)100261. Additional anti-GPR56 antibodies include, for example, those described in Iguchi, Sakata et al., "Orphan G protein-coupled receptor GPR56 regulates neural progenitor cell migration via a Gα12 / 13and Rho pathway." Journal of Biological Chemistry 283.21 (2008): 14469-14478.; and Chen, Yang et al., "GPR56 is essential for testis development and male fertility in mice." Developmental Dynamics 239.12 (2010): 3358-3367.

[0152] Antibodies targeting MMP14 are known in the art, including, for example, anti-MMP14 antibodies described in the following literature: Zhang, Zhang et al., "MMP-14 aggravates onset of severe preeclampsia by mediating soluble endoglin release." European review for medical and pharmacological sciences 22, 5 (2018): 1209-1215. Additional anti-MMP14 antibodies include, for example, those described in the following literature: Fischer and Riedl, "Inhibitory antibodies designed for matrixmetalloproteinase modulation." Molecules 24.12 (2019): 2265.

[0153] Antibodies targeting cMET are known in the art, including, for example, anti-cMET antibodies, which are described in, for example, Lee, D. et al., "Development of antibody-based c-Met inhibitors for targeted cancer therapy." ImmunoTargets and therapy 4 (2015): 35-44. Additional anti-cMET antibodies include those described in, for example, Liu, L. et al., "LY2875358, a neutralizing and internalizing anti-MET bivalent antibody, inhibits HGF-dependent and HGF-independent MET activation and tumor growth." Clinical Cancer Research 20.23 (2014): 6059-6070.; and Jin, H., "MetMAb, the one-armed 5D5 anti-c-Met antibody, inhibits orthotopic pancreatic tumor growth and improves survival." Cancer Research 68, 11 (2008): 4360-8;

[0154] Antibodies targeting CD70 are known in the art and include, for example, anti-CD70 antibodies described in the following literature: McEarchern, Oflazoglu et al., "Engineered anti-CD70 antibody with multiple effector functions exhibits in vitro and in vivo antitumor activities." Blood 109.3 (2007): 1185-1192. Additional anti-CD70 antibodies include, for example, those described in the following literature: Israel, Gulley et al., "Anti-CD70 antibodies: a potential treatment for EBV+CD70-expressing lymphomas." Molecular cancer therapeutics 4.12 (2005): 2037-2044.

[0155] Antibodies targeting CD44 are known in the art and include, for example, anti-CD44 antibodies described in Wang, Su et al., "CD44 antibody-targeted liposomal nanoparticles for molecular imaging and therapy of hepatocellular carcinoma." Biomaterials 33.20 (2012): 5107-5114. Additional anti-CD44 antibodies include, for example, those described in Kania, Kehat-Stadler and Kupfer, "CD44 antibodies inhibit osteoclast formation." Journal of Bone and Mineral Research 12.8 (1997): 1155-1164.; and Kodama, Toda et al., "Anti-CD44 antibody treatment lowers hyperglycemia and improves insulin resistance, adipose inflammation, and hepatic steatosis in diet-induced obese mice." Diabetes 64.3 (2015): 867-875.

[0156] Antibodies targeting CD123 are known in the art, including, for example, anti-CD123 antibodies, which are described in Lee, Yee et al., "Efficacy of an Fc-modified anti-CD123 antibody (CSL362) combined with chemotherapy in xenograft models of acute myelogenous leukemia in immunodeficient mice." Haematologica 100.7 (2015): 914. Additional anti-CD123 antibodies include, for example, those described in Kovtun, Jones et al., "ACD123-targeting antibody-drug conjugate, IMGN632, designed to eradicate AML while sparing normal bone marrow cells." Blood advances 2.8 (2018): 848-858.

[0157] Antibodies targeting CD37 are known in the art and include, for example, anti-CD37 antibodies described in Oostindie, van der Horst et al., "DuoHexaBody- a novel biparatopic CD37 antibody with enhanced Fc-mediated hexamerization as a potential therapy for B-cell malignancies." Blood cancer journal 10.3(2020):1-13. Additional anti-CD37 antibodies include, for example, those described in the following literature: Deckert, Park et al., "A novel anti-CD37 antibody-drug conjugate with multiple anti-tumor mechanisms for the treatment of B-cell malignancies." Blood, The Journal of the American Society of Hematology 122.20(2013):3500-3510.

[0158] Antibodies targeting CD74 are known in the art and include, for example, the anti-CD74 antibody anti-CD74LL1, which is described in the following literature: Stein, Mattes et al., "CD74: a new candidate target for the immunotherapy of B-cell neoplasms." Clinical Cancer Research 13.18 (2007): 5556s-5563s. Additional anti-CD74 antibodies include, for example, the anti-CD74 antibody anti-CD74LN2, which is described in the following literature: Burton, Ely et al., "CD74 is expressed by multiple myeloma and is a promising target for therapy." Clinical Cancer Research 10.19 (2004): 6606-6611.

[0159] Antibodies targeting IGF1R are known in the art, including, for example, anti-IGF1R antibodies, which are described in Gong, Yao et al., "High expression levels of total IGF-1R and sensitivity of NSCLC cells in vitro to an anti-IGF-1R antibody (R1507)." PloS one 4.10 (2009): e7273. Additional anti-IGF1R antibodies include, for example, anti-IGF1R antibodies, which are described in Cao, Roth et al., "Insulin-like growth factor 1 receptor and response to anti-IGF1R antibody therapy in osteosarcoma." PloS one 9.8 (2014): e106249.

[0160] Antibodies targeting CDH6 are known in the art, including, for example, anti-CDH6 antibodies, which are described in the following literature: Bartolomé, Robles et al., "CDH6-activated αIIbβ3 crosstalks with α2β1 to trigger cellular adhesion and invasion in metastatic ovarian and renal cancers." Molecular Oncology 15.7(2021):1849-1865. Additional anti-CDH6 antibodies include, for example, anti-CDH6 antibodies, which are described in the following literature: Ji, Xu et al., "miR-223-3p inhibits human osteosarcoma metastasisand progression by directly targeting CDH6." Molecular Therapy 26.5(2018):1299-1312.

[0161] Antibodies targeting ROR2 are known in the art, including, for example, anti-ROR2 antibodies described in Morioka, Tanikawa et al., "Orphan receptor tyrosine kinase ROR2 as a potential therapeutic target for osteosarcoma." Cancer science 100.7(2009):1227-1233. Additional anti-ROR2 antibodies include, for example, those described in Goydel, Weber et al., "Affinity maturation, humanization, and co-crystallization of a rabbit anti-human ROR2 monoclonal antibody for therapeutic applications." Journal of Biological Chemistry 295.18(2020):5995-6006.

[0162] Antibodies targeting CD71 are known in the art, including, for example, the anti-CD71 antibody anti-Tfr1H68.4, which is described in Byrne et al., "Ferristatin II promotes degradation of transferrinreceptor-1 in vitro and in vivo." PLoS One 8.7(2013):e70199. Additional anti-CD71 antibodies include, for example, those described in Hamamichi et al., "Novel method for screening functional antibody with comprehensive analysis of its immunoliposome." Scientific reports 11.1(2021):1-13; and Kono et al., "Morphological definition of CD71 positive reticulocytes by various staining techniques and electronmicroscopy compared to reticulocytes detected by an automated hematologyanalyzer." Clinica Chimica Acta 404.2(2009):105-110.

[0163] The antibodies described above are exemplary only and are not meant to limit the scope of the present disclosure in any way.Additional binding agents, including antibodies, suitable for incorporation into the methods and binding agents of the present disclosure will be apparent to those of ordinary skill.

[0164] Although various aspects of the present disclosure have been described with reference to the disclosed embodiments, those skilled in the art will readily appreciate that the specific examples disclosed are merely illustrative of these aspects and in no way limit the present disclosure. Various modifications can be made without departing from the spirit of the present disclosure.

[0165] In some embodiments, the first binding domain comprises a heavy chain (HC) sequence, a variable heavy chain (VH) sequence, a light chain (LC) sequence, and a variable light chain (VL) sequence. In some embodiments, the first binding domain comprises an HC sequence and a VH sequence. The first binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence may comprise one or more sequences listed in Table 1 or Table 4. The first binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence may comprise at least 70% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 75% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 80% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 85% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 90% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 91% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 92% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 93% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 94% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 95% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 96% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 97% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 98% sequence identity to one or more sequences listed in Table 1 or Table 4.In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99.5% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99.9% sequence identity to one or more sequences listed in Table 1 or Table 4.

[0166] In some embodiments, the first binding domain comprises an antibody comprising a heavy chain (HC) sequence, a variable heavy chain (VH) sequence, a light chain (LC) sequence, and a variable light chain (VL) sequence. In some embodiments, the first binding domain comprises an antibody comprising an HC sequence and a VH sequence. The first binding domain comprising an antibody comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence may comprise one or more sequences listed in Table 1 or Table 4. The first binding domain comprising an antibody comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence may comprise at least 70% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an antibody comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 75% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain comprising an antibody comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 80% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 85% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 90% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 91% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 92% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 93% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 94% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 95% sequence identity to one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 96% sequence identity to one or more sequences listed in Table 1 or Table 4.In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 97% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 98% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99.5% sequence identity with one or more sequences listed in Table 1 or Table 4. In some cases, the first binding domain of an antibody comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99.9% sequence identity with one or more sequences listed in Table 1 or Table 4.

[0167] In some embodiments, the first binding domain comprises a sequence listed in Table 1. In some embodiments, the first binding domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.9% sequence identity to a sequence listed in Table 1.

[0168] In some cases, the first binding domain may bind to the same epitope as any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 70% sequence identity to an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 75% sequence identity to an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 80% sequence identity to an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 85% sequence identity to an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 90% sequence identity to an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 95% sequence identity to an epitope bound by any one of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 99% sequence identity to an epitope bound by any one of the antibodies listed in Table 1.

[0169] The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 1 to an epitope that comprises about 70% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 1 to an epitope that comprises about 75% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 1 to an epitope that comprises about 80% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 1 to an epitope that comprises about 85% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 90% sequence identity with an epitope bound by any of the antibodies listed in Table 1 with a similar affinity as any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 95% sequence identity with an epitope bound by any of the antibodies listed in Table 1 with a similar affinity as any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 99% sequence identity with an epitope bound by any of the antibodies listed in Table 1 with a similar affinity as any of the antibodies listed in Table 1.

[0170] In some embodiments, the first binding domain may bind to the same epitope as any of the antibodies listed in Table 1 with a different affinity than any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 70% sequence identity to an epitope bound by any of the antibodies listed in Table 1 with a different affinity than any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 75% sequence identity to an epitope bound by any of the antibodies listed in Table 1 with a different affinity than any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises about 80% sequence identity to an epitope bound by any of the antibodies listed in Table 1 with a different affinity than any of the antibodies listed in Table 1. The first binding domain may bind with a different affinity than any of the antibodies listed in Table 1 to an epitope that comprises about 85% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind with a different affinity than any of the antibodies listed in Table 1 to an epitope that comprises about 90% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind with a different affinity than any of the antibodies listed in Table 1 to an epitope that comprises about 95% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind with a different affinity than any of the antibodies listed in Table 1 to an epitope that comprises about 99% sequence identity with an epitope bound by any of the antibodies listed in Table 1.

[0171] The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope does not bind to any of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any one or more of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any two or more of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any three or more of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any four or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any five or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any six or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any seven or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any eight or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any nine or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any ten or more of the same amino acids on the internalization receptor protein.

[0172] In some embodiments, the antibody targeted to a degrader protein comprises a sequence listed in Table 1. In some embodiments, the antibody targeted to a degrader protein comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.9% sequence identity to the sequence listed in Table 1.

[0173] In some cases, an antibody targeting a degrader protein may bind to the same epitope as any of the antibodies listed in Table 1. An antibody targeting a degrader protein may bind to an epitope that comprises about 70% sequence identity with an epitope bound by any of the antibodies listed in Table 1. An antibody targeting a degrader protein may bind to an epitope that comprises about 75% sequence identity with an epitope bound by any of the antibodies listed in Table 1. An antibody targeting a degrader protein may bind to an epitope that comprises about 80% sequence identity with an epitope bound by any of the antibodies listed in Table 1. An antibody targeting a degrader protein may bind to an epitope that comprises about 85% sequence identity with an epitope bound by any of the antibodies listed in Table 1. An antibody targeting a degrader protein may bind to an epitope that comprises about 90% sequence identity with an epitope bound by any of the antibodies listed in Table 1. The antibody targeting the degrader protein may bind to an epitope that comprises about 95% sequence identity with the epitope bound by any of the antibodies listed in Table 1. The antibody targeting the degrader protein may bind to an epitope that comprises about 99% sequence identity with the epitope bound by any of the antibodies listed in Table 1.

[0174] Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope does not bind to any of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any one or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any two or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any three or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any four or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any five or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any six or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any seven or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any eight or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any nine or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 1, wherein the epitope binds to any ten or more of the same amino acids on the degrader protein.

[0175] Table 1. Exemplary antibody sequences targeting internalizing receptor proteins.

