Tumor activating antibodies targeting TROP2 and uses thereof

Abstract

Disclosed herein are isolated polypeptides or polypeptide complexes comprising a tumor-associated calcium signal transduction protein 2 (TROP2) binding domain having optimized binding and kinetic properties. In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 338,172, filed May 4, 2022, the entire contents of which are incorporated herein by reference.

[0003] Sequence Listing

[0004] This application contains a sequence listing submitted electronically in XML format, the entire contents of which are hereby incorporated by reference. The XML copy was created on April 27, 2023, named 52426-742_601SL.xml, and is 487,415 bytes in size. Summary of the Invention

[0005] Disclosed herein are isolated polypeptides and polypeptide complexes according to Formula I: A1-L1-P1 (Formula I), wherein A1 comprises a recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2. NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YWX 14 X 15 X 16 X 17 wherein X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17 is V, G, P, L, I, M, S, T, or A; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linker portion that connects A1 to P1 and is a substrate for a tumor-specific protease. In some embodiments, X1 is Q, N, D, E, or A; X2 is Q, N, D, E, or A; X3 is I, V, L, or A; X4 is T, S, or A; X5 is P, G, or A; X6 is L, V, I, or A; X7 is T, S, or A; X8 is R, K, or A; X9 is G, S, T, or A; X 10 Is F, Y or A; X 11 is G, S, T or A; X 12 is S, G, T or A; X 13 is S, G, T or A; X 14 is Y, W, F or A; X 15 is F, Y, W or A; X 16 is D, E, Q, N, or A; and X 17 is V, L, I or A. In some embodiments, X1 is Q; and X6 is L. In some embodiments, X8 is R; X 10 It's F; X 11 It is G; X 14 It is Y; X 15 is F; and X 16 It’s D.

[0006] In some embodiments, X1 is Q, S, T, D, N, E, or A; X2 is Q, S, T, D, N, E, or A; X3 is I, G, P, V, L, M, or A; X4 is T, G, S, M, H, N, Q, or A; X5 is P, G, V, L, I, M, or A; X6 is L, G, P, V, I, M, or A; X7 is T, G, S, M, H, N, Q, or A; X8 is R, H, K, or A; X9 is G, P, V, L, I, M, S, T, or A; X 10is F, Y, W, V, L, I, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q or A; X 13 is S, G, T, M, N, Q or A; X 14 is Y, F, W, V, L, I, or A; X 15 is F, Y, W, V, L, I, or A; X 16 is D, Q, N, E, S, T, or A; and X 17 is V, G, P, L, I, M, or A. In some embodiments, X1 is Q, N, or A; X2 is Q, N, or A; X3 is I, V, L, or A; X4 is T, S, or A; X5 is P, G, or A; X6 is L, V, I, or A; X7 is T, S, or A; X8 is R, K, or A; X9 is G, V, S, T, or A; X 10 Is F, Y or A; X 11 is G, V, S, T or A; X 12 is S, G, T or A; X 13 is S, G, T or A; X 14 is Y, W or A; X 15 Is F, Y or A; X 16 is D, E, or A; and X 17 is V, G, L, I or A. In some embodiments, X1 is Q; and X6 is L. In some embodiments, X8 is R; X 10 It's F; X 11 It is G; X 14 It is Y; X 15 is F; and X 16 It’s D.

[0007] In some embodiments, CDR3-L comprises an amino acid selected from SEQ ID NOs: 3-5 and 8-12. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 5, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 12 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15.In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 5, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 9, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15.

[0008] In some embodiments, CDR3-H comprises amino acids selected from SEQ ID NOs: 16-17, 19-22, and 25-28. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14; CDR3-H: SEQ ID NO: 16; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 17; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 25;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:27; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:28. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 17; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 21; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:22; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:28. ;

[0009] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 16, CDR2-L: SEQ ID NO: 17, CDR3-H: SEQ ID NO: 18; NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2;CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2; CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14; CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163;and L1 comprises a linker portion that connects A1 to P1 and is a substrate for a tumor-specific protease. In some embodiments, the TROP2 binding domain comprises a Fab, Fab', (Fab')2, or a single-chain variable fragment (scFv). In some embodiments, the TROP2 binding domain is a Fab. In some embodiments, the immunoglobulin light chain comprises the variable domain of an immunoglobulin kappa (IgK) or immunoglobulin lambda (IgL) light chain. In some embodiments, the immunoglobulin heavy chain comprises the variable domain of an IgG1, IgG2, IgG3, or IgG4 heavy chain. ;

[0010] In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NO: 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, or 73. In some embodiments, the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NO: 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, or 74. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 33, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 34. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 35, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 36. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 37, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:40.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 41, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 42. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 43, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 44. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 47, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 48. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 50. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 51, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:53, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:54.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 55, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 56. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 57, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 58. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 62. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 63, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 64. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 65, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:67, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:68.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:73, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:74.

[0011] In some embodiments, the TROP2 binding domain has weaker binding to TROP2 as measured by ELISA under substantially similar assay conditions than a TROP2 binding domain comprising an immunoglobulin light chain according to SEQ ID NO: 29 and an immunoglobulin heavy chain according to SEQ ID NO: 30. In some embodiments, the TROP2 binding domain has an increased EC for TROP2 as measured by ELISA under substantially similar assay conditions than a TROP2 binding domain comprising an immunoglobulin light chain according to SEQ ID NO: 29 and an immunoglobulin heavy chain according to SEQ ID NO: 30. 50 In some embodiments, the TROP2 binding domain has a faster off-rate (greater k) for TROP2 binding than a TROP2 binding domain comprising an immunoglobulin light chain according to SEQ ID NO: 29 and an immunoglobulin heavy chain according to SEQ ID NO: 30, as measured under substantially similar kinetic assay conditions. diss ).

[0012] In some embodiments, P1 weakens the binding of A1 to TROP2. In some embodiments, P1 binds to A1 through ionic interactions, electrostatic interactions, hydrophobic interactions, Pi stacking interactions, and H-bond interactions, or a combination thereof. In some embodiments, P1 binds to A1 at or near the antigen binding site. In some embodiments, when L1 is cleaved by a tumor-specific protease, P1 dissociates from A1, thereby exposing A1 to TROP2. In some embodiments, P1 has less than 75% sequence identity with TROP2. In some embodiments, P1 has less than 80% sequence identity with TROP2. In some embodiments, P1 has less than 85% sequence identity with TROP2. In some embodiments, P1 has less than 90% sequence identity with TROP2. In some embodiments, P1 has less than 95% sequence identity with TROP2. In some embodiments, P1 comprises a de novo generated amino acid sequence that shares less than 10% sequence identity with TROP2. In some embodiments, P1 comprises at least two cysteine ​​amino acid residues. In some embodiments, P1 comprises a cyclic peptide or a linear peptide. In some embodiments, P1 comprises a cyclic peptide. In some embodiments, P1 comprises a linear peptide. In some embodiments, P1 comprises a modified amino acid or an unnatural amino acid, or a modified unnatural amino acid, or a combination thereof. In some embodiments, P1 does not comprise albumin or an albumin fragment. In some embodiments, P1 does not comprise an albumin binding domain.

[0013] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 102, 107, 123, and 124, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 102, 107, 123, and 124. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, wherein P1 comprises an amino acid sequence according to SEQ ID NO: 102 or SEQ ID NO: 107, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 102 or SEQ ID NO: 107. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 29, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 30, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 102, 107, 123, and 124, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 102, 107, 123, and 124. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:29, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:30, and wherein P1 comprises an amino acid sequence according to SEQ ID NO:102 or SEQ ID NO;107, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to SEQ ID NO:102 or SEQ ID NO:107.

[0014] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 40, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 40, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150.

[0015] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 16, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 16, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:50, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 50, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 142, and 150.

[0016] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 22, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 22, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 141, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 62, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:62, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:107, 141, 142 and 150.

[0017] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 26, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 26, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 26, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 141, 142, and 150.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 70, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 70, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 141, 142, and 150.

[0018] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 27, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 27, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, 158, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, 158. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 72, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:72, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158.

[0019] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain. In some embodiments, the isolated polypeptide or polypeptide complex is according to the following formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75; CDR2-L comprises the amino acid sequence of SEQ ID NO: 76; CDR3-L comprises the amino acid sequence of SEQ ID NO: 77; CDR1-H comprises the amino acid sequence of SEQ ID NO: 78; CDR2-H comprises the amino acid sequence of SEQ ID NO: 79; and CDR3-H comprises the amino acid sequence of SEQ ID NO: 80.

[0020] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H; wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75 or SEQ ID NO: 259; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: NO:76 amino acid sequence; wherein CDR3-L comprises the amino acid sequence of Z1-Z2-W-Z3-Z4-Z5-Z6-W-Z7-Z8; wherein Z1 is V, G, P, L, I, M, S, T or A; Z2 is L, G, P, V, I, M, S, T or A; Z3 is Y, F, W, V, L, I, G or A; Z4 is S, G, T, M, N, Q, H or A; Z5 is N, Q, S, T, D, E, H, K, R or A; Z6 is R, S, T, Q, D, E, H, K, N or A; Z7 is V, G, P, L, I, M, S, T or A; and Z8 is F, Y, W, V, L, I, G or A; wherein CDR1-H comprises SEQ ID NO:78 or SEQ ID NO: NO:270 amino acid sequence; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:79; and wherein CDR3-H comprises Z9-Z 10 -Z 11 -Z 12 -NZ 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -YZ 20 -AZ 21 wherein Z9 is V, G, P, L, I, M, S, T or A; Z 10 is R, S, T, Q, D, E, H, K, N or A; Z 11 is H, R, K, G, T, S, N, Q or A; Z 12 is G, P, V, L, I, M, S, T, or A; Z 13 is F, Y, W, V, L, I, G, or A; Z 14 is G, P, V, L, I, M, S, T, or A; Z 15 is N, Q, S, T, D, E, H, K, R, or A; Z 16 is S, G, T, M, N, Q, H or A; Z 17 Y, F, W, V, L, I, G, or A; Z 18 is I, G, P, V, L, M, S, T, or A; Z19 is S, G, T, M, N, Q, H or A; Z 20 is W, F, Y, V, L, I, G, or A; and Z 21 is Y, F, W, V, L, I, G, or A. In some embodiments, Z1 is V, G, L, I, or A; Z2 is L, V, I, or A; Z3 is Y, W, F, or A; Z4 is S, G, T, or A; Z5 is N, Q, D, E, or A; Z6 is R, K, or A; Z7 is V, G, L, I, or A; Z8 is F, Y, W, or A; Z9 is V, G, L, I, or A; Z 10 R, K or A; Z 12 is G, S, T or A; Z 13 Is F, Y, W or A; Z 14 is G, S, T or A; Z 15 is N, Q, D, E or A; Z 16 Is S, G, T or A; Z 17 Y, W, F or A; Z 18 Is I, V, L or A; Z 19 Is S, G, T or A; Z 20 is W, Y, F, or A; and Z 21 is Y, W, F or A. In some embodiments, Z8 is F. In some embodiments, Z 10 It is R; Z 11 It is H; Z 13 It's F; Z 18 It is I; Z 19 is S; and Z 20 It’s W.

[0021] In some embodiments, the CDR3-L of the CD3 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 77, 260-261, 263-266, and 268-269. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 263, and CDR1-H: SEQ ID NO: NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQID NO:76, CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:268 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80;and CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:269 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 263, and CDR1-H: SEQ ID NO: NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQID NO:76, CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80;and CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 268 and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80. ;

[0022] In some embodiments, CDR3-H of the CD3 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 80, 271-274, 276-282, and 284-285. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 271; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 272; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 272. NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:273; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:274; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:276; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:281; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:282; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:284; and CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:285.;

[0023] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 271; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 274; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 274. NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; and CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:285.

[0024] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 81. ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:262 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:263 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:267 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:268 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:269 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:271; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77, CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:272; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:273; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:274; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:275;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:276; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:281; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:282; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:283;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:284; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:285; and CDR1-L: SEQ ID NO:259, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:260 and CDR1-H: SEQ ID NO:270, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:286. ;

[0025] In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises the variable domain of an immunoglobulin kappa (IgK) or immunoglobulin lambda (IgL) light chain. In some embodiments, the immunoglobulin heavy chain of the CD3 binding domain comprises the variable domain of an IgG1, IgG2, IgG3, or IgG4 heavy chain. In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 81. In some embodiments, the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 82. In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:81, and the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:82.

[0026] In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 99. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 303. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 304. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 306. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 307. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 309. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 310. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 311. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 312. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 313. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:314.In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 315. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 316. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 317. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 318. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 319. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 320. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 321. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 322. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 323. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 324. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 325. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 326. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:327.In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 328. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329.

[0027] In some embodiments, the CD3 binding domain comprises Fab, Fab', (Fab')2 or a single-chain variable fragment (scFv). In some embodiments, the CD3 binding domain is a scFv. In some embodiments, the N-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the C-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the N-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the N-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the C-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the N-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the N-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the N-terminus of the immunoglobulin light chain of the TROP2 binding domain binds to the C-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin light chain of the TROP2 binding domain is bound to the N-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the N-terminus of the immunoglobulin light chain of the TROP2 binding domain is bound to the C-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin light chain of the TROP2 binding domain is bound to the N-terminus of the immunoglobulin heavy chain of the CD3 binding domain.