[0176]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196] The sequences listed in Table 1 (SEQ ID NOs: 1-276) are amino acid molecules. The sequences listed in Table 1 (SEQ ID NOs: 1-276) are amino acid molecules as synthetic constructs. The sequences listed in Table 1 for the HC sequence (heavy chain), VH sequence (variable heavy chain sequence), LC sequence (light chain), and VL sequence (variable light chain sequence) (SEQ ID NOs: 1-276) are amino acid molecules as synthetic constructs.

[0197] Table 2. CDR sequences of exemplary antibodies targeting internalizing receptor proteins.

[0198]

[0199]

[0200]

[0201] Table 3. CDR sequences of exemplary antibodies targeting internalizing receptor proteins.

[0202]

[0203]

[0204]

[0205] The sequences listed in Tables 2 and 3 (SEQ ID NOs: 475-885) are amino acid molecules. The sequences listed in Tables 2 or 3 (SEQ ID NOs: 475-885) are amino acid molecules as synthetic constructs. The sequences listed in Tables 2 or 3 for CDRs (complementarity determining regions) (SEQ ID NOs: 475-885) are amino acid molecules as synthetic constructs.

[0206] In some embodiments, the first binding domain comprises at least one complementarity determining region (CDR) sequence. The first binding domain comprising at least one complementarity determining region (CDR) sequence may comprise one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 90% sequence identity with one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising an antibody comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 91% sequence identity with one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 92% sequence identity with one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 93% sequence identity with one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 94% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 95% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 96% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 97% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 98% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99.5% sequence identity to one or more sequences listed in Table 2 or Table 3. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99.99% sequence identity to one or more sequences listed in Table 2 or Table 3.

[0207] In some embodiments, the first binding domain comprises at least one sequence listed in Table 2 or Table 3. In some embodiments, the first binding domain comprises at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9% sequence identity to a sequence listed in Table 2 or Table 3.

[0208] In some embodiments, the first binding domain comprises a sequence listed in Table 4. In some embodiments, the first binding domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.9% sequence identity to a sequence listed in Table 4.

[0209] In some cases, the first binding domain may bind to the same epitope as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 70% sequence identity to an epitope bound by any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 75% sequence identity to an epitope bound by any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 80% sequence identity to an epitope bound by any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 85% sequence identity to an epitope bound by any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 90% sequence identity to an epitope bound by any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 95% sequence identity to an epitope bound by any one of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 99% sequence identity to an epitope bound by any one of the antibodies listed in Table 4.

[0210] In some embodiments, the first binding domain may bind to the same epitope as any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 70% sequence identity to the epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 75% sequence identity to the epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 80% sequence identity to the epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 85% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 90% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 95% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 99% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a similar affinity as any of the antibodies listed in Table 4.

[0211] In some embodiments, the first binding domain may bind to the same epitope as any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 70% sequence identity to an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 75% sequence identity to an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 80% sequence identity to an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 85% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 90% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 95% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises about 99% sequence identity with an epitope bound by any of the antibodies listed in Table 4 with a different affinity than any of the antibodies listed in Table 4.

[0212] The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope does not bind to any of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any one or more of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any two or more of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any three or more of the same amino acids on the internalizing receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any four or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any five or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any six or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any seven or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any eight or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any nine or more of the same amino acids on the internalization receptor protein. The first binding domain may bind to an epitope that comprises a different epitope than the epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any ten or more of the same amino acids on the internalization receptor protein.

[0213] In some embodiments, the antibody targeted to a degrader protein comprises a sequence listed in Table 4. In some embodiments, the antibody targeted to a degrader protein comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.9% sequence identity to the sequence listed in Table 4.

[0214] In some cases, an antibody targeting a degrader protein may bind to the same epitope as any of the antibodies listed in Table 4. An antibody targeting a degrader protein may bind to an epitope that comprises about 70% sequence identity with an epitope bound by any of the antibodies listed in Table 4. An antibody targeting a degrader protein may bind to an epitope that comprises about 75% sequence identity with an epitope bound by any of the antibodies listed in Table 4. An antibody targeting a degrader protein may bind to an epitope that comprises about 80% sequence identity with an epitope bound by any of the antibodies listed in Table 4. An antibody targeting a degrader protein may bind to an epitope that comprises about 85% sequence identity with an epitope bound by any of the antibodies listed in Table 4. An antibody targeting a degrader protein may bind to an epitope that comprises about 90% sequence identity with an epitope bound by any of the antibodies listed in Table 4. The antibody targeting the degrader protein may bind to an epitope that comprises about 95% sequence identity with the epitope bound by any of the antibodies listed in Table 4. The antibody targeting the degrader protein may bind to an epitope that comprises about 99% sequence identity with the epitope bound by any of the antibodies listed in Table 4.

[0215] Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope does not bind to any of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any one or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any two or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any three or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any four or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any five or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any six or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any seven or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any eight or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any nine or more of the same amino acids on the degrader protein. Antibodies targeting degrader proteins may bind to an epitope that comprises a different epitope than an epitope bound by any of the antibodies listed in Table 4, wherein the epitope binds to any ten or more of the same amino acids on the degrader protein.

[0216] Table 4. Additional exemplary antibody sequences targeting degrader proteins.

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225] The sequences listed in Table 4 (SEQ ID NOs: 277-350) are amino acid molecules. The sequences listed in Table 4 (SEQ ID NOs: 277-350) are amino acid molecules as synthetic constructs. The sequences listed in Table 4 for the HC sequence (heavy chain), VH sequence (variable heavy chain sequence), LC sequence (light chain), and VL sequence (variable light chain sequence) (SEQ ID NOs: 277-350) are amino acid molecules as synthetic constructs.

[0226] Table 5. CDR sequences of exemplary antibodies targeting degradation receptor proteins.

[0227]

[0228] Table 6. CDR sequences of exemplary antibodies targeting degradation receptor proteins.

[0229]

[0230]

[0231] The sequences listed in Table 5 or Table 6 (SEQ ID NO: 886-984) are amino acid molecules. The sequences listed in Table 5 or Table 6 (SEQ ID NO: 886-984) are amino acid molecules as synthetic constructs. The sequences listed in Table 5 or Table 6 for CDRs (complementarity determining regions) (SEQ ID NO: 886-984) are amino acid molecules as synthetic constructs.

[0232] In some embodiments, the first binding domain comprises at least one complementarity determining region (CDR) sequence. The first binding domain comprising at least one complementarity determining region (CDR) sequence may comprise one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 90% sequence identity with one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising an antibody comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 91% sequence identity with one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 92% sequence identity with one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 93% sequence identity with one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 94% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 95% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 96% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 97% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 98% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99.5% sequence identity to one or more sequences listed in Table 5 or Table 6. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99.99% sequence identity to one or more sequences listed in Table 5 or Table 6.

[0233] In some embodiments, the first binding domain comprises at least one sequence listed in Table 5 or Table 6. In some embodiments, the first binding domain comprises at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9% sequence identity to a sequence listed in Table 5 or Table 6.

[0234] Second binding region

[0235] In some embodiments, the second binding domain (i.e., cMET binding domain) comprises a cMET binding domain (e.g., CDRs that specifically bind to cMET) derived from an anti-cMET antibody. Such antibodies are known to those skilled in the art and can be incorporated into the methods and binding agents of the present disclosure. Antibodies targeting cMET are known in the art and include, for example, the following anti-cMET antibodies: (i) onatuzumab, which is described in, for example, Lee, D. et al., "Development of antibody-based c-Met inhibitors for targeted cancer therapy." ImmunoTargets and therapy 4 (2015): 35-44; (ii) amivantamab, which is described in, for example, Neijssen, Joost et al., "Discovery of amivantamab (JNJ-61186372), a bispecific antibody targeting EGFR and MET." Journal of Biological Chemistry 296 (2021); (iii) terituzumab, which is described in, for example, Strickler, John H. et al., "Phase I dose-escalation and-expansion study of telisotuzumab (ABT-700), an anti-c-Met antibody, in patients with advanced solid tumors." tumors." Molecular cancer therapeutics 19.5(2020):1210-1217; (iv) REGN5093s58, which is described, for example, in the following literature: Oh, Seung Yeon et al., "Preclinical Study of a Biparatopic METxMET Antibody–Drug Conjugate, REGN5093-M114, Overcomes MET-driven Acquired Resistance to EGFR TKIs in EGFR-mutant NSCLC." Clinical Cancer Research 29.1(2023):221-232; (v) emibetuzumab, also known as LY2875358, which is described, for example, in the following literature: Liu, L.et al., “LY2875358, a aneutralizing and internalizing anti-MET bivalent antibody, inhibits HGF-dependent and HGF-independent MET activation and tumor growth.” Clinical Cancer Research 20.23(2014):6059-6070; (vi) 5D5, which is described, for example, in the following literature: Jin, H., “MetMAb, the one-armed 5D5 anti-c-Met antibody, inhibits orthotopic pancreatic tumor growth and improves survival.” Cancer Research 68,11(2008):4360-8; and (vii) F46, which is described, for example, in the following literature: Young, M., “A new anti-c-Met antibody selected by a mechanism-based dual-screening method: therapeutic potential incancer. Molecules and cells 34,6(2012):523-9.

[0236] The antibodies described above are exemplary only and are not meant to limit the scope of the present disclosure in any way.Additional binding agents, including antibodies, suitable for incorporation into the methods and binding agents of the present disclosure will be apparent to those of ordinary skill.

[0237] In some embodiments, the second binding domain binds to a mutant cMET protein. In some embodiments, the second binding domain selectively binds to a mutant cMET protein.

[0238] In some embodiments, the second binding domain comprises a heavy chain (HC) sequence, a variable heavy chain (VH) sequence, a light chain (LC) sequence, and a variable light chain (VL) sequence. In some embodiments, the second binding domain comprises an HC sequence and a VH sequence. The second binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence may comprise one or more sequences listed in Table 7. The second binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence may comprise at least 70% sequence identity with one or more sequences listed in Table 7. In some cases, the second binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 75% sequence identity with one or more sequences listed in Table 7. In some cases, the second binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 80% sequence identity with one or more sequences listed in Table 7. In some cases, the second binding domain comprising an HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 85% sequence identity with one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 90% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 91% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 92% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 93% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 94% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 95% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 96% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 97% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 98% sequence identity to one or more sequences listed in Table 7.In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99.5% sequence identity to one or more sequences listed in Table 7. In some cases, the second binding domain comprising a HC sequence, a VH sequence, a LC sequence, and a VL sequence comprises at least 99.9% sequence identity to one or more sequences listed in Table 7.

[0239] In some embodiments, the second binding domain comprises a sequence listed in Table 7. In some embodiments, the second binding domain comprises a sequence listed in Table 7. In some embodiments, the second binding domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.9% sequence identity to a sequence listed in Table 7.

[0240] In some embodiments, the second binding domain comprises at least 70% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 75% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 80% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 85% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 90% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 91% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 92% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 93% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 94% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 95% sequence identity with ervantuzumab. In some embodiments, the second binding domain comprises at least 96% sequence identity to ervantuzumab. In some embodiments, the second binding domain comprises at least 97% sequence identity to ervantuzumab. In some embodiments, the second binding domain comprises at least 98% sequence identity to ervantuzumab. In some embodiments, the second binding domain comprises at least 99% sequence identity to ervantuzumab. In some embodiments, the second binding domain comprises at least 99.5% sequence identity to ervantuzumab. In some embodiments, the second binding domain comprises at least 99.9% sequence identity to ervantuzumab.

[0241] In some embodiments, the second binding domain comprises at least 70% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 75% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 80% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 85% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 90% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 91% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 92% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 93% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 94% sequence identity with terituzumab. In some embodiments, the second binding domain comprises at least 95% sequence identity to Terituzumab. In some embodiments, the second binding domain comprises at least 96% sequence identity to Terituzumab. In some embodiments, the second binding domain comprises at least 97% sequence identity to Terituzumab. In some embodiments, the second binding domain comprises at least 98% sequence identity to Terituzumab. In some embodiments, the second binding domain comprises at least 99% sequence identity to Terituzumab. In some embodiments, the second binding domain comprises at least 99.5% sequence identity to Terituzumab. In some embodiments, the second binding domain comprises at least 99.9% sequence identity to Terituzumab.

[0242] In some embodiments, the second binding domain comprises at least 70% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 75% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 80% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 85% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 90% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 91% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 92% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 93% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 94% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 95% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 96% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 97% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 98% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 99% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 99.5% sequence identity with onatumomab. In some embodiments, the second binding domain comprises at least 99.9% sequence identity with onatumomab.

[0243] In some embodiments, the second binding domain comprises at least 70% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 75% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 80% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 85% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 90% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 91% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 92% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 93% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 94% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 95% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 96% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 97% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 98% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 99% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 99.5% sequence identity with REGN5093s58. In some embodiments, the second binding domain comprises at least 99.9% sequence identity with REGN5093s58.

[0244] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 70% sequence identity to an epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by ervantuzumab. In some cases, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 95% sequence identity to an epitope bound by ervantuzumab.

[0245] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope does not comprise any of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one, two, three, four, five, or six of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one or more of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises two or more of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises three or more of the amino acids from the epitope bound by ervantuzumab. In some embodiments, the second binding domain binds an epitope of cMET on a target cell, wherein the epitope comprises four or more of the amino acids from the epitope to which eviantazumab binds.