[0028] In some embodiments, the CD3 binding domain is a scFv and the TROP2 binding domain is a Fab or Fab'. In some embodiments, the scFv binds to the immunoglobulin heavy chain of Fab or Fab'. In some embodiments, the scFv binds to the immunoglobulin light chain of Fab or Fab'. In some embodiments, the immunoglobulin light chain of the scFv binds to the immunoglobulin heavy chain of Fab or Fab'. In some embodiments, the immunoglobulin light chain of the scFv binds to the immunoglobulin light chain of Fab or Fab'. In some embodiments, the immunoglobulin heavy chain of the scFv binds to the immunoglobulin heavy chain of Fab or Fab'. In some embodiments, the immunoglobulin heavy chain of the scFv binds to the immunoglobulin light chain of Fab or Fab'.

[0029] In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and SEQ ID NO: 88. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 89 and SEQ ID NO: 90. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 91 and SEQ ID NO: 92. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 93 and SEQ ID NO: 94. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95 and SEQ ID NO: 96. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 97 and SEQ ID NO: 98.

[0030] In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 330 and SEQ ID NO: 331. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 332 and SEQ ID NO: 333. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 332 and SEQ ID NO: 333. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 334 and SEQ ID NO: 335. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 336 and SEQ ID NO: 337. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 338 and SEQ ID NO: 339. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 340 and SEQ ID NO: 341. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 342 and SEQ ID NO: 343. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 344 and SEQ ID NO: 345. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:346 and SEQ ID NO:347.In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 348 and SEQ ID NO: 349. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 350 and SEQ ID NO: 351.

[0031] In some embodiments, the recombinant antibody or antigen-binding fragment thereof has weaker cytotoxic activity than a recombinant antibody or antigen-binding fragment thereof comprising an immunoglobulin light chain according to SEQ ID NO: 83 or 85 and an immunoglobulin heavy chain according to SEQ ID NO: 84 or 86, as measured in an in vitro tumor cell killing assay under substantially similar assay conditions.

[0032] In some embodiments, P2 weakens the binding of B2 to CD3. In some embodiments, P2 binds to B2 through ionic interactions, electrostatic interactions, hydrophobic interactions, Pi stacking interactions, and H-bond interactions, or a combination thereof. In some embodiments, P2 binds to B2 at or near the antigen binding site. In some embodiments, when L2 is cleaved by a tumor-specific protease, P2 dissociates from B2, thereby exposing B2 to CD3. In some embodiments, P2 has less than 70% sequence identity with CD3. In some embodiments, P2 has less than 75% sequence identity with CD3. In some embodiments, P2 has less than 80% sequence identity with CD3. In some embodiments, P2 has less than 85% sequence identity with CD3. In some embodiments, P2 has less than 90% sequence identity with CD3. In some embodiments, P2 has less than 95% sequence identity with CD3. In some embodiments, P2 comprises a de novo generated amino acid sequence that shares less than 10% sequence identity with CD3. In some embodiments, P2 comprises a peptide sequence of at least 5 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 6 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 10 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 10 amino acids in length and no more than 20 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 16 amino acids in length. In some embodiments, P2 comprises a peptide sequence of no more than 40 amino acids in length. In some embodiments, P2 comprises at least two cysteine ​​amino acid residues. In some embodiments, P2 comprises a cyclic peptide or a linear peptide. In some embodiments, P2 comprises a cyclic peptide. In some embodiments, P2 comprises a linear peptide. In some embodiments, P2 comprises a modified amino acid or a non-natural amino acid, or a modified non-natural amino acid, or a combination thereof. In some embodiments, P2 does not comprise albumin or an albumin fragment. In some embodiments, P2 does not comprise an albumin binding domain. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO: 292.

[0033] In some embodiments, P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 287-302, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 287-302. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 273, and wherein P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 273, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 273, and wherein P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305, and wherein P2 comprises the amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298.In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence according to SEQ ID NO: 305, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295.

[0034] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 276, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 276, and wherein P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308, and wherein P2 comprises the amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence according to SEQ ID NO: 308, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 259, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, CDR1-H: SEQ ID NO: 270, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 286, and wherein P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298.In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 259, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, CDR1-H: SEQ ID NO: 270, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 286, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 259, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, CDR1-H: SEQ ID NO: 270, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 286, and wherein P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329, and wherein P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298. In some embodiments, the CD3 binding domain comprises an amino acid sequence according to SEQ ID NO: 329, and wherein P2 comprises an amino acid sequence according to SEQ ID NO: 295.

[0035] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:261, CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, and CDR3-H: SEQ ID NO:80, and wherein P2 comprises an amino acid sequence according to SEQ ID NO:295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO:295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 80, and wherein P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 319, and wherein P2 comprises the amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises the amino acid sequence according to SEQ ID NO:319, and wherein P2 comprises the amino acid sequence according to SEQ ID NO:295.

[0036] In some embodiments, L1 or L2 is a peptide sequence having at least 5 to no more than 50 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 10 to no more than 30 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 10 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 18 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 26 amino acids. In some embodiments, L1 or L2 comprises (G2S) n wherein n is an integer from 1 to 3. In some embodiments, L1 or L2 comprises (G2S) n wherein n is an integer of at least 1. In some embodiments, L1 or L2 comprises a compound selected from (G2S) n , (GS) n、(GSGGS) n 、(GGGS) n 、(GGGGS) n and (GSSGGS) n wherein n is an integer of at least 1. In some embodiments, the tumor-specific protease is selected from a metalloprotease, a serine protease, a cysteine ​​protease, a threonine protease, and an aspartic protease. In some embodiments, L1 or L2 comprises an amino acid sequence cleavable by urokinase, an amino acid sequence cleavable by a membrane-type serine protease (matriptase), or an amino acid sequence cleavable by a matrix metalloprotease. In some embodiments, L1 or L2 comprises a sequence according to any one of SEQ ID NOs: 226-254. In some embodiments, L1 is bound to the N-terminus of A1. In some embodiments, L1 is bound to the C-terminus of A1. In some embodiments, L2 is bound to the N-terminus of B2. In some embodiments, L2 is bound to the C-terminus of B2.

[0037] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a half-life extending molecule (H1). In some embodiments, H1 is linked to P1. In some embodiments, H1 is linked to P2. In some embodiments, H1 does not block the CD3 binding domain from CD3. In some embodiments, the half-life extending molecule (H1) does not have binding affinity for CD3. In some embodiments, the half-life extending molecule (H1) does not shield the isolated polypeptide or polypeptide complex from CD3. In some embodiments, H1 comprises a sequence according to SEQ ID NOs: 255-258. In some embodiments, H1 comprises an amino acid sequence having a repeating sequence motif. In some embodiments, H1 comprises an amino acid sequence having a highly ordered secondary structure. In some embodiments, H1 comprises a polymer. In some embodiments, the polymer is polyethylene glycol (PEG). In some embodiments, H1 comprises albumin. In some embodiments, H1 comprises an Fc domain. In some embodiments, the albumin is serum albumin. In some embodiments, the albumin is human serum albumin. In some embodiments, H1 comprises a polypeptide, a ligand, or a small molecule. In some embodiments, the polypeptide, ligand, or small molecule binds to a serum protein or fragment thereof, a circulating immunoglobulin or fragment thereof, or CD35 / CR1. In some embodiments, the serum protein comprises thyroxine-binding protein, transthyretin, 1-acid glycoprotein, transferrin, transferrin receptor or a transferrin-binding portion thereof, fibrinogen, or albumin. In some embodiments, the circulating immunoglobulin molecule comprises IgG1, IgG2, IgG3, IgG4, slgA, IgM, or IgD. In some embodiments, the serum protein is albumin. In some embodiments, the polypeptide is an antibody. In some embodiments, the antibody comprises a single domain antibody, a single chain variable fragment, or a Fab. In some embodiments, the single domain antibody comprises a single domain antibody that binds to albumin. In some embodiments, the single domain antibody is a human antibody or a humanized antibody. In some embodiments, the single domain antibody is selected from 645gH1gL1, 645dsgH5gL4, 23-13-A01-sc02, A10m3 or a fragment thereof, DOM7r-31, DOM7h-11-15, Alb-1, Alb-8, Alb-23, 10G, 10E, and SA21.In some embodiments, the single-domain antibody comprises a complementarity determining region (CDR): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, HC-CDR2, and HC-CDR3 of the single-domain antibody comprise: HC-CDR1: SEQ ID NO: 255, HC-CDR2: SEQ ID NO: 256, and HC-CDR3: SEQ ID NO: 257; and wherein the CDRs comprise 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, H1 comprises an amino acid sequence according to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises a modified amino acid or a non-natural amino acid, or a modified non-natural amino acid, or a combination thereof. In some embodiments, the modified amino acid or the modified non-natural amino acid comprises a post-translational modification. In some embodiments, H1 comprises a linking portion (L3) connecting H1 to P1 or P2. In some embodiments, L3 is a peptide sequence having at least 5 to no more than 50 amino acids. In some embodiments, L3 is a peptide sequence having at least 10 to no more than 30 amino acids. In some embodiments, L3 is a peptide sequence having at least 10 amino acids. In some embodiments, L3 is a peptide sequence having at least 18 amino acids. In some embodiments, L3 is a peptide sequence having at least 26 amino acids. In some embodiments, L3 comprises a peptide selected from (G2S). n , (GS) n 、(GSGGS) n 、(GGGS) n 、(GGGGS) n and (GSSGGS) n wherein n is an integer of at least 1.

[0038] In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 166 and SEQ ID NO: 167. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 168 and SEQ ID NO: 169. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 170 and SEQ ID NO: 171. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 172 and SEQ ID NO: 173. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 174 and SEQ ID NO: 175. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 176 and SEQ ID NO: 177. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 178 and SEQ ID NO: 179. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 180 and SEQ ID NO: 181. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 182 and SEQ ID NO: 183. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 184 and SEQ ID NO: 185.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 186 and SEQ ID NO: 187. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 188 and SEQ ID NO: 189. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 190 and SEQ ID NO: 191. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 192 and SEQ ID NO: 193. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 194 and SEQ ID NO: 195. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 196 and SEQ ID NO: 197. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 198 and SEQ ID NO: 199. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 200 and SEQ ID NO: 201. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 202 and SEQ ID NO: 203. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:204 and SEQ ID NO:205.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 206 and SEQ ID NO: 207. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 208 and SEQ ID NO: 209. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 210 and SEQ ID NO: 211. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 212 and SEQ ID NO: 213. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 214 and SEQ ID NO: 215. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 216 and SEQ ID NO: 217. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 218 and SEQ ID NO: 219. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 220 and SEQ ID NO: 221. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 222 and SEQ ID NO: 223. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:224 and SEQ ID NO:225.

[0039] In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 352 and SEQ ID NO: 353. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 354 and SEQ ID NO: 355. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 356 and SEQ ID NO: 357. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 358 and SEQ ID NO: 359. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 360 and SEQ ID NO: 361. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 362 and SEQ ID NO: 363. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 364 and SEQ ID NO: 365. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 366 and SEQ ID NO: 367. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 368 and SEQ ID NO: 369. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:370 and SEQ ID NO:371.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 372 and SEQ ID NO: 373. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 374 and SEQ ID NO: 375. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 376 and SEQ ID NO: 377. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 378 and SEQ ID NO: 379. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 380 and SEQ ID NO: 381. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 382 and SEQ ID NO: 383. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 384 and SEQ ID NO: 385. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 386 and SEQ ID NO: 387. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 388 and SEQ ID NO: 389. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:390 and SEQ ID NO:391.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 392 and SEQ ID NO: 393. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 394 and SEQ ID NO: 395.

[0040] Disclosed herein are isolated polypeptides and polypeptide complexes according to Formula I: A1-L1-P1 (Formula I), wherein A1 comprises a recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; wherein CDR3-L comprises the amino acid sequence of SEQ ID NO: 3; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and wherein CDR3-H comprises the amino acid sequence of SEQ ID NO: 15. NO:26 amino acid sequence; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119, and 123-163; and L1 comprises a linker that connects A1 to P1 and is a substrate for a tumor-specific protease. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs:107, 141, 142, and 150. In some embodiments, P1 comprises an amino acid sequence according to SEQ ID NO: 142. In some embodiments, the TROP2 binding domain comprises Fab, Fab', (Fab')2 or a single-chain variable fragment (scFv). In some embodiments, the TROP2 binding domain is Fab. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 85% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 85% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 70.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 92% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 92% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises the amino acid sequence of SEQ ID NO: 69, and the immunoglobulin heavy chain comprises the amino acid sequence of SEQ ID NO: 70.

[0041] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain. In some embodiments, the isolated polypeptide or polypeptide complex is according to the formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 80.

[0042] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 276. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 259, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 260, CDR1-H comprises the amino acid sequence of SEQ ID NO: 270, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 286. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 273.

[0043] In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 81, and the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 82. In some embodiments, the CD3 binding domain comprises a Fab, Fab', (Fab')2, or a single-chain variable fragment (scFv). In some embodiments, the CD3 binding domain is an scFv. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 99. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305.

[0044] In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO: 292. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 295. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 200 and SEQ ID NO: 201. In some embodiments, the isolated polypeptide or polypeptide complex comprises the amino acid sequence of SEQ ID NO: 200 and SEQ ID NO: 201. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 362 and SEQ ID NO: 363. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 382 and SEQ ID NO: 383. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 390 and SEQ ID NO: 391.

[0045] Disclosed herein are isolated polypeptides and polypeptide complexes according to Formula I: A1-L1-P1 (Formula I), wherein A1 comprises a recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; wherein CDR3-L comprises the amino acid sequence of SEQ ID NO: 3; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and wherein CDR3-H comprises the amino acid sequence of SEQ ID NO: 15. NO:27; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163; and L1 comprises a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158. In some embodiments, P1 comprises an amino acid sequence according to SEQ ID NO: 107.