[0246] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 70% sequence identity to an epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by terituzumab. In some cases, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 95% sequence identity to an epitope bound by terituzumab.

[0247] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope does not comprise any of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one, two, three, four, five, or six of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one or more of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises two or more of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises three or more of the amino acids from the epitope bound by terituzumab. In some embodiments, the second binding domain binds an epitope of cMET on a target cell, wherein the epitope comprises four or more of the amino acids from the epitope bound by terituzumab.

[0248] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 70% sequence identity to the epitope bound by onatumomab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 80% sequence identity to the epitope bound by onatumomab. In some cases, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by onatumomab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 95% sequence identity to the epitope bound by onatumomab.

[0249] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope does not comprise any of the amino acids from the epitope bound by onatumomab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one, two, three, four, five, or six of the amino acids from the epitope bound by onatumomab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one or more of the amino acids from the epitope bound by onatumomab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises two or more of the amino acids from the epitope bound by onatumomab. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises three or more of the amino acids from the epitope bound by onatumomab. In some embodiments, the second binding domain binds an epitope of cMET on a target cell, wherein the epitope comprises four or more of the amino acids from the epitope bound by onatumumab.

[0250] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 70% sequence identity to the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 80% sequence identity to the epitope bound by REGN5093s58. In some cases, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises at least 95% sequence identity to the epitope bound by REGN5093s58.

[0251] In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope does not comprise any of the amino acids from the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one, two, three, four, five, or six of the amino acids from the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises one or more of the amino acids from the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises two or more of the amino acids from the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds to an epitope of cMET on a target cell, wherein the epitope comprises three or more of the amino acids from the epitope bound by REGN5093s58. In some embodiments, the second binding domain binds an epitope of cMET on a target cell, wherein the epitope comprises four or more of the amino acids from the epitope bound by REGN5093s58.

[0252] In some embodiments, the epitope of cMET comprises the following amino acids of human cMET (UniProt ID: P08581): G326, A327, Q328, R331, Q332, 1333, G334, A335, S336, L337, N338, D339, K368, Y369, R426, 1446, G448, D449, and R469. The second binding domain can target an epitope comprising amino acids G326, A327, Q328, R331, Q332, 1333, G334, A335, S336, L337, N338, D339, K368, Y369, R426, 1446, G448, D449, and R469 of human cMET. In some embodiments, an antibody targeting amino acids G326, A327, Q328, R331, Q332, 1333, G334, A335, S336, L337, N338, D339, K368, Y369, R426, 1446, G448, D449, and R469 of human cMET comprises onatumomab. In some embodiments, an epitope of cMET comprises the following amino acids of human cMET: D94, F96, P97, C98, Q99, D100, S103, K104, A105, N106, H159, C160, F162, S163, P164, 1166, E167, T222, and D224. The second binding domain can target an epitope comprising amino acids D94, F96, P97, C98, Q99, D100, S103, K104, A105, N106, H159, C160, F162, S163, P164, I166, E167, T222, and D224 of human cMET. In some embodiments, the antibody targeting amino acids D94, F96, P97, C98, Q99, D100, S103, K104, A105, N106, H159, C160, F162, S163, P164, I166, E167, T222, and D224 of human cMET comprises ivantuzumab.

[0253] In some cases, the second binding domain can bind to the same epitope as entuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain can bind to an epitope that comprises about 70% sequence identity to an epitope bound by entuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain can bind to an epitope that comprises about 75% sequence identity to an epitope bound by entuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain can bind to an epitope that comprises about 80% sequence identity to an epitope bound by entuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain may bind to an epitope that comprises about 85% sequence identity to an epitope bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain may bind to an epitope that comprises about 90% sequence identity to an epitope bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain may bind to an epitope that comprises about 95% sequence identity to an epitope bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain may bind to an epitope that comprises about 99% sequence identity to an epitope bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58.

[0254] The second binding domain may bind to an epitope that comprises a different epitope than that bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58. The second binding domain may bind to an epitope that comprises a different epitope than that bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope does not bind to any of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any one or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by entuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any two or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by entuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any three or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by entuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any four or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by entuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any five or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by entuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any six or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than that bound by entuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any seven or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than the epitope bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any eight or more of the same amino acids on cMET. The second binding domain may bind to an epitope that comprises a different epitope than the epitope bound by ervantuzumab, terituzumab, onatumumab, or REGN5093s58, wherein the epitope binds to any nine or more of the same amino acids on cMET.The second binding domain can bind to an epitope that comprises a different epitope than the epitope bound by ervantuzumab, terituzumab, onatuzumab, or REGN5093s58, wherein the epitope binds to any ten or more of the same amino acids on cMET.

[0255] In some cases, the second binding domain may bind to the same epitope as any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 70% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 75% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 80% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 85% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 90% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 95% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The second binding domain may bind to an epitope that comprises about 99% sequence identity to an epitope bound by any of the antibodies listed in Table 7.

[0256] In some embodiments, the first binding domain may bind to the same epitope bound by any of the antibodies listed in Table 7 with the same affinity as any of the antibodies listed in Table 7. The first binding domain may bind to an epitope that comprises about 70% sequence identity to the epitope bound by any of the antibodies listed in Table 7 with a similar affinity as any of the antibodies listed in Table 7. The first binding domain may bind to an epitope that comprises about 75% sequence identity to the epitope bound by any of the antibodies listed in Table 7 with a similar affinity as any of the antibodies listed in Table 7. The first binding domain may bind to an epitope that comprises about 80% sequence identity to the epitope bound by any of the antibodies listed in Table 7 with a similar affinity as any of the antibodies listed in Table 7. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 7 to an epitope that comprises about 85% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 7 to an epitope that comprises about 90% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 7 to an epitope that comprises about 95% sequence identity to an epitope bound by any of the antibodies listed in Table 7. The first binding domain may bind with a similar affinity as any of the antibodies listed in Table 7 to an epitope that comprises about 99% sequence identity to an epitope bound by any of the antibodies listed in Table 7.

[0257] Table 7. Exemplary cMET-targeting antibody sequences.

[0258]

[0259]

[0260]

[0261]

[0262] The sequences listed in Table 7 (SEQ ID NOs: 351-370) are amino acid molecules. The sequences listed in Table 7 (SEQ ID NOs: 351-370) are amino acid molecules as synthetic constructs. The sequences listed in Table 7 for the HC sequence (heavy chain), VH sequence (variable heavy chain sequence), LC sequence (light chain), and VL sequence (variable light chain sequence) (SEQ ID NOs: 351-370) are amino acid molecules as synthetic constructs.

[0263] Table 8. CDR sequences of exemplary antibodies targeting cMET.

[0264]

[0265]

[0266]

[0267] Table 9. CDR sequences of exemplary antibodies targeting cMET.

[0268]

[0269]

[0270] The sequences listed in Table 8 or Table 9 (SEQ ID NO: 1003-1026) are amino acid molecules. The sequences listed in Table 8 or Table 9 (SEQ ID NO: 1003-1026) are amino acid molecules as synthetic constructs. The sequences listed in Table 8 or Table 9 for CDR (complementarity determining region) sequences (SEQ ID NO: 1003-1026) are amino acid molecules as synthetic constructs.

[0271] In some embodiments, the first binding domain comprises at least one complementarity determining region (CDR) sequence. The first binding domain comprising at least one complementarity determining region (CDR) sequence may comprise one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 90% sequence identity with one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising an antibody comprising an HC sequence, a VH sequence, an LC sequence, and a VL sequence comprises at least 91% sequence identity with one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 92% sequence identity with one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 93% sequence identity with one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 94% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 95% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 96% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 97% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 98% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99.5% sequence identity to one or more sequences listed in Table 8 or Table 9. In some cases, the first binding domain comprising at least one complementarity determining region (CDR) sequence comprises at least 99.99% sequence identity to one or more sequences listed in Table 8 or Table 9.

[0272] In some embodiments, the first binding domain comprises at least one sequence listed in Table 8 or Table 9. In some embodiments, the first binding domain comprises at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9% sequence identity to a sequence listed in Table 8 or Table 9.

[0273] synthesis

[0274] Binding agents are synthesized using recombinant DNA and protein expression techniques. For example, to synthesize DNA encoding the dual IgG disclosed herein, suitable DNA sequences encoding the constant domains of the heavy and light chains are widely available. Sequences encoding the selected variable domains are inserted by standard methods, and the resulting nucleic acids encoding the full-length heavy and light chains are transduced into suitable host cells and expressed. Alternatively, the nucleic acids can be expressed in a cell-free expression system that can provide more control over oxidation and reduction conditions, pH, folding, and glycosylation.

[0275] The binding activity of the engineered antibodies of the present invention can be determined by any suitable method known in the art. For example, the binding activity of the engineered antibodies of the present invention can be determined by, for example, Scatchard analysis (Munsen et al., Analyt Biochem (1980) 107: 220-39). Specific binding can be assessed using techniques known in the art, including but not limited to competitive ELISA, Determination and / or Determination. Antibodies that preferentially bind or specifically bind (used interchangeably herein) to a target antigen or target epitope are terms well known in the art, and methods for determining such specific binding or preferential binding are also known in the art. If an antibody reacts or associates with a specific antigen or epitope more frequently, more quickly, for a longer duration, and / or with greater affinity than when the antibody reacts or associates with an alternative antigen or epitope, the antibody is said to exhibit specific binding or preferential binding. If the antibody binds to the target with greater affinity, avidity, more readily, and / or for a longer duration than when the antibody binds to other substances, the antibody specifically binds or preferentially binds to the target. In addition, if the antibody binds to the target in the sample with greater affinity, avidity, more readily, and / or for a longer duration than when the antibody binds to other substances present in the sample, the antibody specifically binds or preferentially binds to the target. For example, the antibody that specifically binds or preferentially binds to a HER2 epi-position is an antibody that binds to this epi-position with greater affinity, avidity, more easily and / or with longer duration than this antibody in conjunction with other HER2 epi-positions or non-HER2 epi-positions. It can also be understood by reading this definition that, for example, the antibody that specifically binds or preferentially binds to the first target antigen may or may not specifically bind or preferentially bind to the second target antigen. Therefore, specific binding and preferential binding do not necessarily require (although it may include) exclusive binding.

[0276] Nucleic acid molecules

[0277] In one aspect, some embodiments disclosed herein relate to nucleic acid molecules comprising nucleotide sequences encoding the binding agents of the present disclosure, including expression cassettes and expression vectors containing these nucleic acid molecules, which are operably linked to heterologous nucleic acid sequences (such as, for example, regulatory sequences that direct the in vivo expression of the protein in a host cell).

[0278] Also provided herein are vectors, plasmids or viruses containing one or more of the nucleic acid molecules encoding any of the dual binders disclosed herein. The nucleic acid molecules can be contained in vectors that are capable of directing expression of these nucleic acid molecules in cells that have been transformed / transduced with the vector, for example. Suitable vectors for use in eukaryotic and prokaryotic cells are known in the art and are commercially available or readily prepared by a skilled artisan. See, e.g., Sambrook, J., & Russell, DW (2012). Molecular Cloning: A Laboratory Manual (4th ed.). Cold Spring Harbor, NY: Cold Spring Harbor Laboratory and Sambrook, J., & Russell, DW (2001). Molecular Cloning: A Laboratory Manual (3rd ed.). Cold Spring Harbor, NY: Cold Spring Harbor Laboratory (collectively referred to herein as "Sambrook"); Ausubel, FM (1987). Current Protocols in Molecular Biology. New York, NY: Wiley (including supplements through 2014); Bollag, DM et al., (1996). Protein Methods. New York, NY: Wiley-Liss; Huang, L. et al., (2005). Nonviral Vectors for Gene Therapy. San Diego: Academic Press; Kaplitt, MG et al., (1995). Viral Vectors: Gene Therapy and Neuroscience Applications. San Diego, CA: Academic Press; Lefkovits, I. (1997). The Immunology Methods Manual: The Comprehensive Sourcebook of Techniques. San Diego, CA: Academic Press; Doyle, A. et al., (1998). Cell and Tissue Culture: Laboratory Procedures in Biotechnology. New York, NY: Wiley; Mullis, KB, Ferré, F. & Gibbs, R. (1994).PCR: The Polymerase Chain Reaction. Boston: Birkhauser Publisher; Greenfield, EA (2014). Antibodies: A Laboratory Manual (2nd Edition). New York, NY: Cold Spring Harbor Laboratory Press; Beaucage, SL et al., (2000). Current Protocols in Nucleic Acid Chemistry. New York, NY: Wiley (including supplements through 2014); and Makrides, SC (2003). Gene Transfer and Expression in Mammalian Cells. Amsterdam, NL: Elsevier Sciences BV, the disclosures of which are incorporated herein by reference.

[0279] Binding method on target cancer cells

[0280] In some embodiments, a binding agent comprising a first binding domain that specifically binds to membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell. In some embodiments, a binding agent comprising a first binding domain that specifically binds to membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a gastric adenocarcinoma cancer cell. In some embodiments, a binding agent comprising a first binding domain that specifically binds to membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a non-small cell lung cancer cell. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cell selected from the group consisting of breast cancer cells, B cell lymphoma cells, pancreatic cancer cells, Hodgkin lymphoma cells, ovarian cancer cells, prostate cancer cells, mesothelioma cells, lung cancer cells, non-Hodgkin B cells (B-NHL) cells, melanoma cells, chronic lymphocytic leukemia cells, acute lymphocytic leukemia cells, neuroblastoma cells, glioma cells, glioblastoma cells, bladder cancer cells, colorectal cancer cells, and head and neck cancer cells. In some embodiments, the cancer cell comprises a genetic mutation selected from a cMET exon 14 skipping mutation or a cMET duplication mutation.