[0046] In some embodiments, the TROP2 binding domain comprises Fab, Fab', (Fab')2 or a single-chain variable fragment (scFv). In some embodiments, the TROP2 binding domain is Fab. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 92% identity to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 92% identity to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises the amino acid sequence of SEQ ID NO: 71, and the immunoglobulin heavy chain comprises the amino acid sequence of SEQ ID NO: 72.

[0047] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain. In some embodiments, the isolated polypeptide or polypeptide complex is according to the formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 80.

[0048] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 276. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 259, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 260, CDR1-H comprises the amino acid sequence of SEQ ID NO: 270, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 286.

[0049] In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 81, and the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 82. In some embodiments, the CD3 binding domain comprises a Fab, Fab', (Fab')2, or a single-chain variable fragment (scFv). In some embodiments, the CD3 binding domain is an scFv. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 99. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329.

[0050] In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO: 292. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 295. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 180 and SEQ ID NO: 181. In some embodiments, the isolated polypeptide or polypeptide complex comprises the amino acid sequence of SEQ ID NO: 180 and SEQ ID NO: 181. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 352 and SEQ ID NO: 353. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:354 and SEQ ID NO:355.

[0051] Disclosed herein are pharmaceutical compositions comprising (i) the isolated polypeptide or polypeptide complex thereof according to any one of the above embodiments, and (ii) a pharmaceutically acceptable excipient.

[0052] Disclosed herein are isolated recombinant nucleic acid molecules encoding the isolated polypeptide or polypeptide complex of any of the above embodiments.

[0053] Disclosed herein are methods of treating cancer in a subject in need thereof, comprising administering to the subject an isolated polypeptide or polypeptide complex of any one of the embodiments described above. In some embodiments, the cancer comprises breast cancer, lung cancer, urothelial cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, gastric cancer, colon cancer, head and neck cancer, or glioma. In some embodiments, the breast cancer comprises triple-negative breast cancer. In some embodiments, the lung cancer comprises non-small cell lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The novel features of the present disclosure are particularly set forth 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 of illustrative embodiments utilizing the principles of the present disclosure and the accompanying drawings, in which:

[0055] Figure 1A-1J Shown are biolayer interferometry (BLI) titration data of TROP2 binding of non-mutated and mutated TROP2 binding domains (TBDs).

[0056] Figure 2A-2M Shown are BLI titration data of TROP2 binding of mutant TBDs.

[0057] Figure 3 Figure 2 shows the TROP2 binding curves and binding EC for non-mutated TBD and mutant TBD as measured by ELISA. 50 .

[0058] Figure 4 Figure 2 shows the TROP2 binding curves and binding EC for non-mutated TBD and mutant TBD as measured by ELISA. 50 .

[0059] Figure 5A-5G Shown are binding curves of peptide binding to TBD-1 as measured by ELISA.

[0060] Figures 6A-6F Shown are binding curves of peptide binding to TBD-6 as measured by ELISA.

[0061] Figures 7A-7F Shown are binding curves of peptide binding to TBD-11 as measured by ELISA.

[0062] Figures 8A-8D Shown are binding curves of peptide binding to TBD-17 as measured by ELISA.

[0063] Figures 9A-9D Shown are binding curves of peptide binding to TBD-21 as measured by ELISA.

[0064] Figures 10A-10D Shown are binding curves of peptide binding to TBD-22 as measured by ELISA.

[0065] Figures 11A-11G Graph showing inhibition of TBD-1 binding to TROP2 by the peptides of the present disclosure.

[0066] Figures 12A-12F Graph showing inhibition of TBD-6 binding to TROP2 by the peptides of the present disclosure.

[0067] Figures 13A-13F Graph showing inhibition of TBD-11 binding to TROP2 by the peptides of the present disclosure.

[0068] Figures 14A-14F Graph showing inhibition of TBD-17 binding to TROP2 by peptides of the present disclosure.

[0069] Figures 15A-15F Graph showing inhibition of TBD-21 binding to TROP2 by peptides of the present disclosure.

[0070] Figures 16A-16F Graph showing inhibition of TBD-22 binding to TROP2 by peptides of the present disclosure.

[0071] Figures 17A-17D Shown are BLI titration data of TROP2 binding of non-mutated and mutated T cell engagers (TCEs).

[0072] Figures 18A-18D Shown are BLI titration data of TROP2 binding of non-mutated TCE and mutated TCE.

[0073] Figures 19A-19B Shown is the TROP2 ( Figure 19A ) and CD3ε( Figure 19B ) Binding curves and binding EC 50 .

[0074] Figure 20A-Figure 20B Shown is the TROP2 ( Figure 20A ) and CD3e( Figure 20B ) Binding curves and binding EC 50 .

[0075] Figures 21A-21B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 21A) and CD3ε( Figure 21B ) Binding curves and binding EC 50 .

[0076] Figures 22A-22B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 22A ) and CD3ε( Figure 22B ) Binding curves and binding EC 50 .

[0077] Figures 23A-23B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 23A ) and CD3ε( Figure 23B ) Binding curves and binding EC 50 .

[0078] Figures 24A-24B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 24A ) and CD3ε( Figure 24B ) Binding curves and binding EC 50 .

[0079] Figures 25A-25B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 25A ) and CD3ε( Figure 25B ) Binding curves and binding EC 50 .

[0080] Figures 26A-26B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 26A ) and CD3ε( Figure 26B ) Binding curves and binding EC 50 .

[0081] Figures 27A-27B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 27A ) and CD3ε( Figure 27B ) Binding curves and binding EC 50 .

[0082] Figures 28A-28B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 28A ) and CD3ε( Figure 28B ) Binding curves and binding EC 50 .

[0083] Figure 29A-29B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 29A ) and CD3ε( Figure 29B ) Binding curves and binding EC 50 .

[0084] Figure 30A-Figure 30B Shown is the TROP2 of TCE and polypeptide complex as measured by ELISA ( Figure 30A ) and CD3ε( Figure 30B ) Binding curves and binding EC 50 .

[0085] Figure 31 Shown are the inhibition of aCD3 scFv binding to CD3 by peptides as measured by ELISA.

[0086] Figures 32A-32J The figure shows the killing of TROP2-positive HCT116 tumor cells by the polypeptide complex in the presence of CD8+ T cells.

[0087] Figures 33A-33P The graph shows the killing of TROP2-positive MDAMB231 tumor cells by the peptide complex in the presence of CD8+ T cells.

[0088] Figures 34A-34T The graph shows the killing of TROP2-positive NCI-H292 tumor cells by the peptide complex in the presence of CD8+ T cells.

[0089] Figure 35 The graph shows the killing of recombinant HEK293 cells expressing human TROP2 by the polypeptide complex in the presence of CD8+ T cells.

[0090] Figure 36 The graph shows the killing of recombinant HEK293 cells expressing cynomolgus monkey TROP2 by the polypeptide complex in the presence of CD8+ T cells.

[0091] Figure 37 Figure 2 shows tumor cell viability of wild-type HEK293 cells in the presence of CD8+ T cells with increasing concentrations of peptide complexes.

[0092] Figures 38A-38I Shown are the peptide pharmacokinetics in cynomolgus monkeys following a single IV bolus injection.

[0093] Figures 39A-39F Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-5.

[0094] Figure 40 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-1.

[0095] Figure 41 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-2.

[0096] Figure 42 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-3.

[0097] Figure 43 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-9.

[0098] Figure 44 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-15.

[0099] Figure 45 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-6.

[0100] Figure 46 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-8.

[0101] Figure 47 Shown is cytokine release in cynomolgus monkeys following a single IV bolus of the polypeptide complex PC-18.

[0102] Figure 48 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys following a single IV bolus of PC-5.

[0103] Figure 49 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys following a single IV bolus of PC-1.

[0104] Figure 50 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys following a single IV bolus of PC-2.

[0105] Figure 51 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys following a single IV bolus of PC-3.

[0106] Figure 52 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys following a single IV bolus of PC-9.

[0107] Figure 53 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys after a single intravenous injection of PC-15.

[0108] Figure 54 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys after a single intravenous injection of PC-6.

[0109] Figure 55 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys after a single intravenous injection of PC-8.

[0110] Figure 56 Shown are serum liver enzymes, bilirubin, and urea in cynomolgus monkeys after a single intravenous injection of PC-18.

[0111] Figure 57 Comparison of in vitro cytotoxicity changes and clinical chemistry safety signals in cynomolgus monkeys for PC-9, PC-15, PC-6, PC-5, PC8, and PC-18 is shown.

[0112] Figure 58 Shown are polypeptide pharmacokinetics in cynomolgus monkeys following continuous IV infusion of TCE-1.

[0113] Figure 59 Shown are the polypeptide pharmacokinetics in cynomolgus monkeys following continuous IV infusion of TCE-8 at different dosing rates.

[0114] Figure 60 Shown are the polypeptide pharmacokinetics in cynomolgus monkeys following continuous IV infusion of TCE-7 at different dosing rates.

[0115] Figure 61 Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-1.

[0116] Figure 62 Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-8 at different dosing rates.

[0117] Figure 63 Shown are cytokine release in cynomolgus monkeys following continuous infusion of TCE-7 at different dosing rates.

[0118] Figure 64-Figure 65 Shown are the results of non-human primate (NHP) toxicity studies of PC-8 and TCE-8.

[0119] Figure 66-Figure 67 Shown are the results of NHP toxicity studies of PC-18 and TCE-7 in cynomolgus monkeys.

[0120] Figure 68 Figure 2 shows the human CD3 binding curves and binding EC of TCE-1 and TCE-9 to TCE-18 as measured by ELISA. 50 .

[0121] Figure 69 Figure 2 shows the human CD3 binding curves and binding EC of TCE-1 and TCE-19 to TCE-28 as measured by ELISA. 50 .

[0122] Figure 70 Figure 2 shows the human CD3 binding curves and binding EC of TCE-1 and TCE-29 to TCE-35 as measured by ELISA. 50 .

[0123] Figure 71 Shown are binding curves of peptide binding to TCE-1 as measured by ELISA.

[0124] Figure 72 Shown are binding curves of peptide binding to TCE-10 as measured by ELISA.

[0125] Figure 73 Shown are binding curves of peptide binding to TCE-11 as measured by ELISA.

[0126] Figure 74 Shown are binding curves of peptide binding to TCE-13 as measured by ELISA.

[0127] Figure 75 Shown are binding curves of peptide binding to TCE-14 as measured by ELISA.

[0128] Figure 76 Shown are binding curves of peptide binding to TCE-20 as measured by ELISA.

[0129] Figure 77 Shown are binding curves of peptide binding to TCE-21 as measured by ELISA.

[0130] Figure 78 Shown are binding curves of peptide binding to TCE-22 as measured by ELISA.

[0131] Figure 79 Shown are binding curves of peptide binding to TCE-26 as measured by ELISA.

[0132] Figure 80 Shown are binding curves of peptide binding to TCE-31 as measured by ELISA.

[0133] Figure 81 Shown are binding curves of peptide binding to TCE-33 as measured by ELISA.

[0134] Figure 82 Shown are binding curves of peptide binding to TCE-34 as measured by ELISA.

[0135] Figure 83 Shown are binding curves of peptide binding to TCE-35 as measured by ELISA.

[0136] Figures 84A-84C Shown are the binding curves of peptide-73 binding to TROP2 TCE as measured by ELISA.

[0137] Figure 85 Shown are binding curves of peptide binding to TCE-7 as measured by ELISA.

[0138] Figure 86 Shown are binding curves of peptide binding to TCE-39 as measured by ELISA.

[0139] Figure 87 Shown are binding curves of peptide binding to TCE-40 as measured by ELISA.

[0140] Figure 88 Shown are binding curves of peptide binding to TCE-36 as measured by ELISA.

[0141] Figure 89 Shown are binding curves of peptide binding to TCE-37 as measured by ELISA.

[0142] Figure 90 Shown are binding curves of peptide binding to TCE-38 as measured by ELISA.

[0143] Figure 91 Graph depicts inhibition of TCE-1 binding to CD3 by peptides as measured by ELISA.

[0144] Figure 92 Graph depicts inhibition of TCE-10 binding to CD3 by peptides as measured by ELISA.

[0145] Figure 93 Graph depicts inhibition of TCE-11 binding to CD3 by peptides as measured by ELISA.

[0146] Figure 94 Graph depicts inhibition of TCE-13 binding to CD3 by peptides as measured by ELISA.

[0147] Figure 95 Graph depicts inhibition of TCE-14 binding to CD3 by peptides as measured by ELISA.

[0148] Figure 96 Graph depicts inhibition of TCE-20 binding to CD3 by peptides as measured by ELISA.

[0149] Figure 97 Graph depicts inhibition of TCE-22 binding to CD3 by peptides as measured by ELISA.

[0150] Figure 98 Graph depicts inhibition of TCE-26 binding to CD3 by peptides as measured by ELISA.

[0151] Figure 99 Graph depicts inhibition of TCE-31 binding to CD3 by peptides as measured by ELISA.

[0152] Figure 100 Graph depicts inhibition of TCE-33 binding to CD3 by peptides as measured by ELISA.

[0153] Figure 101 Graphed is the inhibition of TCE-34 binding to CD3 by peptides as measured by ELISA.

[0154] Figure 102 Graph depicts inhibition of TCE-35 binding to CD3 by peptides as measured by ELISA.