[0281] In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell and reduces the expression of cMET on the cancer cell by at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, or at least 80%. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell and reduces the expression of cMET on the cancer cell by about 40% to 80%, about 50% to 80%, about 60% to 80%, about 70% to 80%, about 40% to 70%, about 50% to 70%, about 60% to 70%, about 40% to 60%, or about 50% to 60%. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the expression of cMET in the cancer cell following contact with the binding agent is at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, or at least 80% less than the expression of cMET in a control cancer cell contacted with the monospecific cMET binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the expression of cMET in the cancer cell following contact with the binding agent is at least 40% to 80%, about 50% to 80%, about 60% to 80%, about 70% to 80%, about 40% to 70%, about 50% to 70%, about 60% to 70%, about 40% to 60%, or about 50% to 60% less than the expression of cMET in a control cancer cell contacted with the monospecific cMET binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the expression of cMET in the cancer cell after contact with the binding agent is at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, or at least 80% less than the expression of cMET in a control cancer cell not contacted with the binding agent.In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the expression of cMET in the cancer cell after contact with the binding agent is at least 40% to 80%, about 50% to 80%, about 60% to 80%, about 70% to 80%, about 40% to 70%, about 50% to 70%, about 60% to 70%, about 40% to 60%, or about 50% to 60% less than the expression of cMET in a control cancer cell not contacted with the binding agent.

[0282] In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell and increases surface removal of cMET from the target cancer cell by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell increases cell surface removal of cMET by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%. In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET on the surface of the cancer cell after contact with the binding agent is less than the amount of cMET on the surface of a control cancer cell that has not been contacted with the binding agent by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%. In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET on a target cancer cell, and the amount of cMET on the surface of the cancer cell after contact with the binding agent is less than the amount of cMET on the surface of a control cancer cell that has not been contacted with the binding agent by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 80%, about 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%.In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET on the surface of the cancer cell after contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% less than the amount of cMET on the surface of a control cancer cell contacted with the monospecific cMET binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET on a target cancer cell, and the amount of cMET on the surface of the cancer cell after contact with the binding agent is less than the amount of cMET on the surface of a control cancer cell contacted with the monospecific cMET binding agent by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about %, about 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%.

[0283] In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell and increases the internalization of cMET on the target cancer cell by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell increases the internalization of cMET by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 20-80%, about 30-80%, about 40 In some embodiments, internalization of cMET on target cells is determined relative to internalization of cMET on control cancer cells that have not been contacted with the binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET internalized by the cancer cell after contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% greater than the amount of cMET internalized by a control cancer cell not contacted with the binding agent.In some embodiments, the binding agent comprises a first binding domain that specifically binds to a membrane-associated internalization or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell, and the amount of cMET internalized by the cancer cell after contact with the binding agent is greater than the amount of cMET internalized by a control cancer cell not contacted with the binding agent by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 100-100%, about 150-10 ... In some embodiments, internalization of cMET on target cells is determined relative to internalization of cMET on control cancer cells that have not been contacted with the binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET internalized by the cancer cell following contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% greater than the amount of cMET internalized by a control cancer cell contacted with the monospecific cMET binding agent. In some embodiments, the binding agent comprises a first binding domain that specifically binds to a membrane-associated internalization or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell, and the amount of cMET internalized by the cancer cell after contact with the binding agent is about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, or more than the amount of cMET internalized by a control cancer cell contacted with the monospecific cMET binding agent. %, about 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%.

[0284] In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell and increases the degradation of cMET on the target cancer cell by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell increases the degradation of cMET by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 20-80%, about 30-80%, about 40 In some embodiments, the degradation of cMET on target cells is determined relative to the degradation of cMET on control cancer cells that have not been contacted with the binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET degraded in the cancer cell after contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% greater than the amount of cMET degraded in a control cancer cell not contacted with the binding agent.In some embodiments, the binding agent comprises a first binding domain that specifically binds to membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell and the amount of cMET degraded in the cancer cell after contact with the binding agent is about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 100-100%, about 150-10 ... In some embodiments, the degradation of cMET on target cells is determined relative to the degradation of cMET on control cancer cells that have not been contacted with the binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET degraded in the cancer cell after contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% greater than the amount of cMET degraded in a control cancer cell contacted with the monospecific cMET binding agent. In some embodiments, the binding agent comprises a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell, and the amount of cMET degraded in the cancer cell following contact with the binding agent is about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, or more than the amount of cMET degraded in a control cancer cell contacted with the monospecific cMET binding agent. 0%, about 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%.

[0285] In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET dimer on the cancer cell after contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% less than the amount of degraded cMET in a control cancer cell not contacted with the binding agent. In some embodiments, the binding agent comprises a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET on the target cancer cell, and the amount of cMET dimer on the cancer cell after contact with the binding agent is less than the amount of degraded cMET in a control cancer cell not contacted with the binding agent by about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, or about 90%. About 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET dimer on the cancer cell after contact with the binding agent is at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% less than the amount of degraded cMET in a control cancer cell contacted with the monospecific cMET binding agent.In some embodiments, the binding agent comprises a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET on a target cancer cell, and the amount of cMET dimer in the cancer cell following contact with the binding agent is about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, or about 90-100% less than the amount of cMET degraded in a control cancer cell contacted with the monospecific cMET binding agent. 0%, about 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or about 20-30%.

[0286] In some embodiments, a binding agent comprising a first binding domain that specifically binds to membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell, and after contact with the binding agent, the amount of cMET activated in the cancer cell is less than the amount of cMET degraded in a control cancer cell that has not been contacted with the binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds to membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell, and after contact with the binding agent, the amount of cMET activated in the cancer cell is greater than the amount of cMET degraded in a control cancer cell that has not been contacted with the binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET activated in the cancer cell following contact with the binding agent is within 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 150%, 200%, or 300% of the amount of cMET degraded in a control cancer cell not contacted with the binding agent. In some embodiments, the binding agent comprises a first binding domain that specifically binds to a membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds to cMET on a target cancer cell, and the amount of cMET activation in the cancer cell after contact with the binding agent is about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 2 ... 0-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40%, or within about 20-30%.

[0287] In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET activated in the cancer cell following contact with the binding agent is less than the amount of cMET degraded in a control cancer cell contacted with the monospecific binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell, and the amount of cMET activated in the cancer cell following contact with the binding agent is greater than the amount of cMET degraded in a control cancer cell contacted with the monospecific binding agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds membrane-associated internalization or degradation protein and a second binding domain that specifically binds cMET is contacted with a target cancer cell and the amount of cMET activated in the cancer cell following contact with the binding agent is within 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, 150%, 200%, or 300% of the amount of cMET degraded in a control cancer cell contacted with the monospecific binding agent. In some embodiments, the binding agent comprises a first binding domain that specifically binds membrane-associated internalization protein or degradation protein and a second binding domain that specifically binds cMET on a target cancer cell, and the amount of cMET activation in the cancer cell following contact with the binding agent is about 20-90%, about 30-90%, about 40-90%, about 50-90%, about 60-90%, about 70-90%, about 80-90%, about 100-100%, about 150-100%, about 160-170%, about 170-180%, about 180-190%, about 190-200%, about 200-210%, about 210-220%, about 220-230%, about 230-240%, about 240-250%, about 250-260%, about 260-270%, about 270-280%, about 280-300%, about 290-310%, about 30 ... 20-80%, about 30-80%, about 40-80%, about 50-80%, about 60-80%, about 70-80%, about 20-70%, about 30-70%, about 40-70%, about 50-70%, about 60-70%, about 20-60%, about 30-60%, about 40-60%, about 50-60%, about 20-50%, about 30-50%, about 40-50%, about 20-40%, about 30-40% or within about 20-30%.

[0288] In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or a degradation protein and a second binding domain that specifically binds to cMET contacts a target cancer cell and increases the sensitivity of the cancer cell to a cancer therapeutic agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or a degradation protein and a second binding domain that specifically binds to cMET contacts a target cancer cell and increases the sensitivity of the cancer cell to a cytotoxic agent. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or a degradation protein and a second binding domain that specifically binds to cMET contacts a target cancer cell and reduces the proliferation of the target cancer cell. In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization protein or a degradation protein and a second binding domain that specifically binds to cMET contacts a target cancer cell and increases the death of the cancer cell.

[0289] In some embodiments, a binding agent comprising a first binding domain that specifically binds to a membrane-associated internalization or degradation protein and a second binding domain that specifically binds to cMET is contacted with a target cancer cell in vivo.

[0290] Pharmaceutical composition

[0291] In some embodiments, the binding agents, nucleic acids, and recombinant cells of the present disclosure can be incorporated into compositions (including pharmaceutical compositions). Such compositions typically comprise a binding agent and a pharmaceutically acceptable excipient (e.g., a carrier). The binding agents of the present disclosure can be administered using formulations for administering antibodies and antibody-based therapeutics, or using formulations based thereon.

[0292] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.

[0293] Administration of binding agents

[0294] Administration of any one or more of the therapeutic compositions described herein (eg, binding agents and pharmaceutical compositions) can be used to treat an individual suffering from a neoplastic disease, such as cancer.

[0295] Thus, in one aspect, provided herein are methods for inhibiting target cell activity in an individual, the methods comprising administering to the individual a first therapy comprising one or more of the binding agents and pharmaceutical compositions provided herein, wherein the first therapy inhibits the activity of the target cell by degrading a target surface protein. For example, the activity of the target cell can be inhibited by reducing the proliferation of the target cell, by reducing the pathological or pathogenic behavior of the target cell, by destroying or killing the target cell, etc. In general, the target cell of the disclosed methods can be any cancer cell.

[0296] In some embodiments, a method for treating cancer in a subject comprises administering to the subject a binding agent, wherein the binding agent comprises a first binding domain that specifically binds a membrane-associated internalization or degradation protein and a second binding domain that specifically binds a target protein, wherein the membrane-associated internalization or degradation protein is expressed on a target cell, wherein the target protein comprises cMET.

[0297] In some embodiments, the binding agents disclosed herein can be compared with other binding agents. In some cases, other binding agents are monospecific binding agents. In some embodiments, the monospecific binding agent is terituzumab. In some embodiments, the monospecific binding agent is onatuzumab. In some embodiments, the monospecific binding agent is REGN5093s58. In some cases, other binding agents can target membrane-associated proteins of non-cMET. In some cases, other binding agents can target degraded proteins of non-cMET. In some cases, other binding agents can bind to RSV F protein. In some cases, other binding agents may not bind to the target. In some embodiments, the binding domain configured to bind to a control (e.g., RSV) includes the sequences listed in Table 10. In some embodiments, the binding domain configured to bind to a control (e.g., RSV) includes the sequences listed in Table 11. In some cases, other binding agents are monospecific binding agents. In some embodiments, the monospecific binding agent includes the sequences listed in Table 12.

[0298] Table 10. Exemplary Binder Control Arms for Arm 1

[0299]

[0300]

[0301]

[0302]

[0303] Table 11. Exemplary Binder Control Arms for Arm 2

[0304]

[0305]

[0306]

[0307] Table 12. Exemplary monospecific antibody controls

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315] The sequences listed in Tables 10, 11, and 12 (SEQ ID NOs: 371-471) are amino acid molecules. The sequences listed in Tables 10, 11, and 12 (SEQ ID NOs: 371-471) are amino acid molecules as synthetic constructs. The sequences listed in Tables 10, 11, and 12 for the HC sequence (heavy chain), VH sequence (variable heavy chain sequence), LC sequence (light chain), and VL sequence (variable light chain sequence) (SEQ ID NOs: 371-471) are amino acid molecules as synthetic constructs.

[0316] Example

[0317] The following examples are illustrative and non-limiting of the scope of the compositions, devices, and methods disclosed herein.

[0318] Cell lines:

[0319] Cells were grown in complete growth medium and maintained at 37°C and 5% CO2.

[0320] Example 1 - Bispecific Antibody Expression:

[0321] According to the manufacturer's protocol, transient transfection was used to express and purify bispecific antibodies from mammalian cells (exemplary: Expi293F, ExpiCHO-S). At designated time points (exemplary: 4-14 days), the culture medium was harvested by centrifugation at 4,000 x g for 20 min. The tagged bispecific antibodies and pestle half IgG were purified by Ni-NTA or protein A affinity chromatography, and the buffer was exchanged into PBS containing 20% ​​glycerol, concentrated, and quickly frozen for storage at -80°C. IgG and hole structure half IgG were purified by protein A affinity chromatography, and the buffer was exchanged into PBS containing 20% ​​glycerol. Knob-in-hole half IgG was reconstituted under reducing conditions (exemplary: 10 mM Tris pH 7.5, 100 mM NaCl, 20% 800 mM L-Arg pH 10 plus 200-fold excess reduced glutathione) and then purified by Ni-NTA affinity chromatography, buffer exchanged to PBS containing 20% ​​glycerol, concentrated, and flash frozen for storage at -80° C. The purity and integrity of all proteins were assessed by SDS-PAGE and SEC.