[0155] Figures 103A-103C Graph depicts the inhibition of TCE binding to CD3 by peptide-73 as measured by ELISA.

[0156] Figure 104 Graph depicts inhibition of TCE-7 binding to CD3 by peptides as measured by ELISA.

[0157] Figure 105 Graph depicts inhibition of TCE-39 binding to CD3 by peptides as measured by ELISA.

[0158] Figure 106 Graph depicts inhibition of TCE-40 binding to CD3 by peptides as measured by ELISA.

[0159] Figure 107 Graph depicts inhibition of TCE-36 binding to CD3 by peptides as measured by ELISA.

[0160] Figure 108 Graph depicts inhibition of TCE-37 binding to CD3 by peptides as measured by ELISA.

[0161] Figure 109 Graph depicts inhibition of TCE-38 binding to CD3 by peptides as measured by ELISA.

[0162] Figure 110 Shown are binding curves for TROP2 binding of TCE as measured by ELISA.

[0163] Figure 111 Shown are the binding curves for CD3 binding of TCE as measured by ELISA.

[0164] Figure 112 Shown are binding curves for TROP2 binding of TCE as measured by ELISA.

[0165] Figure 113 Shown are the binding curves for CD3 binding of TCE as measured by ELISA.

[0166] Figure 114 Shown are binding curves of TROP2 binding of TCE and PC as measured by ELISA.

[0167] Figure 115 Shown are binding curves for CD3 binding of TCE and PC as measured by ELISA.

[0168] Figure 116 Shown are binding curves of TROP2 binding of TCE and PC as measured by ELISA.

[0169] Figure 117 Shown are binding curves for CD3 binding of TCE and PC as measured by ELISA.

[0170] Figure 118 Shown are binding curves of TROP2 binding of TCE and PC as measured by ELISA.

[0171] Figure 119 Shown are binding curves for CD3 binding of TCE and PC as measured by ELISA.

[0172] Figure 120 Shown are binding curves of TROP2 binding of TCE and PC as measured by ELISA.

[0173] Figure 121 Shown are binding curves for CD3 binding of TCE and PC as measured by ELISA.

[0174] Figure 122 Shown are binding curves of TROP2 binding of TCE and PC as measured by ELISA.

[0175] Figure 123 Shown are binding curves for CD3 binding of TCE and PC as measured by ELISA.

[0176] Figure 124 Shown are binding curves of TROP2 binding of TCE and PC as measured by ELISA.

[0177] Figure 125 Shown are binding curves for CD3 binding of TCE and PC as measured by ELISA.

[0178] Figures 126-129 Graph depicts killing of TROP2-positive HCT116 tumor cells by TCE and PC in the presence of CD8+ T cells.

[0179] Figures 130-146 Graph depicts killing of TROP2-positive H292 tumor cells by TCE and PC in the presence of CD8+ T cells.

[0180] Figures 147-162 Graph depicts killing of TROP2-expressing cynomolgus monkey HEK293 tumor cells by TCE and PC in the presence of CD8+ T cells.

[0181] Figure 163 The graph shows the results of the pharmacokinetic and toxicity studies of TCE-8 in cynomolgus monkeys.

[0182] Figure 164 The graph shows the results of the pharmacokinetic and toxicity studies of TCE-7 in cynomolgus monkeys.

[0183] Figure 165 Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-8.

[0184] Figure 166 Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-7.

[0185] Figures 167A-167F Shown are clinical chemistry parameters in cynomolgus monkeys following continuous IV infusion of TCE-8.

[0186] Figures 168A-168F Shown are clinical chemistry parameters in cynomolgus monkeys following continuous IV infusion of TCE-7.

[0187] Figure 169 Schematic representation of the pharmacokinetic and toxicity studies of TCE-37 in cynomolgus monkeys.

[0188] Figure 170 Schematic representation of the pharmacokinetic and toxicity studies of TCE-40 in cynomolgus monkeys.

[0189] Figure 171 Schematic representation of the pharmacokinetic and toxicity studies of TCE-38 in cynomolgus monkeys.

[0190] Figures 172A-172D Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-37.

[0191] Figures 173A-173D Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-40.

[0192] Figure 174Shown are cytokine release in cynomolgus monkeys following continuous IV infusion of TCE-38.

[0193] Figures 175A-175F Shown are clinical chemistry parameters in cynomolgus monkeys following continuous IV infusion of TCE-37.

[0194] Figures 176A-176F Shown are clinical chemistry parameters in cynomolgus monkeys following continuous IV infusion of TCE-40.

[0195] Figures 177A-177F Shown are clinical chemistry parameters in cynomolgus monkeys following continuous IV infusion of TCE-38. DETAILED DESCRIPTION

[0196] TROP2 (also known as tumor-associated calcium signal transducer 2 or epithelial glycoprotein-1) is the protein product of the TACSTD2 gene and is a transmembrane glycoprotein that plays a role in a variety of cell signaling pathways, many of which are associated with tumorigenesis. TROP2 has been shown to be overexpressed in a variety of human cancers, including lung cancer, breast cancer, cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer and gastric cancer, and its expression is associated with poor patient prognosis. In addition, TROP2 acts as an oncogene that can drive tumorigenesis and metastasis in epithelial cancers such as colorectal cancer. TROP2 expression in cancer cells has long been associated with drug resistance, and high levels of TROP2 expression have been shown to be associated with poor prognosis in a variety of cancer types. In a meta-analysis comprising data from approximately 2,500 patients, increased TROP2 expression was associated with poor overall survival and disease-free survival outcomes in several solid tumors.

[0197] Disclosed herein are recombinant antibodies or antigen-binding fragments thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain that has been optimized for certain properties. In some embodiments, the TROP2 binding domain has TROP2 binding properties that improve safety and reduce accumulation in healthy tissues. For example, in some embodiments, the TROP2 binding domain has a faster dissociation rate (k) for TROP2 binding. diss ) and / or weaker (higher) EC 50 , which may help reduce accumulation in healthy tissues. In addition, in some embodiments, the TROP2 binding domain has a faster dissociation rate (k diss ) and / or weaker (higher) EC 50 , while maintaining TROP2 binding ability and activity. In some embodiments, the recombinant antibody or antigen-binding fragment thereof exhibits weaker cytotoxic activity to help reduce off-tumor on-target toxicity in healthy tissues.

[0198] Disclosed herein are recombinant antibodies or antigen-binding fragments thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain that have been optimized by alanine scanning mutagenesis. The TROP2 binding domain is alanine scanned to optimize certain properties of the recombinant antibody or antigen-binding fragment thereof. Alanine scanning can be accomplished by systematically mutating individual residues to alanine in the TROP2 binding domain relative to the non-mutated or "wild-type" TROP2 binding domain. Specifically, residues in the third complementarity determining region (CDR3) of the immunoglobulin light chain and the immunoglobulin heavy chain are systematically mutated to alanine to establish CDR3-related structure activity relationships (SARs) and identify TROP2 binding domains that exhibit faster dissociation rates to reduce accumulation in healthy tissues while maintaining binding and activity.

[0199] Disclosed herein are recombinant antibodies or antigen-binding fragments thereof comprising a TROP2-binding domain paired with a peptide that impairs binding of the TROP2-binding domain to TROP2 in healthy tissue. The peptide is linked to the TROP2-binding domain via a linker that is a substrate for a tumor-specific protease. In the tumor microenvironment, the linker is cleaved by the tumor-specific protease, thereby releasing the peptide.

[0200] In some embodiments, the TROP2 binding domain is connected to the CD3 binding domain, which is also paired with a peptide mask that binds to the CD3 binding domain and weakens the binding of the CD3 binding domain to CD3 in healthy tissue. The peptide mask is connected to the CD3 binding domain via a connecting portion that is a substrate for a tumor-specific protease. In the tumor microenvironment, the connecting portion is cleaved by the tumor-specific protease, thereby releasing the peptide mask from the CD3 binding domain. The applicant has found that in some embodiments, the mutation in the CD3 binding domain reduces the binding affinity to CD3, but increases the binding affinity to the peptide mask, thereby improving the masking efficiency. The stronger masking efficiency is manifested as a greater difference in the cell killing efficacy between the non-masked T cell adapter (TCE) containing a non-mutated CD3 binding domain and the masked polypeptide complex compared to the cell killing efficacy difference between the non-masked TCE construct containing a specific CD3 binding domain mutation and the masked polypeptide complex.

[0201] In some embodiments, a half-life extending moiety (such as an anti-albumin domain) is linked to a peptide paired with a CD3 binding domain. In some embodiments, the polypeptides or polypeptide complexes described herein are optimized for a strong TROP2 binding domain and peptide pair that weakens the binding of the TROP2 binding domain to TROP2 in healthy tissue while optimizing a faster dissociation rate of the TROP2 binding domain from TROP2.

[0202] In some embodiments, a recombinant antibody or antigen fragment comprising a TROP2 binding domain that has been optimized for certain properties further comprises a CD3 binding domain. Multispecific antibodies for redirecting T cells to cancer have shown promise in both preclinical and clinical studies. The method relies on the binding of one antigen-interacting portion of the antibody to a tumor-associated antigen or marker (such as TROP2), while the second antigen-interacting portion can bind to an effector cell antigen (such as CD3) on the T cell, which then triggers cytotoxic activity. Disclosed herein are multispecific antibody compositions targeting TROP2 and CD3 for triggering cytotoxic activity.

[0203] In some embodiments, the recombinant antibodies or antigen-binding fragments thereof herein are used in methods for treating cancer. In some embodiments, cancer has cells expressing TROP2. In some cases, cancer is a solid tumor cancer. In some embodiments, cancer is lung cancer, breast cancer (eg, HER2+; ER / PR+; TNBC), cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer, or gastric cancer. In some embodiments, the polypeptide or polypeptide complex herein is used to treat triple-negative breast cancer (TNBC), urothelial carcinoma (UC), non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), gastric cancer, esophageal cancer, head and neck cancer, prostate cancer, or endometrial cancer. In some embodiments, the polypeptide or polypeptide complex herein is used to treat breast cancer, lung cancer, urothelial carcinoma, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, gastric cancer, colon cancer, head and neck cancer, and glioma.

[0204] Certain definitions

[0205] The terms used herein are for the purpose of describing a particular situation only and are not intended to be limiting. As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. Furthermore, to the extent the terms "including," "having," "has," "with," or variations thereof are used in the detailed description and / or claims, these terms are intended to be inclusive in a manner similar to the term "comprising."

[0206] The term "antibody" is used in the broadest sense to cover fully assembled antibodies; antibody fragments that can bind antigen, such as Fab, F(ab')2, Fv; single-chain antibodies (scFv); diabodies; antibody chimeras; hybrid antibodies; bispecific antibodies, etc.

[0207] The term "complementarity determining region" or "CDR" is a segment of an antibody variable region that is structurally complementary to the epitope to which the antibody binds and is more variable than the rest of the variable region. For this reason, CDRs are sometimes referred to as hypervariable regions. The variable region comprises three CDRs. CDR peptides can be obtained by constructing genes encoding the CDRs of the antibody of interest. For example, these genes are prepared by synthesizing the variable regions from RNA of antibody-producing cells using the polymerase chain reaction. See, e.g., Larrick et al., Methods: A Companion to Methods in Enzymology 2: 106 (1991); Courtenay-Luck, “Genetic Manipulation of Monoclonal Antibodies,” in Monoclonal Antibodies: Production, Engineering and Clinical Application, Ritter et al. (Eds.), pp. 166-179 (Cambridge University Press 1995); and Ward et al., “Genetic Manipulation and Expression of Antibodies,” in Monoclonal Antibodies: Principles and Applications, Birch et al., (Eds.), pp. 137-185 (Wiley-Liss, Inc. 1995).

[0208] The term "Fab" refers to a protein containing a light chain constant domain and the first constant domain (CH1) of the heavy chain. Fab fragments differ from Fab' fragments by the addition of a few residues to the carboxyl terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH, referred to herein as Fab', wherein the cysteine ​​residues of the constant domains bear a free thiol group. Fab' fragments are generated by reducing the heavy chain disulfide bridges of the F(ab')2 fragment. Other chemical couplings of antibody fragments are also known.

[0209] "Single-chain variable fragment (scFv)" is a fusion protein of the variable regions of the heavy chain (VH) and light chain (VL) of an antibody, connected by a short linker peptide of ten to about 25 amino acids. For flexibility, the linker is usually rich in glycine, while for solubility, the linker is usually rich in serine or threonine, and the N-terminus of VH can be connected to the C-terminus of VL, and vice versa. This protein retains the specificity of the original antibody, despite the removal of the constant region and the introduction of a linker. scFv antibodies are described, for example, in Houston, JS, Methods in Enzymol. 203 (1991) 46-96). In addition, the antibody fragment comprises a single-chain polypeptide having the characteristics of a VH domain, i.e., capable of being assembled together with a VL domain, or having the characteristics of a VL domain, i.e., capable of being assembled together with a VH domain to form a functional antigen binding site, thereby providing the antigen binding properties of a full-length antibody.

[0210] As used herein, term " percentage (%) amino acid sequence identity " about sequence is defined as, after comparing sequences and introducing room (if necessary) to realize maximum percentage sequence identity and any conservative substitution is not considered as a part for sequence identity, the percentage of the amino acid residue identical with the amino acid residue in the particular sequence in the candidate sequence.Comparison for determining percentage amino acid sequence identity can be realized in a variety of ways well known by persons skilled in the art, for example, using publicly available computer software such as EMBOSS MATCHER, EMBOSS WATER, EMBOSS STRETCHER, EMBOSS NEEDLE, EMBOSS LALIGN, BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software.Those skilled in the art can determine the appropriate parameters for measuring the comparison, are included in any algorithm required for maximum comparison on the total length of the sequence being compared.

[0211] In the case of amino acid sequence comparisons using ALIGN-2, the % amino acid sequence identity of a given amino acid sequence A with, and, or against a given amino acid sequence B (which can alternatively be expressed as a given amino acid sequence A having or comprising a certain % amino acid sequence identity with, and, or against a given amino acid sequence B) is calculated as follows: 100 multiplied by a fraction X / Y, where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in the program's alignment of A and B, and where Y is the total number of amino acid residues in B. It will be understood that when the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not be equal to the % amino acid sequence identity of B to A. Unless specifically stated otherwise, all % amino acid sequence identity values ​​used herein are obtained using the ALIGN-2 computer program as in the preceding paragraph.

[0212] The terms "complementarity determining region" and "CDR", which are synonymous with "hypervariable region" or "HVR", are known in the art to refer to non-contiguous sequences of amino acids within an antibody variable region that confer antigen specificity and / or binding affinity. Typically, there are three CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region and three CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region. "Framework region" and "FR" are known in the art and refer to the non-CDR portions of the heavy and light chain variable regions. Typically, there are four FRs (FR-H1, FR-H2, FR-H3, and FR-H4) in each full-length heavy chain variable region and four FRs (FR-L1, FR-L2, FR-L3, and FR-L4) in each full-length light chain variable region.The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those described in Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding sitetopography," J. Mol. Biol. 262, 732-745. ("Contact" numbering scheme); Lefranc MP et al., "IMGTunique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003 Jan; 27 (1): 55-77 ("IMGT" numbering scheme); Honegger A and Plückthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8; 309 (3): 657-70, ("Aho" numbering scheme); and Whitelegg NR and Rees AR, "WAM: an improved algorithm for modelling antibodies on the WEB," Protein Eng. 2000 Dec; 13 (12): 819-24 ("AbM" numbering scheme). In certain embodiments, the CDRs of the antibodies herein can be defined by a method selected from Kabat, Chothia, IMGT, Aho, AbM, or a combination thereof.

[0213] The boundaries of a given CDR or FR can vary depending on the scheme used to identify them. For example, the Kabat scheme is based on structural alignments, while the Chothia scheme is based on structural information. The numbering of the Kabat and Chothia schemes is based on the most common antibody region sequence lengths, wherein insertions are accommodated by inserting letters, such as "30a," and deletions occur in some antibodies. These two schemes place certain insertions and deletions ("indels") in different positions, resulting in different numbering. The Contact scheme is based on the analysis of complex crystal structures, and is similar to the Chothia numbering scheme in many respects.

[0214] Isolated polypeptide or polypeptide complex composition

[0215] TROP2 binding domain and peptide shield (P1)

[0216] Disclosed herein are isolated polypeptides or polypeptide complexes comprising a recombinant antibody or antigen-binding fragment comprising a calcium signal transducer 2 (TROP2) binding domain that is selectively activated in a tumor microenvironment. In some embodiments, the TROP2 binding domain is linked to a peptide (P1) comprising an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-163. In some embodiments, P1 shields the TROP2 binding domain from binding to TROP2 in healthy tissue and is released from the TROP2 binding domain in the tumor microenvironment by a tumor-specific protease.

[0217] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2. NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YWX 14 X 15 X 16 X 17 wherein X8 is R, S, T, Q, D, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17is V, G, P, L, I, M, S, T, or A; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease.

[0218] Disclosed herein are isolated polypeptides or polypeptide complexes comprising the following Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2. NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YWX 14 X 15 X 16 X 17 wherein X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17 is V, G, P, L, I, M, S, T, or A; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease.

[0219] Disclosed herein are isolated polypeptides or polypeptide complexes comprising the following formula I: A1-L1-P1, wherein: A1 is a recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2. NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YW X 14 X 15 X 16 X 17 wherein X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17 is V, G, P, L, I, M, S, T or A; P1 is a peptide that binds to A1, wherein P1 is an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163; and L1 is a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease.

[0220] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 is a recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2. NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YW X 14 X 15 X 16 X 17wherein X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17 is V, G, P, L, I, M, S, T or A; P1 is a peptide that binds to A1, wherein P1 is an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163; and L1 is a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease.

[0221] Table 1. TROP2 binding domain light chain complementarity determining regions (CDRs) based on the IMGT CDR numbering system.

[0222] Construct description Amino acid sequence (N to C) SEQ ID NO: CDR1-L QDVSIA 1 CDR2-L SA 2 CDR3-L hRS7 QQHYITPLT 3 CDR3-L Q89A AQHYITPLT 4 CDR3-L Q90A QAHYITPLT 5 CDR3-L H91A QQAYITPLT 6 CDR3-L Y92A QQHAITPLT 7 CDR3-L I93A QQHYATPLT 8 CDR3-L T94A QQHYIAPLT 9 CDR3-L P95A QQHYITALT 10 CDR3-L L96A QQHYITPAT 11 CDR3-L T97A QQHYITPLA 12

[0223] Table 2. TROP2 binding domain heavy chain complementarity determining regions (CDRs) based on the IMGT CDR numbering system.

[0224] Construct description Amino acid sequence (N to C) SEQ ID NO: CDR1-H GYTFTNYG 13 CDR2-H INTYTGEP 14 CDR3-H hRS7 ARGGFGSSYWYFDV 15 CDR3-H R98A AAGGFGSSYWYFDV 16 CDR3-H G99A ARAGFGSSYWYFDV 17 CDR3-H G100A ARGAFGSSYWYFDV 18 CDR3-H F101A ARGGAGSSYWYFDV 19 CDR3-H G102A ARGGFASSYWYFDV 20 CDR3-H S103A ARGGFGASYWYFDV 21 CDR3-H S104A ARGGFGSAYWYFDV 22 CDR3-H Y105A ARGGFGSSAWYFDV 23 CDR3-H W106A ARGGFGSSYAYFDV 24 CDR3-H Y107A ARGGFGSSYWAFDV 25 CDR3-H F108A ARGGFGSSYWYADV 26 CDR3-H D109A ARGGFGSSYWYFAV 27 CDR3-H V110A ARGGFGSSYWYFDA 28

[0225] Table 3. Peptide mask sequences for TROP2 Fab alanine scanning sequences

[0226]

[0227]

[0228] In some embodiments, X1 is Q, N, D, E, or A; X2 is Q, N, D, E, or A; X3 is I, V, L, or A; X4 is T, S, or A; X5 is P, G, or A; X6 is L, V, I, or A; X7 is T, S, or A; X8 is R, K, or A; X9 is G, S, T, or A; X 10 Is F, Y or A; X 11 is G, S, T or A; X 12 is S, G, T or A; X 13is S, G, T or A; X 14 is Y, W, F or A; X 15 is F, Y, W or A; X 16 is D, E, Q, N, or A; and X 17 is V, L, I or A. In some embodiments, X1 is Q; and X6 is L. In some embodiments, X8 is R; X 10 It's F; X 11 It is G; X 14 It is Y; X 15 is F; and X 16 is D. In some embodiments, X1 is Q, S, T, D, N, E, or A; X2 is Q, S, T, D, N, E, or A; X3 is I, G166, P, V, L, M, or A; X4 is T, G, S, M, H, N, Q, or A; X5 is P, G, V, L, I, M, or A; X6 is L, G, P, V, I, M, or A; X7 is T, G, S, M, H, N, Q, or A; X8 is R, H, K, or A; X9 is G, P, V, L, I, M, S, T, or A; X 10 is F, Y, W, V, L, I, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q or A; X 13 is S, G, T, M, N, Q or A; X 14 is Y, F, W, V, L, I, or A; X 15 is F, Y, W, V, L, I, or A; X 16 is D, Q, N, E, S, T, or A; and X 17 is V, G, P, L, I, M, or A. In some embodiments, X1 is Q, N, or A; X2 is Q, N, or A; X3 is I, V, L, or A; X4 is T, S, or A; X5 is P, G, or A; X6 is L, V, I, or A; X7 is T, S, or A; X8 is R, K, or A; X9 is G, V, S, T, or A; X 10 Is F, Y or A; X 11 is G, V, S, T or A; X 12 is S, G, T or A; X 13 is S, G, T or A; X 14 is Y, W or A; X 15 Is F, Y or A; X 16 is D, E, or A; and X 17 is V, G, L, I or A. In some embodiments, X1 is Q; and X6 is L. In some embodiments, X8 is R; X 10 It's F; X 11 It is G; X 14 It is Y; X15 is F; and X 16 It’s D.

[0229] In some embodiments, CDR3-L comprises an amino acid selected from SEQ ID NOs: 3-5 and 8-12.

[0230] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 5, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2; CDR3-L: SEQ ID NO: 8, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; and CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 12 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15.

[0231] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 5, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 9, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 10, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 11, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15.In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 12 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15.

[0232] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 5, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 9, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15.

[0233] In some embodiments, CDR3-H comprises amino acids selected from SEQ ID NOs: 16-17, 19-22, and 25-28. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 16; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 17; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 25;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:27; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:28.;

[0234] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 16. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 17. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 19. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 20. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 21. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 22.In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 25. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 26. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR sequence set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28.

[0235] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 17; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 21; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: NO:22; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:28.

[0236] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 16, CDR2-L: SEQ ID NO: 17, CDR3-H: SEQ ID NO: 18; NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:27;and CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28, P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease.;

[0237] Disclosed herein are isolated polypeptides or polypeptide complexes comprising Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 16, CDR2-L: SEQ ID NO: 17, CDR3-H: SEQ ID NO: 18; NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2;CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2; CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14; CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28, P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163;and L1 comprises a linker portion that connects A1 to P1 and is a substrate for a tumor-specific protease.;

[0238] Disclosed herein are isolated polypeptides or polypeptide complexes comprising Formula I: A1-L1-P1, wherein: A1 is a recombinant antibody or an antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 16, CDR2-L: SEQ ID NO: 17, CDR3-H: SEQ ID NO: 18; NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2;CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2; CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14; CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28, P1 is a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163;and L1 is a linker moiety that connects A1 to P1 and is a substrate for a tumor-specific protease.;

[0239] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 is a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 16, CDR2-L: SEQ ID NO: 17, CDR3-H: SEQ ID NO: 18; NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2;CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2; CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14; CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28, P1 is a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163;and L1 is a linker moiety that connects A1 to P1 and is a substrate for a tumor-specific protease.;

[0240] In some embodiments, the TROP2 binding domain comprises a Fab, Fab', (Fab')2, or a single-chain variable fragment (scFv). In some embodiments, the TROP2 binding domain is a Fab. In some embodiments, the immunoglobulin light chain comprises the variable domain of an immunoglobulin kappa (IgK) or immunoglobulin lambda (IgL) light chain. In some embodiments, the immunoglobulin heavy chain comprises the variable domain of an IgG1, IgG2, IgG3, or IgG4 heavy chain.

[0241] In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NO: 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, or 73. In some embodiments, the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NO: 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, or 74.

[0242] In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 33, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 34. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 35, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 36. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 37, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 40. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 41, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 42. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:43, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:44.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 47, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 48.

[0243] In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 50. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 51, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 54. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 55, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 56. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 57, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 58. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:62.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 63, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 64. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 65, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 68. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74.

[0244] Table 4. TROP2 binding domain (TBD) heavy and light chain sequences

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 31, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 32.

[0252] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 5, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 33, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 34.

[0253] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 40.

[0254] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 9, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 41, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 42.

[0255] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 10 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 43, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 44.

[0256] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 11, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 45, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 46.

[0257] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 12, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 47, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 48.

[0258] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 16; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 50.

[0259] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 17; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 51, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 52.

[0260] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 19; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 55, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 56.

[0261] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and

[0262] CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 20; and the immunoglobulin light chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 57, and the immunoglobulin heavy chain comprises an amino acid sequence at least 90% identical to SEQ ID NO: 58.

[0263] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 21; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 59, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 60.

[0264] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 22; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 62.

[0265] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 25; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 67, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 68.

[0266] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 26; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 70.

[0267] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 72.

[0268] In some embodiments, the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the amino acid sequence comprises CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 28; and the immunoglobulin light chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 73, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 74.

[0269] In some embodiments, the TROP2 binding domain has a faster off-rate (greater k) for TROP2 binding than a TROP2 binding domain comprising an immunoglobulin light chain according to SEQ ID NO: 29 and an immunoglobulin heavy chain according to SEQ ID NO: 30, as measured under substantially similar kinetic assay conditions. dissIn some embodiments, the TROP2 binding domain has weaker binding to TROP2 as measured by an enzyme-linked immunosorbent assay (ELISA) under substantially similar assay conditions than a TROP2 binding domain comprising an immunoglobulin light chain according to SEQ ID NO: 29 and an immunoglobulin heavy chain according to SEQ ID NO: 30. In some embodiments, the TROP2 binding domain has an increased EC for TROP2 as measured by an ELISA under substantially similar assay conditions than a TROP2 binding domain comprising an immunoglobulin light chain according to SEQ ID NO: 29 and an immunoglobulin heavy chain according to SEQ ID NO: 30. 50 In some embodiments, TROP2 comprises the amino acid sequence according to SEQ ID NO: 396 (see Table 13).