[0322] Example 2 - Stable cell line generation:

[0323] Receptors (e.g., cMET) with N-terminal epitope tags (exemplary: alfa, HA, Myc, etc.) were cloned into the pLVX lentiviral vector. Lentivirus was produced by transfecting HEK293T cells with standard packaging vectors. Stable cell lines expressing epitope-tagged receptors were selected with puromycin, and expression was verified by flow cytometry using an anti-epitope tag primary antibody.

[0324] Example 3-degradation experiment:

[0325] Cells (illustrative examples: human cancer cell lines, primary human immune cells, or stable cell lines produced herein) are plated (illustrative examples: in 6, 12, 24, 48, 96, or 348-well plates) and grown to ~70% confluence before treatment. The culture medium is aspirated, and the cells are treated with bispecific antibodies (including, for example, any antibody disclosed herein) or control antibodies in complete growth medium (concentration range: 0.001 to 1000 nM; time range: 0-7 days). After incubation at 37°C, the cells are washed with phosphate-buffered saline (PBS). The samples are then tested according to Western blotting, intracellular Western blotting, or flow cytometry protocols to quantify target protein levels.

[0326] Example 4 - Quantification of cMET levels by Western blotting:

[0327] Cells were lifted with versene and harvested by centrifugation at 300xg for 5 min at 4°C. The cell pellet was lysed at 4°C with 1x RIPA buffer (Sigma-Aldrich) containing cOmplete mini protease inhibitor cocktail for 30 min. The lysate was centrifuged at 2,000 (for 96-well plates) or 16,000xg for 10 min at 4°C. 4x NuPAGE LDS sample buffer (Invitrogen) and 2-mercaptoethanol (BME) were added to the lysate and boiled for 10 min. Equal amounts of lysate were loaded onto a 4-12% Bis-Tris gel and run at 200V for 37 min. The gel was incubated in 20% ethanol for 10 min and transferred to a polyvinylidene fluoride (PVDF) membrane. The membrane was blocked in PBS containing 0.1% Tween-20 + 5% bovine serum albumin (BSA) at room temperature with gentle shaking for 30 min. The membrane was incubated with the corresponding dilution of the primary antibody in PBS + 0.2% Tween-20 + 5% BSA at room temperature for 1 hour under gentle shaking. The membrane was washed four times with tris-buffered saline (TBS) + 0.1% Tween-20 and then incubated with the secondary antibody in PBS + 0.2% Tween-20 + 5% BSA at room temperature for 1 hour. The membrane was washed four times with TBS + 0.1% Tween-20 and then washed with PBS. The membrane was imaged using an Odyssey CLx imager (LI-COR). Band intensities were quantified using Image Studio software (LI-COR).

[0328] Example 5 - Quantification of cMET levels by in-cell Western blotting

[0329] Fixation solution (exemplary: 4% paraformaldehyde in PBS) is added to the cells and incubated at room temperature for 20min without stirring. The fixation solution is then removed and the cells are washed with PBS. Permeabilization solution (exemplary: 0.1% Triton-X100 in PBS) is added to the cells and incubated under vibration for 20min. The permeabilization solution is removed and the cells are incubated at room temperature under vibration for 1 hour in blocking buffer. The blocking buffer is removed and the cells are incubated with a primary antibody for 2 hours under vibration. The cells are washed four times with TBS+0.1% Tween-20. The cells are then incubated with a secondary antibody for 1 hour under vibration. The cells are then washed four times with TBS+0.1% Tween-20. The washing solution is removed and the plate is imaged using an Odyssey CLx imager (LI-COR). Empiria Studo software (LI-COR) is used to quantify the pore intensity.

[0330] Example 6 - Quantification of cMET levels by flow cytometry

[0331] Cells were raised with versene and harvested by centrifugation at 300xg for 5min at 4°C. The cell pellet was washed with cold PBS and centrifuged at 300xg for 5min. The cells were blocked with cold PBS+3% BSA and centrifuged (300xg, 5min). The cells were incubated at 4°C for 30min with the primary antibody diluted in PBS+3% BSA. The cells were washed three times with cold PBS+3% BSA, and the secondary antibody (if applicable) diluted in PBS+3% BSA was added and incubated at 4°C for 30min. The cells were washed three times with cold PBS+3% BSA and resuspended in cold PBS. Flow cytometry was performed on a CytoFLEX flow cytometer (Beckman Coulter) and single cells and living cells were gated, and 10,000 cells were obtained. Analyzed using the FlowJo software package.

[0332] Example 7 - Cell surface removal of cMET using bispecific antibodies that bind cMET and a degrader protein.

[0333] To determine cMET cell surface removal by bispecific antibodies that bind to cMET and a degrader protein (bispecific antibodies), the cMETxCD71 bispecific antibody (antibody that binds to cMET and CD71; Figure 1 A- Figure 2D Cell surface depletion assays were performed using the cMET-targeting bispecific antibodies (Table 13). The cMET-targeting bispecific antibodies had terituzumab, onatumumab, ervantuzumab, or REGN5093s58 (four published beta-MET binders that bind to different epitopes on the extracellular portion of cMET) as the cMET-binding domain and ABBV2029 as the CD71-binding arm.

[0334] In addition, an IgG1 isotype control (RG196-1) and palivizumab IgG against RSV (EPI692-1) were used as non-targeting controls. Palivizumab / terituzumab (EPI1086-1), palivizumab / onatuzumab (EPI1087-1), palivizumab / ervantuzumab (EPI1088-1), and palivizumab / Regeneron Seq58 (EPI1098-1) (RSV x cMET bispecific antibodies) were tested as single-arm cMET binding controls, where the second arm does not bind to target cells. Evantuzumab / zalutumumab (EPI818-1; cMET x EGFR) was used as an additional control.

[0335] Table 13. Antibodies tested in the cell surface depletion assay

[0336]

[0337]

[0338] The various constructs were expressed in the gastric adenocarcinoma cell line Hs746T ( Figure 2A ) and non-small cell lung cancer cell lines using multiple different cMET isoforms, NCI-H1993 ( Figure 2B )、NCI-H1975( Figure 2C ) and NCI-H596( Figure 2D ) were tested at a concentration of 50 nM (Table 14). In these assays, HS746T, NCI-H1993, HCI-1975 or NCI-H586 cells were seeded in 96-well plates and incubated overnight at 37°C and 5% CO2. The next morning, the cells were treated with 50 nm or 500 nM of the test antibody. After 24 hours of treatment, the cells were harvested using a dissociation reagent and stained with a fluorescently labeled anti-cMET antibody. The fluorescence intensity of the cells was measured on a Cytek Northern Lights flow cytometer. After considering the background of the isotype control, the percentage of cMET cell surface removal was calculated as the relative difference between the amount of cMET on the cell surface using the test antibody and the amount of cMET on the cell surface using the untreated control sample.

[0339] Table 14. Cancer cell lines tested in the cell surface removal assay

[0340]

[0341] In Hs746T cells ( Figure 2A ), NCI-H1993 cells ( Figure 2B ), NCI-H1975 cells ( Figure 2C ) and NCI-H596 cells ( Figure 2D), all four cMET x CD71 bispecific antibodies induced the removal of cMET from the cell surface. In multiple cell lines (Hs746T, NCI-H1993, NCI-H1975, NCI-H596), the cMET x CD71 bispecific antibodies had higher levels of cMET cell surface removal than entuzumab (a standard of care molecule) and palivizumab. For almost all corresponding pairs, the cMET x CD71 bispecific antibodies also had higher levels of cMET cell surface removal than the single-arm control. This effect was persistent across multiple epitopes because the four cMET binders bound to different epitopes on the extracellular portion of cMET. It was also persistent across a range of cMET mutation states, including MET amplification, MET normal, and METΔex14. cMET genomics is variable in lung cancer. Several genomic alterations are now well defined in clinical samples. The Met exon 14 skipping (METΔex14) mutation causes "skipping" of exon 14 by disrupting the splice site. There is also cMET "amplification," or multiple copies of the MET gene, present in the genome. Based on these results, entuzumab was selected as the cMET-binding arm of the bispecific antibody in additional testing because it had excellent activity in all cell lines tested and low baseline activity in single-arm controls.

[0342] This data demonstrates the effectiveness of bispecific antibodies that bind both cMET and a degrader protein in removing cMET from the surface of target cells under multiple circumstances.

[0343] Example 8 - cMET cell surface depletion screen to identify potent degrader protein binding domains of cMET-targeting bispecific antibodies.

[0344] To identify the degrader protein binding domains on cMET-targeting bispecific antibodies that lead to high cMET cell surface removal, a screen was performed using 78 bispecific antibodies ( Figure 3A-Figure 3B ). 78 bispecific antibodies bind to 18 unique degrader proteins. For most degrader proteins, multiple binding domains binding to different epitopes were tested. In addition, entuzumab (EPI818), emtansine (EPI1444), IgG1 isotype control (RG196-1), and cMET x RSV (EPI1088) were tested as SoC control, cMET only control, negative control, and single-arm control, respectively, with cMET x RSV used as a baseline for comparison. The first binding arm of the bispecific antibody in this embodiment is listed in Table 1 or Table 4. The second binding arm of the bispecific antibody in this embodiment is listed in Table 7. The control arm and antibody are also listed in Table 10, Table 11, and Table 12.

[0345] Screening was performed on the NCI-H1975 cell line (non-small cell lung cancer) using the method previously described in Example 7. Briefly, cells were seeded in 96-well plates and incubated overnight at 37°C and 5% CO2. The next morning, the cells were treated with 50nM of the bispecific antibody or control antibody. After 24 hours of treatment, the cells were harvested using a dissociation reagent and stained with a fluorescently labeled anti-cMET antibody. The fluorescence intensity of the cells was measured on a Cytek Northern Lights flow cytometer. After accounting for the background of the isotype control, the percentage of cMET cell surface removal was calculated as the relative difference between the amount of cMET on the cell surface using the test antibody and the amount of cMET on the cell surface using the untreated control sample.

[0346] The screening identified a panel of degrader proteins and specific molecular epitopes that, when paired with cMET in a bispecific antibody format, demonstrated improved ability to induce cMET cell surface removal compared to palivizumab x cMET (a bispecific antibody pairing cMET with a non-targeting control arm). Figure 3A and Figure 3B ).

[0347] To extend the applicability of the findings from initial screening efforts to more functionally relevant cMET models, the cMET cell surface depletion assay was repeated for priority molecules in cell lines with MET exon 14 deletions and / or MET copy number amplifications frequently observed in patients ( Figures 4A-4C ). The top hits showed strong cMET cell surface removal activity in all cell lines tested. In addition, these experiments demonstrated the cell specificity of cMET degradation using various bispecific antibody pairs.

[0348] Example 9 - cMET internalization screen to identify potent degrader protein binding domains of cMET-targeted bispecific antibodies:

[0349] To further screen and validate the potent degrader protein binding domains identified in the cell surface removal screen, a cMET internalization assay was performed using 20 bispecific antibodies ( Figure 5). 19 bispecific antibodies bind cMET (as a target protein) and 5 unique degrader proteins. For previously identified degrader proteins, multiple binding domains that bind to different epitopes were tested. Enantuzumab (EPI818), imatinib (EPI1444), IgG1 isotype control (RG196) and cMET x RSV (EPI1088) were tested as SoC controls, cMET controls only, negative controls and single-arm controls, respectively, with cMET x RSV used as a baseline for comparison. In addition, for each bispecific antibody, a single-arm control was tested, which contained the same degrader protein binding arm and RSV binding arm as the bispecific antibody. The first binding arm of the bispecific antibody in this embodiment is listed in Table 1 or Table 4. The second binding arm of the bispecific antibody in this embodiment is listed in Table 7. The control arm and antibody are also listed in Table 10, Table 11 and Table 12.

[0350] For internalization assays, NCI-H1975 cells were plated at 7 × 10 3 Cells were plated at a density of 10 cells / well in a 96-well clear culture plate. After culturing for approximately 16 hours, the test antibody was mixed with the rehydrated pH antibody labeling reagent at a molar ratio of 1:3 test antibody to antibody labeling reagent at 37°C for 15 minutes. The labeled antibody was dispensed onto the cells at a concentration of 50 nM. The plate was placed on Images were acquired in a live cell analysis system. Internalization images were sampled at 0 minutes and at 45-minute intervals over a 72-hour period. Image analysis was performed using Incucyte's basic analysis software. Background was subtracted using the "Top-Hat" background subtraction method to obtain the percentage of "Red Object Intensity."

[0351] The screening results identified a panel of degrader proteins and specific molecular epitopes that, when paired with cMET in a bispecific antibody format, demonstrated improved cMET internalization compared to cMET x RSV (a bispecific antibody that pairs cMET with a non-targeting control arm). Many bispecific antibodies showed high internalization. Several of these hits (MUC1, CDH3, and ITGB6) showed synergistic internalization activity, two of which were also identified in the cell surface removal assay. This demonstrates the consistency of the assay and that cMET bispecific antibodies can cause synergistic internalization through identifiable degrader protein binding domains.

[0352] Example 10 - Whole-cell degradation screen to identify potent degrader protein-binding domains of cMET-targeting bispecific antibodies.

[0353] To measure degradation of the target protein, whole-cell degradation of cMET was tested using an AlphaLISA assay and Western blotting.

[0354] For AlphaLISA, 12 bispecific antibodies binding to 6 unique degrader proteins were screened using this assay ( Figures 6A-6C ). For previously identified degrader proteins, multiple binding domains binding to different epitopes were tested. Evantuzumab (EPI818-2), imatinib (EPI1444-1), IgG1 isotype control (RG196-1), and cMET xRSV (EPI1088-2; EPI2132) were tested as SoC controls, cMET controls only, negative controls, and single-arm controls, respectively, with cMET x RSV used as a baseline for comparison. The first binding arm of the bispecific antibody in this embodiment is listed in Table 1 or Table 4. The second binding arm of the bispecific antibody in this embodiment is listed in Table 7. The control arm and antibody are also listed in Table 10, Table 11, and Table 12.

[0355] In the AlphaLISA assay, NCI-H1975 cells, NCI-H596 cells, or Hs746T cells were seeded in reduced serum medium in 384-well plates. After approximately 16 hours of culture, a single concentration of antibody was added to the cells in reduced serum medium and treated for 48 hours. The medium was removed and the cells were lysed. AlphaLISA acceptor beads and biotinylated antibody were added to the lysate and incubated at room temperature for 1 hour. AlphaLISA donor beads were added to the lysate and incubated at room temperature for 2 hours. The plates were read on a Perkin Elmer Envision to determine total cMET levels.

[0356] AlphaLISA screening identified a panel of degrader proteins and specific molecular epitopes that, when paired with cMET in a bispecific antibody format, demonstrated improved whole-cell degradation of cMET compared to palivizumab x cMET (a bispecific antibody pairing cMET with a non-targeting control arm). Degrader proteins identified as potent inducing cMET degradation included CD71, MUC1, CD276, CDH3, TROP2, and EpCAM. Results also demonstrated that cMET degradation was more robust in NCI-H1975 cells than in Ns746T cells.

[0357] In addition, whole-cell degradation of nine bispecific antibodies binding to five different degrader proteins was assessed using Western blotting ( Figures 7A-7C). Enantuzumab (EPI818-2), imatinib (EPI1444-1), IgG1 isotype control (RG196-1), and cMET x RSV (EPI1088-2; EPI2132) were tested as SoC control, cMET only control, negative control, and single-arm control, respectively, with cMET x RSV used as a baseline for comparison. The first binding arm of the bispecific antibody in this example is listed in Table 1 or Table 4. The second binding arm of the bispecific antibody in this example is listed in Table 7. Control arms and antibodies are also listed in Tables 10, 11, and 12.

[0358] For Western blotting, NCI-H1975 cells were seeded at a density of 4e5 cells in 6-well tissue culture plates. After approximately 16 hours of culture, antibodies were added to cells in serum-starved medium at concentrations of 5, 50, or 500 nM and treated for 48 hours. The medium was removed and the cells were lysed. The prepared samples were loaded onto 4-12% BisTris gels and transferred to PVDF membranes. The membranes were probed with cMET or the housekeeping gene β-actin ( Figure 7A-7B Data were quantified using Empiria studio and percentage degradation was normalized to β-actin and compared to PBS control ( Figure 7C By Western blotting, many bispecific antibodies showed improved whole-cell degradation of cMET compared to cMET x RSV (a bispecific antibody that pairs cMET with a non-targeting control arm). Specifically, degrader proteins identified as effective in inducing cMET degradation included MUC1, CD276, CDH3, and TROP2, validating the AlphaLISA results. To further validate the AlphaLISA results, the experiment was repeated using Hs746t cells, and no significant cMET degradation was obtained ( Figure 9 ).

[0359] Subsequently, the potent bispecific antibodies were tested for their ability to inhibit signaling events in NCI-H1975 cells. This was performed by measuring the ratio of pERK to ERK using the Western blot method described above ( Figures 8A-8C The results showed that many effective bispecific antibodies caused a decrease in pERK levels, indicating that bispecific binding agent-induced degradation can inhibit signaling events downstream of the degradation target.

[0360] Example 11 - Reduced dimerization of cMET-targeting bispecific antibodies.

[0361] Current clinical cMET antibodies dimerize cMET and activate downstream signaling associated with oncogenicity. To determine the amount of cMET dimerization that occurs on the cell surface due to the bispecific antibody, a dimerization assay was performed ( Figures 10A-10D ). Enantuzumab (cMET x EGFR; EPI445) or monospecific antibodies specific for cMET (onatoxin, EPI444; terituzumab, EPI443) were compared with bispecific cMET x RSV antibodies (EPI2150; EPI2153; EPI2132). Palivizumab hIgG1 (RSV) was used as an additional control. The first binding arm of the bispecific antibodies in this example is listed in Table 1 or Table 4. The second binding arm of the bispecific antibodies in this example is listed in Table 7. Control arms and antibodies are also listed in Tables 10, 11, and 12.

[0362] In the dimerization assay, the U2OSc-MET / c-MET dimerization cell line. This cell line uses enzyme fragment complementation (EFC) technology, in which β-galactosidase (β-gal) is divided into two fragments (named ProLink and EnzymeAcceptor) to detect ligand-induced c-MET receptor dimerization. These cells are engineered to overexpress c-MET fused to ProLink and c-MET fused to EnzymeAcceptor. After c-MET dimerization, forced complementation of these fragments produces functional β-gal, which can be used to generate a chemiluminescent signal after adding substrate. The cells are cultured in 96-well plates. Antibodies or hepatocyte growth factor (HGF is the natural ligand of c-MET, a positive control) are added. After 48 hours, the culture medium is removed and a rapid detection reagent containing β-gal substrate is added to each well. Hydrolysis of the substrate by β-gal results in the generation of a chemiluminescent signal. After incubation for 1 hour, luminescence is detected on a standard luminescence plate reader (Perkin Elmer).

[0363] Consistent with the literature, currently used clinically cMET antibodies (such as onatumumab) induce robust cMET dimerization at low concentrations. In contrast, despite cMET dimerization, bispecific antibodies require higher concentrations and exhibit lower activation. This data suggests that bispecific antibodies can reduce signaling caused by dimerization compared to currently available monospecific antibodies.

[0364] Additional embodiments

[0365] Embodiment 1: A method for degrading cMET protein on a target cell, the method comprising: contacting cMET protein and a membrane-associated internalization protein on the target cell with a bispecific binding agent, wherein contact of the cMET protein and the membrane-associated internalization protein with the bispecific binding agent results in internalization and degradation of the cMET protein; and wherein the bispecific binding agent comprises: (a) a first binding domain that specifically binds to an extracellular epitope of the membrane-associated internalization protein; and (b) a second binding domain that specifically binds to an extracellular epitope of the cMET protein; wherein the membrane-associated internalization protein is selected from the group consisting of CEACAM5, CEACAM6, HER3, MUC1, CD205, CD166, PRLR, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, CD40, CD228A, MUC5A, CD44, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6 , ADAM9, I-Ag7, ENPP3, CD37, CD46, CD56, CD74, IGF1R, ROR1, CDH6, ROR2, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56 and CD71.

[0366] Embodiment 2: The method of embodiment 1, wherein the membrane-associated internalization protein is selected from the group consisting of CD205, CD166, SLC34A2, ITGB6, LRRC15, MUC16, SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56 and CD71.

[0367] Embodiment 3: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is CEACAM5.

[0368] Embodiment 4: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is CEACAM6.

[0369] Embodiment 5: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is HER3.

[0370] Embodiment 6: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is MUCl.

[0371] Embodiment 7: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is CD205.

[0372] Embodiment 8: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is CD166.

[0373] Embodiment 9: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is PRLR.

[0374] Embodiment 10: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is SLC34A2.

[0375] Embodiment 11: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is ITGB6.

[0376] Embodiment 12: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is LRRC15.

[0377] Embodiment 13: The method of embodiment 1 or embodiment 2, wherein the membrane-associated internalization protein is MUC16.

[0378] Embodiment 14: The method of any one of Embodiments 1 to 13, wherein the bispecific binding agent comprises an antibody or portion thereof.

[0379] Embodiment 15: The method of any one of Embodiments 1 to 13, wherein the bispecific binding agent comprises a bispecific antibody or portion thereof.

[0380] Embodiment 16: The method of any one of Embodiments 1 to 13, wherein the bispecific binding agent comprises a knob-in-hole bispecific IgG.

[0381] Embodiment 17: The method of any one of Embodiments 1 to 13, wherein the bispecific binding agent does not comprise an antibody drug conjugate.

[0382] Embodiment 18: A bispecific binding agent comprising a bispecific antibody or antibody derivative, the bispecific binding agent comprising: a) a first binding domain that specifically binds to an extracellular epitope of a cMET protein on a target cell; and b) a second binding domain that specifically binds to an extracellular epitope of a membrane-associated internalization protein on a target cell; wherein the membrane-associated internalization protein is selected from the group consisting of CD205, CD166, SLC34A2, ITGB6, LRRC15 and MUC16 SLC39A6, AXL, CD40, CD228, MUC5A, ITGB1, STn, KAAG1, DLK1, 5T4, SEZ6, ADAM9, I-Ag7, ENPP3, CD46, CD56, ROR1, GPR20, TM4SF1, B7-H4, ALPP, LY6E, CLDN18, LY6G6D, GPR56 and CD71.

[0383] Embodiment 19: The bispecific binding agent of embodiment 18, wherein the membrane-associated internalization protein is CD205.

[0384] Embodiment 20: The bispecific binding agent of embodiment 18, wherein the membrane-associated internalization protein is CD166.

[0385] Embodiment 21: The bispecific binding agent of Embodiment 18, wherein the membrane-associated internalization protein is SLC34A2.

[0386] Embodiment 22: The bispecific binding agent of embodiment 18, wherein the membrane-associated internalization protein is ITGB6.

[0387] Embodiment 23: The bispecific binding agent of Embodiment 18, wherein the membrane-associated internalization protein is LRRC15.

[0388] Embodiment 24: The bispecific binding agent of Embodiment 18, wherein the membrane-associated internalization protein is MUC16.

[0389] Embodiment 25: The bispecific binding agent of any one of Embodiments 18 to 24, wherein the bispecific binding agent comprises a knob-in-hole bispecific IgG.

[0390] Embodiment 26: The bispecific binding agent of any one of Embodiments 18 to 25, wherein the bispecific binding agent does not comprise an antibody drug conjugate.

[0391] Embodiment 27: A pharmaceutical composition comprising the bispecific binding agent of any one of Embodiments 18 to 26 and a pharmaceutically acceptable excipient.

[0392] Embodiment 28: A method of treating cancer in a subject in need thereof, the method comprising administering to the subject the bispecific binding agent of any one of Embodiments 18 to 26 or the pharmaceutical composition of Embodiment 27.

[0393] Embodiment 29: A method of preventing the growth of a target cell, the method comprising contacting the cell with the bispecific binding agent of any one of embodiments 18 to 26 or the pharmaceutical composition of embodiment 27.

[0394] Embodiment 30: The method of Embodiment 29, wherein the cell is a cancer cell.

Claims

1. A method for degrading a target protein on the surface of a target cell, the method comprising: contacting the degradation protein and the target protein on the surface of the target cell with a binding agent, wherein the binding agent comprises: (i) a first binding domain, which specifically binds to the degradation protein; (ii) a second binding domain that specifically binds to the target protein, wherein the target protein comprises cMET.

2. The method of claim 1, wherein the binding agent is a multispecific antibody, a bispecific antibody, a bispecific diabody, a bispecific Fab2, a bispecific camel antibody, a bispecific peptide antibody scFv-Fc, a bispecific IgG, a knob-in-hole bispecific IgG, an Fc-Fab, or a knob-in-hole bispecific Fc-Fab.

3. The method of claim 2, wherein the binding agent is a multispecific antibody or a bispecific antibody.

4. The method of claim 3, wherein the binding agent is a bispecific antibody.

5. The method of any one of claims 1 to 4, wherein the degraded protein is CDH3, MUC1, CD276, TROP2, CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, EGFR, MST1R, EphA2, ADAM9, IGF1R, RNF43, RNF128, RNF130, or ZNRF3. The method of claim 5 , wherein the degraded protein is CDH3.

7. The method of claim 6, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the variable heavy chain of the first binding domain comprises at least 80% sequence identity to any one of SEQ ID NO: 106, 110, 114, 118, or 122.

8. The method of claim 7, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 106, 110, 114, 118, or 122.

9. The method of claim 8, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 106, 110, 114, 118, or 122.

10. The method of any one of claims 6 to 9, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 108, 112, 116, 120, or 124.