[0270] In some embodiments, P1 weakens the binding of A1 to TROP2. In some embodiments, P1 binds to A1 through ionic interactions, electrostatic interactions, hydrophobic interactions, Pi stacking interactions, and H-bond interactions, or a combination thereof. In some embodiments, P1 binds to A1 at or near the antigen binding site. In some embodiments, when L1 is cleaved by a tumor-specific protease, P1 dissociates from A1, thereby exposing A1 to TROP2. In some embodiments, P1 has less than 75% sequence identity with TROP2. In some embodiments, P1 has less than 80% sequence identity with TROP2. In some embodiments, P1 has less than 85% sequence identity with TROP2. In some embodiments, P1 has less than 90% sequence identity with TROP2. In some embodiments, P1 has less than 95% sequence identity with TROP2. In some embodiments, P1 comprises a de novo generated amino acid sequence that shares less than 10% sequence identity with TROP2. In some embodiments, P1 comprises at least two cysteine ​​amino acid residues. In some embodiments, P1 comprises a cyclic peptide or a linear peptide. In some embodiments, P1 comprises a cyclic peptide. In some embodiments, P1 comprises a linear peptide. In some embodiments, P1 comprises a modified amino acid or an unnatural amino acid, or a modified unnatural amino acid, or a combination thereof. In some embodiments, P1 does not comprise albumin or an albumin fragment. In some embodiments, P1 does not comprise an albumin binding domain.

[0271] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 102, 107, 123, and 124, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 102, 107, 123, and 124. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and P1 comprises an amino acid sequence according to SEQ ID NO: 102 or SEQ ID NO: 107, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 102 or SEQ ID NO: 107. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 29, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 30, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 102, 107, 123, and 124, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 102, 107, 123, and 124.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:29, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:30, and P1 comprises an amino acid sequence according to SEQ ID NO:102 or SEQ ID NO:107, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO:102 or SEQ ID NO:107.

[0272] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 40, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 40, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142, and 150.

[0273] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 16, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 16, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 50, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 50, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 142, and 150.

[0274] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 22, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 22, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 141, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 62, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:62, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs:107, 141, 142, and 150.

[0275] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 26, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 26, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 141, 142, and 150. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 70, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 70, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 141, 142, and 150.

[0276] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 27, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 27, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, 158, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, 158. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:72, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:72, and P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 109, 116, 141, 142, 148, 149, 150, and 158, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs:107, 109, 116, 141, 142, 148, 149, 150, and 158.

[0277] CD3 binding domain

[0278] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain. In some embodiments, the isolated polypeptide or polypeptide complex is according to the following formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking portion that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the isolated polypeptide or polypeptide complex comprises the following formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking portion that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the isolated polypeptide or polypeptide complex comprises the following formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 is a CD3 binding domain, P2 is a peptide that binds to B2, and L2 is a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the isolated polypeptide or polypeptide complex is according to the following formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 is a CD3 binding domain, P2 is a peptide that binds to B2, and L2 is a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease.

[0279] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, CDR1-L comprising the amino acid sequence of SEQ ID NO: 75; CDR2-L comprising the amino acid sequence of SEQ ID NO: 76; CDR3-L comprising the amino acid sequence of SEQ ID NO: 77; CDR1-H comprising the amino acid sequence of SEQ ID NO: 78; and CDR2-H comprising the amino acid sequence of SEQ ID NO: 79; and CDR3-H comprising the amino acid sequence of SEQ ID NO: 80.

[0280] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H; wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75 or SEQ ID NO: 259; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: NO:76 amino acid sequence; wherein CDR3-L comprises the amino acid sequence of Z1-Z2-W-Z3-Z4-Z5-Z6-W-Z7-Z8; wherein Z1 is V, G, P, L, I, M, S, T or A; Z2 is L, G, P, V, I, M, S, T or A; Z3 is Y, F, W, V, L, I, G or A; Z4 is S, G, T, M, N, Q, H or A; Z5 is N, Q, S, T, D, E, H, K, R or A; Z6 is R, S, T, Q, D, E, H, K, N or A; Z7 is V, G, P, L, I, M, S, T or A; and Z8 is F, Y, W, V, L, I, G or A; wherein CDR1-H comprises SEQ ID NO:78 or SEQ ID NO: NO:270 amino acid sequence; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:79; and wherein CDR3-H comprises Z9-Z 10 -Z 11 -Z 12 -NZ 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -YZ 20 -AZ 21 wherein Z9 is V, G, P, L, I, M, S, T or A; Z10 is R, S, T, Q, D, E, H, K, N or A; Z 11 is H, R, K, G, T, S, N, Q or A; Z 12 is G, P, V, L, I, M, S, T, or A; Z 13 is F, Y, W, V, L, I, G, or A; Z 14 is G, P, V, L, I, M, S, T, or A; Z 15 is N, Q, S, T, D, E, H, K, R, or A; Z 16 is S, G, T, M, N, Q, H or A; Z 17 Y, F, W, V, L, I, G, or A; Z 18 is I, G, P, V, L, M, S, T, or A; Z 19 is S, G, T, M, N, Q, H or A; Z 20 is W, F, Y, V, L, I, G, or A; and Z 21 is Y, F, W, V, L, I, G, or A. In some embodiments, Z1 is V, G, L, I, or A; Z2 is L, V, I, or A; Z3 is Y, W, F, or A; Z4 is S, G, T, or A; Z5 is N, Q, D, E, or A; Z6 is R, K, or A; Z7 is V, G, L, I, or A; Z8 is F, Y, W, or A; Z9 is V, G, L, I, or A; Z 10 R, K or A; Z 12 is G, S, T or A; Z 13 Is F, Y, W or A; Z 14 is G, S, T or A; Z 15 is N, Q, D, E or A; Z 16 Is S, G, T or A; Z 17 Y, W, F or A; Z 18 Is I, V, L or A; Z 19 Is S, G, T or A; Z 20 is W, Y, F, or A; and Z 21 is Y, W, F or A. In some embodiments, Z8 is F. In some embodiments, Z 10 It is R; Z 11 It is H; Z 13 It's F; Z 18 It is I; Z 19 is S; and Z 20 It’s W.

[0281] Table 5. CD3 binding domain light chain complementarity determining regions (CDRs) based on the IMGT CDR numbering system.

[0282] Construct description Amino acid sequence (N to C) SEQ ID NO: CDR1-L TGAVTSGNY 75 CDR2-L GTK 76 CDR3-L VLWYSNRWV 77 CDR1-L TGAVTSANY 259 CDR3-L V231A ALWYSNRWV 260 CDR3-L L232A VAWYSNRWV 261 CDR3-L W233A VLAYSNRWV 262 CDR3-L Y234A VLWASNRWV 263 CDR3-L S235A VLWYANRWV 264 CDR3-L N236A VLWYSARWV 265 CDR3-L R237A VLWYSNAWV 266 CDR3-L W238A VLWYSNRAV 267 CDR3-L V239A VLWYSNRWA 268 CDR3-L F240A VLWYSNRWVA 269

[0283] Table 6. CD3 binding domain heavy chain complementarity determining regions (CDRs) based on the IMGT CDR numbering system.

[0284] Construct description Amino acid sequence (N to C) SEQ ID NO: CDR1-H GFTFNKYA 78 CDR2-H IRSKYNNYAT 79 CDR3-H VRHGNFGNSYISYWAY 80 CDR1-H GFTFQKYA 270 CDR3-H V99A ARHGNFGNSYISYWAY 271 CDR3-H R100A VAHGNFGNSYISYWAY 272 CDR3-H H101A VRAGNFGNSYISYWAY 273 CDR3-H G102A VRHANFGNSYISYWAY 274 CDR3-H N103A VRHGAFGNSYISYWAY 275 CDR3-H F104A VRHGNAGNSYISYWAY 276 CDR3-H G105A VRHGNFANSYISYWAY 277 CDR3-H N106A VRHGNFGASYISYWAY 278 CDR3-H S107A VRHGNFGNAYISYWAY 279 CDR3-H Y108A VRHGNFGNSAISYWAY 280 CDR3-H I109A VRHGNFGNSYASYWAY 281 CDR3-H S110A VRHGNFGNSYIAYWAY 282 CDR3-H Y111A VRHGNFGNSYISAWAY 283 CDR3-H W112A VRHGNFGNSYISYAAY 284 CDR3-H Y114A VRHGNFGNSYISYWAA 285 CDR3-H I109V VRHGNFGNSYVSYWAY 286

[0285] In some embodiments, the CDR3-L of the CD3 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 77, 260-261, 263-266, and 268-269. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 263, and CDR1-H: SEQ ID NO: NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQID NO:76, CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:268 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80;and CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:269 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 263, and CDR1-H: SEQ ID NO: NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQID NO:76, CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80;and CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 268 and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80. ;

[0286] In some embodiments, CDR3-H of the CD3 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 80, 271-274, 276-282, and 284-285. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 271; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 272; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 272. NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:273; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:274; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:276; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:281; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:282; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:284; and CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:285.;

[0287] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 271; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 274; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 274. NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; and CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:285.

[0288] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 80; CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 81. ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:262 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:263 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:267, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:268, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:269, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:271; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77, CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:272; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:273; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:274; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:275;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:276; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:281; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:282; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:283;CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:284; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:285; and CDR1-L: SEQ ID NO:259, CDR2-L: SEQ ID NO:76, CDR3-L: SEQ ID NO:260 and CDR1-H: SEQ ID NO:270, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:286. ;

[0289] In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises the variable domain of an immunoglobulin kappa (IgK) or immunoglobulin lambda (IgL) light chain. In some embodiments, the immunoglobulin heavy chain of the CD3 binding domain comprises the variable domain of an IgG1, IgG2, IgG3, or IgG4 heavy chain. In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 81. In some embodiments, the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 82. In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:81, and the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:82.

[0290] In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 99. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 303. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 304. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 306. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 307. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 309. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 310. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 311. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 312. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 313. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:314.In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 315. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 316. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 317. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 318. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 319. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 320. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 321. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 322. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 323. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 324. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 325. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 326. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:327.In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 328. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329.

[0291] Table 7. CD3 binding domain heavy and light chain sequences

[0292]

[0293]

[0294]

[0295]

[0296]

[0297] In some embodiments, the CD3 binding domain comprises Fab, Fab', (Fab')2 or a single-chain variable fragment (scFv). In some embodiments, the CD3 binding domain is a scFv. In some embodiments, the N-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the C-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the N-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the N-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the C-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the N-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin heavy chain of the TROP2 binding domain binds to the N-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the N-terminus of the immunoglobulin light chain of the TROP2 binding domain binds to the C-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin light chain of the TROP2 binding domain is bound to the N-terminus of the immunoglobulin light chain of the CD3 binding domain. In some embodiments, the N-terminus of the immunoglobulin light chain of the TROP2 binding domain is bound to the C-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the C-terminus of the immunoglobulin light chain of the TROP2 binding domain is bound to the N-terminus of the immunoglobulin heavy chain of the CD3 binding domain. In some embodiments, the CD3 binding domain is an scFv and the TROP2 binding domain is a Fab or Fab'. In some embodiments, the scFv is bound to the immunoglobulin heavy chain of Fab or Fab'. In some embodiments, the scFv is bound to the immunoglobulin light chain of Fab or Fab'. In some embodiments, the immunoglobulin light chain of the scFv is bound to the immunoglobulin heavy chain of Fab or Fab'. In some embodiments, the immunoglobulin light chain of the scFv is bound to the immunoglobulin light chain of Fab or Fab'. In some embodiments, the immunoglobulin light chain of the scFv is bound to the immunoglobulin heavy chain of Fab or Fab'. In some embodiments, the immunoglobulin heavy chain of the scFv is associated with the immunoglobulin heavy chain of the Fab or Fab'.In some embodiments, the immunoglobulin heavy chain of the scFv is associated with the immunoglobulin light chain of the Fab or Fab'.

[0298] T cell engager (TCE) compositions

[0299] In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and SEQ ID NO: 88. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 89 and SEQ ID NO: 90. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 91 and SEQ ID NO: 92. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 93 and SEQ ID NO: 94. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95 and SEQ ID NO: 96. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 97 and SEQ ID NO: 98.

[0300] In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 330 and SEQ ID NO: 331. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 332 and SEQ ID NO: 333. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 332 and SEQ ID NO: 333. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 334 and SEQ ID NO: 335. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 336 and SEQ ID NO: 337. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 338 and SEQ ID NO: 339. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 340 and SEQ ID NO: 341. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 342 and SEQ ID NO: 343. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 344 and SEQ ID NO: 345. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:346 and SEQ ID NO:347.In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 348 and SEQ ID NO: 349. In some embodiments, the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 350 and SEQ ID NO: 351.

[0301] Table 8. TROP2-T cell engager (TCE) sequences

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318]

[0319]

[0320]

[0321]

[0322]

[0323] In some embodiments, the recombinant antibody or antigen-binding fragment thereof has weaker cytotoxic activity than a recombinant antibody or antigen-binding fragment thereof comprising an immunoglobulin light chain according to SEQ ID NO: 83 or 85 and an immunoglobulin heavy chain according to SEQ ID NO: 84 or 86, as measured in an in vitro tumor cell killing assay under substantially similar assay conditions.