11. The method of claim 10, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 108, 112, 116, 120, or 124.

12. The method of claim 11, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 108, 112, 116, 120, or 124.

13. The method of any one of claims 6 to 12, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

14. The method of claim 13, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

15. The method of any one of claims 6 to 12, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

16. The method of claim 5, wherein the degraded protein is MUCl.

17. The method of claim 16, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 6, 10, 14, 18, or 22.

18. The method of claim 17, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 6, 10, 14, 18, or 22.

19. The method of claim 18, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 6, 10, 14, 18, or 22.

20. The method of any one of claims 16 to 19, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 8, 12, 16, 20, or 24.

21. The method of claim 20, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 8, 12, 16, 20, or 24.

22. The method of claim 21, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 8, 12, 16, 20, or 24.

23. The method of any one of claims 16 to 22, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NO: 6, 10, 14, 18, or 22 and any one of SEQ ID NO: 8, 12, 16, 20, or 24.

24. The method of claim 23, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24.

25. The method of any one of claims 16 to 22, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24.

26. The method of claim 5, wherein the degraded protein is CD276.

27. The method of claim 26, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 25, 26, 30, 34, or 38.

28. The method of claim 27, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 26, 30, 34, or 38.

29. The method of claim 28, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 26, 30, 34, or 38.

30. The method of any one of claims 26 to 29, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 28, 32, 36, or 40.

31. The method of claim 30, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 28, 32, 36, or 40.

32. The method of claim 31, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 28, 32, 36, or 40.

33. The method of any one of claims 26 to 32, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

34. The method of claim 33, wherein the first binding domain binds to an epitope e of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

35. The method of any one of claims 26 to 32, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

36. The method of claim 5, wherein the degradation protein is TROP2.

37. The method of claim 36, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 198, 202, 206, 210, or 214.

38. The method of claim 37, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 198, 202, 206, 210, or 214.

39. The method of claim 38, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 198, 202, 206, 210, or 214.

40. The method of any one of claims 36 to 39, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 200, 204, 208, 212, or 216.

41. The method of claim 40, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 200, 204, 208, 212, or 216.

42. The method of claim 41, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 200, 204, 208, 212, or 216.

43. The method of any one of claims 36 to 42, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

44. The method of claim 43, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

45. The method of any one of claims 36 to 42, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

46. ​​The method of claim 5, wherein the degraded protein is selected from CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, MST1R, EphA2, ADAM9, IGF1R, and EGFR.

47. The method of claim 5, wherein the degraded protein is RNF43, RNF128, RNF130, or ZNRF3.

48. The method of claim 46 or 47, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any of the variable heavy chain sequences listed in Table 1 or Table 4.

49. The method of claim 48, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of the variable heavy chain sequences listed in Table 1 or Table 4.

50. The method of claim 49, wherein the first binding domain variable heavy chain comprises any one of the variable heavy chain sequences listed in Table 1 or Table 4.

51. The method of any one of claims 46 to 50, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of the variable light chain sequences listed in Table 1 or Table 4.

52. The method of claim 51, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of the variable light chain sequences listed in Table 1 or Table 4.

53. The method of claim 52, wherein the first binding domain variable light chain comprises any one of the variable light chain sequences listed in Table 1 or Table 4.

54. The method of any one of claims 46 to 53, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4.

55. The method of claim 54, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4.

56. The method of any one of claims 46 to 53, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of the variable heavy chain sequences or any of the variable light chain sequences listed in Table 1 or Table 4.

57. The method of any one of claims 1 to 56, wherein the second binding domain comprises a second binding domain variable heavy chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364.

58. The method of claim 57, wherein the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364.

59. The method of claim 58, wherein the second binding domain variable heavy chain comprises any one of SEQ ID NO: 352, 356, 360, or 364.

60. The method of any one of claims 1 to 59, wherein the second binding domain comprises a second binding domain variable light chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366.

61. The method of claim 60, wherein the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366.

62. The method of claim 61, wherein the second binding domain variable heavy chain comprises any one of SEQ ID NO: 354, 358, 362, or 366.

63. The method of any one of claims 1 to 62, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by eviantazumab.

64. The method of claim 63, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by eviantazumab.

65. The method of any one of claims 1 to 62, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope to which eviantazumab binds.

66. The method of any one of claims 1 to 65, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by terituzumab.

67. The method of claim 66, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by terituzumab.

68. The method of any one of claims 1 to 65, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope to which terituzumab binds.

69. The method of any one of claims 1 to 68, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to the epitope bound by onatumumab.

70. The method of claim 69, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by onatumumab.

71. The method of any one of claims 1 to 68, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by onatumumab.

72. The method of any one of claims 1 to 71, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to the epitope bound by REGN5093s58.

73. The method of claim 72, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by REGN5093s58.

74. The method of any one of claims 1 to 71, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by REGN5093s58.

75. The method of any one of claims 1 to 74, wherein after the contacting, cMET is internalized into the target cell along with the degradation protein, and cMET is degraded.

76. The method of claim 75, wherein after the cMET is internalized into the target cell together with the degradation protein, the degradation protein is recycled to the surface of the target cell.

77. The method of claim 75, wherein after the cMET is internalized into the target cell along with the degradation protein, one or more of the degradation protein or the cMET is degraded.

78. The method of any one of claims 1 to 77, wherein the target cell is a cancer cell.

79. The method of claim 78, wherein the cancer cells are selected from breast cancer cells, B cell lymphoma cells, pancreatic cancer cells, Hodgkin lymphoma cells, ovarian cancer cells, prostate cancer cells, mesothelioma cells, lung cancer cells, non-Hodgkin B cell (B-NHL) cells, melanoma cells, chronic lymphocytic leukemia cells, acute lymphocytic leukemia cells, neuroblastoma cells, glioma cells, glioblastoma cells, bladder cancer cells, colorectal cancer cells, gastric adenocarcinoma cells, non-small cell lung cancer cells, and head and neck cancer cells, and cancers carrying cMET mutations including exon 14 deletions.

80. The method of claim 79, wherein the cancer cells are gastric adenocarcinoma cells.

81. The method of claim 79, wherein the cancer cells are non-small cell lung cancer cells.

82. The method of claim 79, wherein the cancer cell comprises a genetic mutation selected from a cMET exon 14 skipping mutation or a cMET duplication mutation.

83. The method of claim 82, wherein the mutation comprises a cMET exon 14 skipping mutation.

84. The method of claim 82, wherein the cancer cell comprises a cMET repeat mutation.

85. The method of any one of claims 78 to 84, wherein expression of cMET in the cancer cells after the contacting with the binding agent is less than expression of cMET in a control cancer cell not contacted with the binding agent.

86. The method of claim 85, wherein expression of cMET in the cancer cells after the contacting with the binding agent is at least 50% less than expression of cMET in a control cancer cell not contacted with the binding agent.

87. The method of any one of claims 78 to 84, wherein expression of cMET in the cancer cells following the contacting with the binding agent is at least 50% less than expression of cMET in a control cancer cell contacted with the monospecific cMET binding agent.

88. The method of any one of claims 78 to 87, wherein the amount of cMET on the surface of the cancer cell after the contacting with the binding agent is less than the amount of cMET on the surface of a control cancer cell not contacted with the binding agent.

89. The method of claim 88, wherein the amount of cMET on the surface of the cancer cell after the contacting with the binding agent is at least 20% less than the amount of cMET on the surface of a control cancer cell not contacted with the binding agent.

90. The method of any one of claims 78 to 87, wherein the amount of cMET on the surface of the cancer cell after said contacting with the binding agent is at least 20% less than the amount of cMET on the surface of a control cancer cell contacted with a monospecific cMET binding agent.

91. The method of any one of claims 88 to 90, wherein the amount of cMET on the surface of the cancer cells is determined by staining the cancer cells with a fluorescently labeled antibody against cMET and measuring fluorescence intensity.

92. The method of any one of claims 78 to 91, wherein the amount of cMET internalized by the cancer cell after the contacting with the binding agent is greater than the amount of cMET internalized by a control cancer cell not contacted with the binding agent.

93. The method of claim 92, wherein the amount of cMET internalized by the cancer cell after the contacting with the binding agent is at least 20% greater than the amount of cMET internalized by a control cancer cell not contacted with the binding agent.

94. The method of any one of claims 78 to 91, wherein the amount of cMET internalized by the cancer cell following the contacting with the binding agent is at least 20% greater than the amount of cMET internalized by a control cancer cell contacted with the monospecific cMET binding agent.

95. The method of any one of claims 92 to 94, wherein the amount of internalized cMET is determined by labeling the binding agent with a fluorescent tag prior to contacting the degraded protein and the target protein and measuring the fluorescence of the fluorescent tag after contacting the degraded protein and the target protein, wherein the fluorescent tag selectively fluoresces at intracellular pH.

96. The method of any one of claims 78 to 95, wherein the amount of cMET degraded in the cancer cell after the contacting with the binding agent is greater than the amount of cMET degraded in a control cancer cell not contacted with the binding agent.

97. The method of claim 96, wherein the amount of cMET degraded in the cancer cell after the contacting with the binding agent is at least 20% greater than the amount of cMET degraded in a control cancer cell not contacted with the binding agent.

98. The method of any one of claims 78 to 95, wherein the amount of cMET degraded in the cancer cell after the contacting with the binding agent is at least 20% greater than the amount of cMET degraded in a control cancer cell contacted with the monospecific cMET binding agent.

99. The method of any one of claims 78 to 98, wherein the amount of cMET dimer on the cancer cell after the contact with the binding agent is less than the amount of cMET dimer on a control cancer cell not contacted with the binding agent.

100. The method of any one of claims 78 to 98, wherein the amount of cMET dimer on the cancer cell after the contacting with the binding agent is less than the amount of cMET dimer on a control cancer cell contacted with a monospecific cMET binding agent.

101. The method of any one of claims 78 to 100, wherein the amount of cMET activation in the cancer cell following the contacting with the binding agent is within 50% of the amount of cMET activation in a control cancer cell not contacted with the binding agent.

102. The method of any one of claims 90, 94, 98, or 100, wherein the monospecific cMET-binding agent is terituzumab.

103. The method of any one of claims 90, 94, 98, or 100, wherein the monospecific cMET-binding agent is onatumumab.

104. The method of any one of claims 90, 94, 98, or 100, wherein the monospecific cMET-binding agent is REGN5093s58.

105. The method of any one of claims 78 to 104, wherein the method increases the sensitivity of the cancer cells to a cancer therapeutic agent or radiation therapy.

106. The method of claim 105, wherein the cancer therapeutic agent is a cytotoxic agent.

107. The method of any one of claims 78 to 106, wherein the method reduces proliferation of the cancer cells.

108. The method of any one of claims 78 to 107, wherein the method induces death of the cancer cell.

109. The method of any one of claims 1 to 108, wherein the contacting is performed in vivo.

110. A method of treating cancer in a subject in need thereof, the method comprising: administering to the subject a binding agent, wherein the binding agent comprises: (i) a first binding domain that specifically binds to a degradation protein, wherein the degradation protein is expressed on a target cell; (ii) a second binding domain that specifically binds to the target protein, wherein the target protein comprises cMET.

111. The method of claim 110, wherein the binding agent is a multispecific antibody, a bispecific antibody, a bispecific diabody, a bispecific Fab2, a bispecific camelid antibody, a bispecific peptide antibody scFv-Fc, a bispecific IgG, a knob-in-hole bispecific IgG, an Fc-Fab, or a knob-in-hole bispecific Fc-Fab.

112. The method of claim 111, wherein the binding agent is a multispecific antibody or a bispecific antibody.

113. The method of claim 112, wherein the binding agent is a bispecific antibody.

114. The method of any one of claims 110 to 113, wherein the degraded protein is CDH3, MUC1, CD276, TROP2, CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, EGFR, MST1R, EphA2, ADAM9, IGF1R, RNF43, RNF128, RNF130, or ZNRF3.

115. The method of claim 114, wherein the degraded protein is CDH3.

116. The method of claim 115, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the variable heavy chain of the first binding domain comprises at least 80% sequence identity to any one of SEQ ID NO: 106, 110, 114, 118, or 122.

117. The method of claim 116, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 106, 110, 114, 118, or 122.

118. The method of claim 117, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 106, 110, 114, 118, or 122.

119. The method of any one of claims 115 to 118, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 108, 112, 116, 120, or 124.

120. The method of claim 119, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

121. The method of claim 120, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 108, 112, 116, 120, or 124.

122. The method of any one of claims 115 to 121, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

123. The method of claim 122, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NO: 106, 110, 114, 118, or 122 and any one of SEQ ID NO: 108, 112, 116, 120, or 124.

124. The method of any one of claims 115 to 121, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

125. The method of claim 114, wherein the degraded protein is MUCl.

126. The method of claim 125, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 6, 10, 14, 18, or 22.

127. The method of claim 126, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 6, 10, 14, 18, or 22.

128. The method of claim 127, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 6, 10, 14, 18, or 22.

129. The method of any one of claims 125 to 128, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 8, 12, 16, 20, or 24.

130. The method of claim 129, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 8, 12, 16, 20, or 24.

131. The method of claim 130, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 8, 12, 16, 20, or 24.

132. The method of any one of claims 125 to 131, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NO: 6, 10, 14, 18, or 22 and any one of SEQ ID NO: 8, 12, 16, 20, or 24.

133. The method of claim 132, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NO: 6, 10, 14, 18, or 22 and any one of SEQ ID NO: 8, 12, 16, 20, or 24.

134. The method of any one of claims 125 to 131, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 6, 10, 14, 18, or 22 and any one of SEQ ID NOs: 8, 12, 16, 20, or 24.

135. The method of claim 114, wherein the degraded protein is CD276.

136. The method of claim 135, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 26, 30, 34, or 38.

137. The method of claim 136, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 26, 30, 34, or 38.

138. The method of claim 137, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 26, 30, 34, or 38.

139. The method of any one of claims 135 to 138, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 28, 32, 36, or 40.

140. The method of claim 139, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 28, 32, 36, or 40.

141. The method of claim 140, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 28, 32, 36, or 40.

142. The method of any one of claims 135 to 141, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

143. The method of claim 142, wherein the first binding domain binds to an epitope e of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

144. The method of any one of claims 135 to 141, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

145. The method of claim 114, wherein the degradation protein is TROP2.

146. The method of claim 145, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 198, 202, 206, 210, or 214.

147. The method of claim 146, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 198, 202, 206, 210, or 214.

148. The method of claim 147, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 198, 202, 206, 210, or 214.

149. The method of any one of claims 145 to 148, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 200, 204, 208, 212, or 216.

150. The method of claim 149, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 200, 204, 208, 212, or 216.

151. The method of claim 150, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 200, 204, 208, 212, or 216.

152. The method of any one of claims 145 to 151, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

153. The method of claim 152, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

154. The method of any one of claims 145 to 151, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

155. The method of claim 114, wherein the degraded protein is selected from CD71, HER3, TNFRSF10B, ITGB6, PD-L1, EpCAM, TPBG, MST1R, EphA2, ADAM9, IGF1R, and EGFR.

156. The method of claim 114, wherein the degraded protein is RNF43, RNF128, RNF130, or ZNRF3.

157. The method of claim 155 or 156, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity with any of the variable heavy chain sequences listed in Table 1 or Table 4.

158. The method of claim 157, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any of the variable heavy chain sequences listed in Table 1 or Table 4.

159. The method of claim 158, wherein the first binding domain variable heavy chain comprises any one of the variable heavy chain sequences listed in Table 1 or Table 4.

160. The method of any one of claims 155 to 159, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any of the variable light chain sequences listed in Table 1 or Table 4.

161. The method of claim 160, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any of the variable light chain sequences listed in Table 1 or Table 4.

162. The method of claim 161, wherein the first binding domain variable light chain comprises any one of the variable light chain sequences listed in Table 1 or Table 4.

163. The method of any one of claims 155 to 162, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4.

164. The method of claim 163, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with an epitope bound by an antibody comprising any one of the variable heavy chain sequences or any one of the variable light chain sequences listed in Table 1 or Table 4.

165. The method of any one of claims 155 to 162, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any of the variable heavy chain sequences or any of the variable light chain sequences listed in Table 1 or Table 4.

166. The method of any one of claims 110 to 165, wherein the second binding domain comprises a second binding domain variable heavy chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364.

167. The method of claim 166, wherein the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364.

168. The method of claim 167, wherein the second binding domain variable heavy chain comprises any one of SEQ ID NO: 352, 356, 360, or 364.

169. The method of any one of claims 110 to 168, wherein the second binding domain comprises a second binding domain variable light chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366.

170. The method of claim 169, wherein the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366.

171. The method of claim 170, wherein the second binding domain variable heavy chain comprises any one of SEQ ID NO: 354, 358, 362, or 366.

172. The method of any one of claims 110 to 171, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by evitumomab.

173. The method of claim 172, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by eviantazumab.

174. The method of any one of claims 110 to 171, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope to which eviantazumab binds.

175. The method of any one of claims 110 to 174, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by terituzumab.

176. The method of claim 175, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by terituzumab.

177. The method of any one of claims 110 to 174, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope to which terituzumab binds.

178. The method of any one of claims 110 to 177, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by onatumumab.

179. The method of claim 178, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by onatumumab.

180. The method of any one of claims 110 to 177, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by onatumumab.

181. The method of any one of claims 110 to 180, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by REGN5093s58.

182. The method of claim 181, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by REGN5093s58.

183. The method of any one of claims 110 to 180, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by REGN5093s58.

184. The method of any one of claims 110 to 183, wherein the cancer cell is selected from the group consisting of breast cancer cells, B-cell lymphoma cells, pancreatic cancer cells, Hodgkin lymphoma cells, ovarian cancer cells, prostate cancer cells, mesothelioma cells, lung cancer cells, non-Hodgkin B cells (B-NHL) cells, melanoma cells, chronic lymphocytic leukemia cells, acute lymphocytic leukemia cells, neuroblastoma cells, glioma cells, glioblastoma cells, bladder cancer cells, colorectal cancer cells, gastric adenocarcinoma cells, non-small cell lung cancer cells, and head and neck cancer cells.

185. The method of claim 184, wherein the cancer cells are gastric adenocarcinoma cells.

186. The method of claim 184, wherein the cancer cells are non-small cell lung cancer cells.

187. The method of claim 184, wherein the cancer cell comprises a genetic mutation selected from a cMET exon 14 skipping mutation or a cMET duplication mutation.

188. The method of claim 187, wherein the mutation comprises a cMET exon 14 skipping mutation.

189. The method of claim 187, wherein the cancer cells comprise a cMET repeat mutation.

190. The method of any one of claims 110 to 189, wherein the method increases the sensitivity of cancer cells to a cancer therapeutic agent or radiation therapy.

191. The method of claim 184, wherein the cancer therapeutic agent is a cytotoxic agent.

192. The method of any one of claims 110 to 191, wherein the method reduces proliferation of cancer cells.

193. The method of any one of claims 110 to 192, wherein the method induces death of cancer cells.

194. A binding agent comprising: A first binding domain that specifically binds to a degradation protein, wherein the degradation protein is CDH3, MUC1, CD276, or TROP2; and A second binding domain that specifically binds to a target protein, wherein the target protein is cMET.

195. The binding agent of claim 194, wherein the multispecific binding agent is a multispecific antibody, a bispecific antibody, a bispecific diabody, a bispecific Fab2, a bispecific camelid antibody, a bispecific peptibody scFv-Fc, a bispecific IgG, a knob-in-hole bispecific IgG, an Fc-Fab, or a knob-in-hole bispecific Fc-Fab.

196. The binding agent of claim 195, wherein the binding agent is a multispecific antibody or a bispecific antibody.

197. The binding agent of claim 196, wherein the binding agent is a bispecific antibody.

198. The binding agent of any one of claims 194 to 197, wherein the degradation protein is CDH3.

199. The binding agent of claim 198, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the variable heavy chain of the first binding domain comprises at least 80% sequence identity with any one of SEQ ID NO: 106, 110, 114, 118 or 122.

200. The binding agent of claim 199, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 106, 110, 114, 118 or 122.

201. The binding agent of claim 200, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 106, 110, 114, 118 or 122.

202. The binding agent of any one of claims 198 to 201, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 108, 112, 116, 120, or 124.

203. The binding agent of claim 202, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 108, 112, 116, 120 or 124.

204. The binding agent of claim 203, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 108, 112, 116, 120, or 124.

205. The binding agent of any one of claims 198 to 204, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NO: 106, 110, 114, 118, or 122 and any one of SEQ ID NO: 108, 112, 116, 120, or 124.

206. The binding agent of claim 205, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NO: 106, 110, 114, 118 or 122 and any one of SEQ ID NO: 108, 112, 116, 120 or 124.

207. The binding agent of any one of claims 198 to 204, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 106, 110, 114, 118, or 122 and any one of SEQ ID NOs: 108, 112, 116, 120, or 124.

208. The binding agent of any one of claims 194 to 197, wherein the degradation protein is MUCl.

209. The binding agent of claim 208, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 6, 10, 14, 18 or 22.

210. The binding agent of claim 209, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 6, 10, 14, 18 or 22.

211. The binding agent of claim 210, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 6, 10, 14, 18 or 22.

212. The binding agent of any one of claims 207 to 211, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 8, 12, 16, 20, or 24.

213. The binding agent of claim 212, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 8, 12, 16, 20 or 24.

214. The binding agent of claim 213, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 8, 12, 16, 20, or 24.

215. The binding agent of any one of claims 207 to 214, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NO: 6, 10, 14, 18, or 22 and any one of SEQ ID NO: 8, 12, 16, 20, or 24.

216. A binding agent as described in claim 215, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by the following antibody: the antibody comprises any one of SEQ ID NO: 6, 10, 14, 18 or 22 and any one of SEQ ID NO: 8, 12, 16, 20 or 24.

217. The binding agent of any one of claims 207 to 214, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NO: 6, 10, 14, 18, or 22 and any one of SEQ ID NO: 8, 12, 16, 20, or 24.

218. The binding agent of any one of claims 194 to 197, wherein the degraded protein is CD276.

219. The binding agent of claim 218, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 26, 30, 34 or 38.

220. The binding agent of claim 219, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 26, 30, 34 or 38.

221. The binding agent of claim 220, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 26, 30, 34 or 38.

222. The binding agent of any one of claims 218 to 221, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 28, 32, 36, or 40.

223. The binding agent of claim 222, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 28, 32, 36 or 40.

224. The binding agent of claim 223, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 28, 32, 36 or 40.

225. The binding agent of any one of claims 218 to 224, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

226. A binding agent as described in claim 225, wherein the first binding domain binds to an epitope e of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with the epitope bound by the following antibody: the antibody comprises any one of SEQ ID NO: 26, 30, 34 or 38 and any one of SEQ ID NO: 28, 32, 36 or 40.

227. The binding agent of any one of claims 218 to 224, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 26, 30, 34, or 38 and any one of SEQ ID NOs: 28, 32, 36, or 40.

228. The binding agent of any one of claims 194 to 197, wherein the degradation protein is TROP2.

229. The binding agent of claim 228, wherein the first binding domain comprises a first binding domain variable heavy chain, and wherein the first binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NO: 198, 202, 206, 210 or 214.

230. The binding agent of claim 229, wherein the first binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 198, 202, 206, 210 or 214.

231. The binding agent of claim 230, wherein the first binding domain variable heavy chain comprises any one of SEQ ID NO: 198, 202, 206, 210, or 214.

232. The binding agent of any one of claims 228 to 231, wherein the first binding domain comprises a first binding domain variable light chain, and wherein the first binding domain variable light chain comprises at least 80% sequence identity to any one of SEQ ID NO: 200, 204, 208, 212, or 216.

233. The binding agent of claim 232, wherein the first binding domain variable light chain comprises at least 90% sequence identity to any one of SEQ ID NO: 200, 204, 208, 212 or 216.

234. The binding agent of claim 233, wherein the first binding domain variable light chain comprises any one of SEQ ID NO: 200, 204, 208, 212 or 216.

235. The binding agent of any one of claims 228 to 234, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

236. The binding agent of claim 235, wherein the first binding domain binds to an epitope of the degraded protein on the target cell, wherein the epitope comprises at least 90% sequence identity with an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

237. The binding agent of any one of claims 228 to 234, wherein the first binding domain binds to an epitope of the degraded protein on the target cell that does not comprise any of the amino acids from an epitope bound by an antibody comprising any one of SEQ ID NOs: 198, 202, 206, 210, or 214 and any one of SEQ ID NOs: 200, 204, 208, 212, or 216.

238. The binding agent of any one of claims 194 to 237, wherein the second binding domain comprises a second binding domain variable heavy chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 352, 356, 360, or 364.

239. The binding agent of claim 238, wherein the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 352, 356, 360 or 364.

240. The binding agent of claim 239, wherein the second binding domain variable heavy chain comprises any one of SEQ ID NO: 352, 356, 360 or 364.

241. The binding agent of any one of claims 194 to 240, wherein the second binding domain comprises a second binding domain variable light chain, and wherein the second binding domain variable heavy chain comprises at least 80% sequence identity to any one of SEQ ID NOs: 354, 358, 362, or 366.

242. The binding agent of claim 241, wherein the second binding domain variable heavy chain comprises at least 90% sequence identity to any one of SEQ ID NO: 354, 358, 362 or 366.

243. The binding agent of claim 242, wherein the second binding domain variable heavy chain comprises any one of SEQ ID NO: 354, 358, 362 or 366.

244. The binding agent of any one of claims 194 to 243, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by evitumomab.

245. The binding agent of claim 244, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by eviantazumab.

246. The binding agent of any one of claims 194 to 243, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope to which eviantazumab binds.

247. The binding agent of any one of claims 194 to 246, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to the epitope bound by terituzumab.

248. The binding agent of claim 247, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to an epitope bound by terituzumab.

249. The binding agent of any one of claims 194 to 246, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope to which terituzumab binds.

250. The binding agent of any one of claims 194 to 249, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to an epitope bound by onatumumab.

251. The binding agent of claim 250, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by onatumumab.

252. The binding agent of any one of claims 194 to 249, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by onatumumab.

253. The binding agent of any one of claims 194 to 252, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 80% sequence identity to the epitope bound by REGN5093s58.

254. The binding agent of claim 253, wherein the second binding domain binds to an epitope of the target protein on the target cell, wherein the epitope comprises at least 90% sequence identity to the epitope bound by REGN5093s58.

255. The binding agent of any one of claims 194 to 252, wherein the second binding domain binds to an epitope of the target protein on the target cell that does not comprise any of the amino acids from the epitope bound by REGN5093s58.

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