[0324] CD3 binding domain peptide shield (P2)

[0325] In some embodiments, P2 weakens the binding of B2 to CD3. In some embodiments, P2 binds to B2 through ionic interactions, electrostatic interactions, hydrophobic interactions, Pi stacking interactions, and H-bond interactions, or a combination thereof. In some embodiments, P2 binds to B2 at or near the antigen binding site. In some embodiments, when L2 is cleaved by a tumor-specific protease, P2 dissociates from B2, thereby exposing B2 to CD3. In some embodiments, P2 has less than 70% sequence identity with CD3. In some embodiments, P2 has less than 75% sequence identity with CD3. In some embodiments, P2 has less than 80% sequence identity with CD3. In some embodiments, P2 has less than 85% sequence identity with CD3. In some embodiments, P2 has less than 90% sequence identity with CD3. In some embodiments, P2 has less than 95% sequence identity with CD3. In some embodiments, P2 comprises a de novo generated amino acid sequence that shares less than 10% sequence identity with CD3. In some embodiments, P2 comprises a peptide sequence of at least 5 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 6 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 10 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 10 amino acids in length and no more than 20 amino acids in length. In some embodiments, P2 comprises a peptide sequence of at least 16 amino acids in length. In some embodiments, P2 comprises a peptide sequence of no more than 40 amino acids in length. In some embodiments, P2 comprises at least two cysteine ​​amino acid residues. In some embodiments, P2 comprises a cyclic peptide or a linear peptide. In some embodiments, P2 comprises a cyclic peptide. In some embodiments, P2 comprises a linear peptide. In some embodiments, P2 comprises a modified amino acid or a non-natural amino acid, or a modified non-natural amino acid, or a combination thereof. In some embodiments, P2 does not comprise albumin or an albumin fragment. In some embodiments, P2 does not comprise an albumin binding domain. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO: 292. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 292.

[0326] In some embodiments, P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 287-302, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 287-302. In some embodiments, P2 comprises an amino acid sequence according to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 273, and P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 273, and P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 273, and P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305, and P2 comprises the amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298.In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305, and P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence according to SEQ ID NO: 305, and P2 comprises an amino acid sequence according to SEQ ID NO: 295.

[0327] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 276, and P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 77, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 276, and P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308, and P2 comprises the amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence according to SEQ ID NO: 308, and P2 comprises an amino acid sequence according to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 259, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, CDR1-H: SEQ ID NO: 270, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 286, and P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298.In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 259, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, CDR1-H: SEQ ID NO: 270, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 286, and P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 259, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 260, CDR1-H: SEQ ID NO: 270, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 286, and P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329, and P2 comprises the amino acid sequence according to any one of SEQ ID NOs: 292, 295, and 298, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 292, 295, and 298. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329, and P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence according to SEQ ID NO: 329, and P2 comprises an amino acid sequence according to SEQ ID NO: 295.

[0328] In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 80, and P2 comprises an amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the immunoglobulin light chain and the immunoglobulin heavy chain of the CD3 binding domain comprise a CDR set comprising CDR1-L: SEQ ID NO: 75, CDR2-L: SEQ ID NO: 76, CDR3-L: SEQ ID NO: 261, CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, and CDR3-H: SEQ ID NO: 80, and P2 comprises the amino acid sequence according to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 319, and P2 comprises the amino acid sequence according to SEQ ID NO: 295, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 295. In some embodiments, the CD3 binding domain comprises the amino acid sequence according to SEQ ID NO:319, and P2 comprises the amino acid sequence according to SEQ ID NO:295.

[0329] Table 9. Peptide masks of CD3 scFv sequences

[0330] peptides Amino acid sequence (N to C) SEQ ID NO: Peptide-65 QGCGTIADPEPHCW 287 Peptide-66 GSQCLGPEWEVCPY 288 Peptide-67 VYCGPEFDESVGCM 289 Peptide-68 DWCWYSPDNDGLCD 290 Peptide-69 SECLVDLQSDPCYW 291 Peptide-70 YLCGPDGDETLACY 292 Peptide-71 VDCGPDGDESILCY 293 Peptide-72 AYCGPEFDESVGCM 294 Peptide-73 VYCGPEFDESVGCA 295 Peptide-74 ASQCLGPEWEVCPY 296 Peptide-75 GSACLGPEWEVCPY 297 Peptide-76 YLCGPDADETLACY 298 Peptide-77 YLCGPDTDETLACY 299 Peptide-78 IDFCAVYKWPVCQV 300 Peptide-79 IDFCLIYNWPVCDT 301 Peptide-80 PDFCAVYRWPICYQ 302

[0331] Connecting part (L1 or L2)

[0332] In some embodiments, L1 or L2 is a peptide sequence having at least 5 to no more than 50 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 10 to no more than 30 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 10 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 18 amino acids. In some embodiments, L1 or L2 is a peptide sequence having at least 26 amino acids. In some embodiments, L1 or L2 comprises (G2S) nwherein n is an integer from 1 to 3. In some embodiments, L1 or L2 comprises (G2S) n wherein n is an integer of at least 1. In some embodiments, L1 or L2 comprises a compound selected from (G2S) n , (GS) n 、(GSGGS) n 、(GGGS) n 、(GGGGS) n and (GSSGGS) n wherein n is an integer of at least 1. In some embodiments, the tumor-specific protease is selected from a metalloprotease, a serine protease, a cysteine ​​protease, a threonine protease, and an aspartic protease. In some embodiments, L1 or L2 comprises an amino acid sequence cleavable by urokinase, an amino acid sequence cleavable by a membrane-type serine protease (matriptase), or an amino acid sequence cleavable by a matrix metalloprotease. In some embodiments, L1 or L2 comprises a sequence according to any one of SEQ ID NOs: 226-254. In some embodiments, L1 is bound to the N-terminus of A1. In some embodiments, L1 is bound to the C-terminus of A1. In some embodiments, L2 is bound to the N-terminus of B2. In some embodiments, L2 is bound to the C-terminus of B2.

[0333] Table 10. L1 or L2

[0334]

[0335] Half-life extending molecules (H1)

[0336] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a half-life extending molecule (H1). In some embodiments, H1 is linked to P1. In some embodiments, H1 is linked to P2. In some embodiments, H1 does not block the CD3 binding domain from CD3. In some embodiments, the half-life extending molecule (H1) does not have binding affinity for CD3. In some embodiments, the half-life extending molecule (H1) does not shield the isolated polypeptide or polypeptide complex from CD3. In some embodiments, H1 comprises a sequence according to SEQ ID NOs: 255-258. In some embodiments, H1 comprises an amino acid sequence having a repeating sequence motif. In some embodiments, H1 comprises an amino acid sequence having a highly ordered secondary structure. In some embodiments, H1 comprises a polymer. In some embodiments, the polymer is polyethylene glycol (PEG). In some embodiments, H1 comprises albumin. In some embodiments, H1 comprises an Fc domain. In some embodiments, the albumin is serum albumin. In some embodiments, the albumin is human serum albumin. In some embodiments, H1 comprises a polypeptide, a ligand, or a small molecule. In some embodiments, the polypeptide, ligand, or small molecule binds to a serum protein or fragment thereof, a circulating immunoglobulin or fragment thereof, or CD35 / CR1. In some embodiments, the serum protein comprises thyroxine-binding protein, transthyretin, 1-acid glycoprotein, transferrin, transferrin receptor or a transferrin-binding portion thereof, fibrinogen, or albumin. In some embodiments, the circulating immunoglobulin molecule comprises IgG1, IgG2, IgG3, IgG4, slgA, IgM, or IgD. In some embodiments, the serum protein is albumin. In some embodiments, the polypeptide is an antibody. In some embodiments, the antibody comprises a single domain antibody, a single chain variable fragment, or a Fab. In some embodiments, the single domain antibody comprises a single domain antibody that binds to albumin. In some embodiments, the single domain antibody is a human antibody or a humanized antibody. In some embodiments, the single domain antibody is selected from 645gH1gL1, 645dsgH5gL4, 23-13-A01-sc02, A10m3 or a fragment thereof, DOM7r-31, DOM7h-11-15, Alb-1, Alb-8, Alb-23, 10G, 10E, and SA21.In some embodiments, the single-domain antibody comprises a complementarity determining region (CDR): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, HC-CDR2, and HC-CDR3 of the single-domain antibody comprise: HC-CDR1: SEQ ID NO: 255, HC-CDR2: SEQ ID NO: 256, and HC-CDR3: SEQ ID NO: 257; and wherein the CDRs comprise 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2, or HC-CDR3. In some embodiments, H1 comprises an amino acid sequence according to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 258. In some embodiments, H1 comprises a modified amino acid or a non-natural amino acid, or a modified non-natural amino acid, or a combination thereof. In some embodiments, the modified amino acid or the modified non-natural amino acid comprises a post-translational modification. In some embodiments, H1 comprises a linking portion (L3) connecting H1 to P1 or P2. In some embodiments, L3 is a peptide sequence having at least 5 to no more than 50 amino acids. In some embodiments, L3 is a peptide sequence having at least 10 to no more than 30 amino acids. In some embodiments, L3 is a peptide sequence having at least 10 amino acids. In some embodiments, L3 is a peptide sequence having at least 18 amino acids. In some embodiments, L3 is a peptide sequence having at least 26 amino acids. In some embodiments, L3 comprises a peptide selected from (G2S). n , (GS) n 、(GSGGS) n 、(GGGS) n 、(GGGGS) n and (GSSGGS) n wherein n is an integer of at least 1. In some embodiments, L3 comprises a sequence according to any one of SEQ ID NOs: 226-254.

[0337] Table 11. H1 sequences

[0338]

[0339] Peptide complex

[0340] In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 166 and SEQ ID NO: 167. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 168 and SEQ ID NO: 169. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 170 and SEQ ID NO: 171. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 172 and SEQ ID NO: 173. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 174 and SEQ ID NO: 175. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 176 and SEQ ID NO: 177. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 178 and SEQ ID NO: 179. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 180 and SEQ ID NO: 181. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 182 and SEQ ID NO: 183. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 184 and SEQ ID NO: 185.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 186 and SEQ ID NO: 187. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 188 and SEQ ID NO: 189. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 190 and SEQ ID NO: 191. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 192 and SEQ ID NO: 193. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 194 and SEQ ID NO: 195. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 196 and SEQ ID NO: 197. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 198 and SEQ ID NO: 199. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 200 and SEQ ID NO: 201. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 202 and SEQ ID NO: 203. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:204 and SEQ ID NO:205.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 206 and SEQ ID NO: 207. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 208 and SEQ ID NO: 209. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 210 and SEQ ID NO: 211. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 212 and SEQ ID NO: 213. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 214 and SEQ ID NO: 215. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 216 and SEQ ID NO: 217. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 218 and SEQ ID NO: 219. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 220 and SEQ ID NO: 221. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 222 and SEQ ID NO: 223. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:224 and SEQ ID NO:225.

[0341] In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 352 and SEQ ID NO: 353. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 354 and SEQ ID NO: 355. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 356 and SEQ ID NO: 357. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 358 and SEQ ID NO: 359. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 360 and SEQ ID NO: 361. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 362 and SEQ ID NO: 363. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 364 and SEQ ID NO: 365. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 366 and SEQ ID NO: 367. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 368 and SEQ ID NO: 369. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:370 and SEQ ID NO:371.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 372 and SEQ ID NO: 373. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 374 and SEQ ID NO: 375. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 376 and SEQ ID NO: 377. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 378 and SEQ ID NO: 379. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 380 and SEQ ID NO: 381. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 382 and SEQ ID NO: 383. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 384 and SEQ ID NO: 385. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 386 and SEQ ID NO: 387. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 388 and SEQ ID NO: 389. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:390 and SEQ ID NO:391.In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 392 and SEQ ID NO: 393. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 394 and SEQ ID NO: 395.

[0342] Table 12. Polypeptide complex sequences (masked sequences binding to TROP2 and CD3e)

[0343]

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361]

[0362]

[0363]

[0364]

[0365]

[0366]

[0367] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; wherein CDR3-L comprises the amino acid sequence of SEQ ID NO: 3; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and wherein CDR3-H comprises the amino acid sequence of SEQ ID NO: 15. NO:26 amino acid sequence; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119, and 123-163; and L1 comprises a linker that connects A1 to P1 and is a substrate for a tumor-specific protease. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 141, 142, and 150, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs:107, 141, 142, and 150. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142, and 150. In some embodiments, P1 comprises an amino acid sequence according to SEQ ID NO: 142, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 142. In some embodiments, P1 comprises an amino acid sequence according to SEQ ID NO: 142.

[0368] In some embodiments, the TROP2 binding domain comprises Fab, Fab', (Fab')2 or a single-chain variable fragment (scFv). In some embodiments, the TROP2 binding domain is Fab. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 92% identity to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 92% identity to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 70. In some embodiments, the immunoglobulin light chain comprises the amino acid sequence of SEQ ID NO: 69, and the immunoglobulin heavy chain comprises the amino acid sequence of SEQ ID NO: 70.

[0369] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain. In some embodiments, the isolated polypeptide or polypeptide complex is according to the formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 80.

[0370] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 276. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 259, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 260, CDR1-H comprises the amino acid sequence of SEQ ID NO: 270, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 286. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 273.

[0371] In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 81, and the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 82. In some embodiments, the CD3 binding domain comprises a Fab, Fab', (Fab')2, or a single-chain variable fragment (scFv). In some embodiments, the CD3 binding domain is an scFv. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 99. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 305.

[0372] In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO: 292. In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 295. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 200 and SEQ ID NO: 201. In some embodiments, the isolated polypeptide or polypeptide complex comprises the amino acid sequence of SEQ ID NO: 200 and SEQ ID NO: 201. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 362 and SEQ ID NO: 363. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 382 and SEQ ID NO: 383. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 390 and SEQ ID NO: 391.

[0373] Disclosed herein are isolated polypeptides or polypeptide complexes according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; wherein CDR3-L comprises the amino acid sequence of SEQ ID NO: 3; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and wherein CDR3-H comprises the amino acid sequence of SEQ ID NO: 15. NO:27; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163; and L1 comprises a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163. In some embodiments, P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, and 158. In some embodiments, P1 comprises an amino acid sequence according to SEQ ID NO: 107, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to SEQ ID NO: 107.

[0374] In some embodiments, the TROP2 binding domain comprises Fab, Fab', (Fab')2 or a single-chain variable fragment (scFv). In some embodiments, the TROP2 binding domain is Fab. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence having at least 92% identity to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence having at least 92% identity to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 72. In some embodiments, the immunoglobulin light chain comprises the amino acid sequence of SEQ ID NO: 71, and the immunoglobulin heavy chain comprises the amino acid sequence of SEQ ID NO: 72.

[0375] In some embodiments, the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain. In some embodiments, the isolated polypeptide or polypeptide complex is according to the formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises a CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking moiety that connects B2 to P2 and is a substrate for a tumor-specific protease. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 80.

[0376] In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 75, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 77, CDR1-H comprises the amino acid sequence of SEQ ID NO: 78, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 276. In some embodiments, the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising the complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, and the immunoglobulin heavy chain comprising the CDRs: CDR1-H, CDR2-H, and CDR3-H, and wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 259, CDR2-L comprises the amino acid sequence of SEQ ID NO: 76, CDR3-L comprises the amino acid sequence of SEQ ID NO: 260, CDR1-H comprises the amino acid sequence of SEQ ID NO: 270, CDR2-H comprises the amino acid sequence of SEQ ID NO: 79, and CDR3-L comprises the amino acid sequence of SEQ ID NO: 286.

[0377] In some embodiments, the immunoglobulin light chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 81, and the immunoglobulin heavy chain of the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 82. In some embodiments, the CD3 binding domain comprises a Fab, Fab', (Fab')2, or a single-chain variable fragment (scFv). In some embodiments, the CD3 binding domain is an scFv. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 99. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 308. In some embodiments, the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 329.

[0378] In some embodiments, P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO: 292. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 180 and SEQ ID NO: 181. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 352 and SEQ ID NO: 353. In some embodiments, the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 354 and SEQ ID NO: 355.

[0379] Polynucleotide encoding a recombinantly isolated polypeptide or polypeptide complex composition

[0380] In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding an isolated polypeptide or polypeptide complex according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2. NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, MS, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YW X 14 X 15 X 16 X 17 wherein X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17is V, G, P, L, I, M, S, T, or A; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linking portion that connects A1 to P1 and is a substrate for a tumor-specific protease.

[0381] In some embodiments, disclosed herein is an isolated recombinant nucleic acid molecule encoding an isolated polypeptide or polypeptide complex according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 4, and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15 NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQID NO:2, CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15;CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:10, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:11, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:12, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3, and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2;CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2; CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQID NO:14; CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:27; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2, CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:28, P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:100-163;and L1 comprises a linker portion that connects A1 to P1 and is a substrate for a tumor-specific protease.;

[0382] Pharmaceutical composition

[0383] In some embodiments, disclosed herein are pharmaceutical compositions comprising: (a) an isolated polypeptide or polypeptide complex according to any one of the embodiments disclosed herein and (b) a pharmaceutically acceptable excipient.

[0384] In some embodiments, the isolated polypeptide or polypeptide complex is according to Formula I: A1-L1-P1, wherein: A1 comprises a recombinant antibody or antigen-binding fragment thereof, the recombinant antibody or antigen-binding fragment thereof comprising a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L, and CDR3-L, the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H, and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; NO:2 amino acid sequence; and wherein CDR3-L comprises the amino acid sequence of X1X2HYX3X4X5X6X7; wherein X1 is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R or A; X3 is I, G, P, V, L, M, S, T or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T or A; X6 is L, G, P, V, I, M, S, T or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:14; and wherein CDR3-H comprises AX8X9GX 10 X 11 X 12 X 13 YWX 14 X 15 X 16 X 17 wherein X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G, or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17 is V, G, P, L, I, M, S, T, or A; P1 comprises a peptide bound to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or d...

Claims

1. An isolated polypeptide or polypeptide complex according to formula I: A1-L1-P1 (Formula I) in: A1 comprises a recombinant antibody or an antigen-binding fragment thereof, wherein the recombinant antibody or the antigen-binding fragment thereof comprises a tumor-associated calcium signal transducer 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, wherein the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO:2(SA); and wherein CDR3-L comprises an amino acid sequence of X1X2HYX3X4X5X6X7; Where X is Q, S, T, D, N, E, H, K, R or A; X2 is Q, S, T, D, N, E, H, K, R, or A; X3 is I, G, P, V, L, M, S, T, or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, S, T, or A; X6 is L, G, P, V, I, M, S, T, or A; and X7 is T, G, S, M, H, N, Q or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and Where CDR3-H contains AX8X9GX 10 X 11 X 12 X 13 YW 14 X 15 X 16 X 17 The amino acid sequence of Where X8 is R, S, T, Q, D, E, H, K, N or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I, G, or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q, H or A; X 13 is S, G, T, M, N, Q, H or A; X 14 is Y, F, W, V, L, I, G or A; X 15 is F, Y, W, V, L, I, G, or A; X 16 is D, Q, N, E, S, T, H, K, R, or A; and X 17 is V, G, P, L, I, M, S, T, or A; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linking moiety that connects A1 to P1 and is a substrate for tumor-specific proteases.

2. The isolated polypeptide or polypeptide complex according to claim 1, wherein X1 is Q, N, D, E or A; X2 is Q, N, D, E or A; X3 is I, V, L, or A; X4 is T, S or A; X5 is P, G or A; X6 is L, V, I or A; X7 is T, S or A; X8 is R, K or A; X9 is G, S, T or A; X 10 is F, Y or A; X 11 is G, S, T or A; X 12 is S, G, T or A; X 13 is S, G, T or A; X 14 is Y, W, F or A; X 15 is F, Y, W or A; X 16 is D, E, Q, N, or A; and X 17 It is V, L, I or A.

3. The isolated polypeptide or polypeptide complex according to claim 1, wherein X1 is Q; and X6 is L.

4. The isolated polypeptide or polypeptide complex according to claim 1, wherein X8 is R; X 10 It is F; X 11 It is G; X 14 It is Y; X 15 is F; and X 16 It is D.

5. The isolated polypeptide or polypeptide complex according to claim 1, wherein X1 is Q, S, T, D, N, E or A; X2 is Q, S, T, D, N, E, or A; X3 is I, G, P, V, L, M, or A; X4 is T, G, S, M, H, N, Q or A; X5 is P, G, V, L, I, M, or A; X6 is L, G, P, V, I, M, or A; X7 is T, G, S, M, H, N, Q or A; X8 is R, H, K or A; X9 is G, P, V, L, I, M, S, T or A; X 10 is F, Y, W, V, L, I or A; X 11 is G, P, V, L, I, M, S, T, or A; X 12 is S, G, T, M, N, Q or A; X 13 is S, G, T, M, N, Q or A; X 14 is Y, F, W, V, L, I or A; X 15 is F, Y, W, V, L, I or A; X 16 is D, Q, N, E, S, T, or A; and X 17 It is V, G, P, L, I, M or A.

6. The isolated polypeptide or polypeptide complex according to claim 5, wherein X1 is Q, N or A; X2 is Q, N or A; X3 is I, V, L, or A; X4 is T, S or A; X5 is P, G or A; X6 is L, V, I or A; X7 is T, S or A; X8 is R, K or A; X9 is G, V, S, T or A; X 10 is F, Y or A; X 11 is G, V, S, T or A; X 12 is S, G, T or A; X 13 is S, G, T or A; X 14 is Y, W or A; X 15 is F, Y or A; X 16 is D, E or A; and X 17 It is V, G, L, I or A.

7. The isolated polypeptide or polypeptide complex according to claim 5, wherein X1 is Q; and X6 is L.

8. The isolated polypeptide or polypeptide complex according to claim 7, wherein X8 is R; X 10 It is F; X 11 It is G; X 14 It is Y; X 15 is F; and X 16 It is D.

9. The isolated polypeptide or polypeptide complex according to claim 1, wherein CDR3-L comprises amino acids selected from the group consisting of SEQ ID NOs: 3-5 and 8-12.

10. The isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:4 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:10 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:11 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 15; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:12 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:

15.

11. The isolated polypeptide or polypeptide complex according to claim 1, wherein CDR3-H comprises amino acids selected from the group consisting of SEQ ID NOs: 16-17, 19-22 and 25-28.

12. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14; CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:

28.

13. An isolated polypeptide or polypeptide complex according to formula I: A1-L1-P1 (Formula I) in: A1 comprises a recombinant antibody or an antigen-binding fragment thereof, wherein the recombinant antibody or the antigen-binding fragment thereof comprises a tumor-associated calcium signaling protein 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, wherein the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the following: CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:4 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:5 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:6 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:7 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:8 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:9 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:10 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:11 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:12 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:15; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:16; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:17; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:18; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:19; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:20; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA); CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:21; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA); CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14; CDR3-H: SEQ ID NO:22; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:23; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:24; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:25; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:26; CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, CDR3-H: SEQ ID NO: 27; and CDR1-L: SEQ ID NO:1, CDR2-L: SEQ ID NO:2(SA), CDR3-L: SEQ ID NO:3 and CDR1-H: SEQ ID NO:13, CDR2-H: SEQ ID NO:14, CDR3-H: SEQ ID NO:28; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-163; and L1 comprises a linking moiety that connects A1 to P1 and is a substrate for tumor-specific proteases. 14 . The isolated polypeptide or polypeptide complex according to claim 1 , wherein the TROP2 binding domain comprises Fab, Fab′, (Fab′) 2 or a single chain variable fragment (scFv).

15. The isolated polypeptide or polypeptide complex according to claim 1, wherein the TROP2 binding domain is Fab.

16. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises a variable domain of an immunoglobulin kappa (IgK) or immunoglobulin lambda (IgL) light chain.

17. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin heavy chain comprises the variable domain of an IgG1, IgG2, IgG3 or IgG4 heavy chain.

18. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71 or 73.

19. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO:32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72 or 74.

20. An isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:

70.

21. An isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:

72.

22. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 102, 107, 123, and 124, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 102, 107, 123, and 124.

23. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to SEQ ID NO: 102 or SEQ ID NO: 107, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to SEQ ID NO: 102 or SEQ ID NO:

107.

24. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 29, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 30, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 102, 107, 123 and 124, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 102, 107, 123 and 124.

25. An isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:29, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:30, and wherein P1 comprises an amino acid sequence according to SEQ ID NO:102 or SEQ ID NO:107, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to SEQ ID NO:102 or SEQ ID NO:

107.

26. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.

27. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 8, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14 and CDR3-H: SEQ ID NO: 15, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142 and 150.

28. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:40, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163.

29. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 39, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 40, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100, 102, 103, 107, 141, 142 and 150.

30. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 16, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.

31. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14 and CDR3-H: SEQ ID NO: 16, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 107, 142 and 150.

32. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:50, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163.

33. An isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:49, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:50, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:107, 142 and 150.

34. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 22, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.

35. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14 and CDR3-H: SEQ ID NO: 22, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 107, 141, 142 and 150.

36. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:62, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119 and 123-163.

37. An isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:61, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:62, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:107, 141, 142 and 150.

38. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 26, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.

39. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14 and CDR3-H: SEQ ID NO: 26, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 107, 141, 142 and 150.

40. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 70, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119 and 123-163.

41. An isolated polypeptide or polypeptide complex according to claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:69, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:70, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:107, 141, 142 and 150.

42. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14, and CDR3-H: SEQ ID NO: 27, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163.

43. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a CDR set comprising CDR1-L: SEQ ID NO: 1, CDR2-L: SEQ ID NO: 2, CDR3-L: SEQ ID NO: 3, CDR1-H: SEQ ID NO: 13, CDR2-H: SEQ ID NO: 14 and CDR3-H: SEQ ID NO: 27, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, 158, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150, 158.

44. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:72, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:100-107, 109, 111-113, 116-117, 119 and 123-163.

45. The isolated polypeptide or polypeptide complex of claim 1, wherein the immunoglobulin light chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:71, and the immunoglobulin heavy chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:72, and wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs:107, 109, 116, 141, 142, 148, 149, 150 and 158, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs:107, 109, 116, 141, 142, 148, 149, 150 and 158.

46. ​​The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex further comprises a CD3 binding domain.

47. An isolated polypeptide or polypeptide complex according to claim 46, wherein the isolated polypeptide or polypeptide complex is according to the following formula P2-L2-B2-A1-L1-P1 (Formula Ia), wherein B2 comprises the CD3 binding domain, P2 comprises a peptide that binds to B2, and L2 comprises a linking portion that connects B2 to P2 and is a substrate for a tumor-specific protease.

48. The isolated polypeptide or polypeptide complex of claim 46, wherein the CD3 binding domain comprises an immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and an immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO:75; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO:76 (GTK); wherein CDR3-L comprises the amino acid sequence of SEQ ID NO:77; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:78; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:79; and Wherein CDR3-H comprises the amino acid sequence of SEQ ID NO:

80.

49. The isolated polypeptide or polypeptide complex of claim 46, wherein the CD3 binding domain comprises an immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and an immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO:75 or SEQ ID NO:259; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO:76 (GTK); wherein CDR3-L comprises the amino acid sequence of Z1-Z2-W-Z3-Z4-Z5-Z6-W-Z7-Z8; Where Z1 is V, G, P, L, I, M, S, T or A; Z2 is L, G, P, V, I, M, S, T, or A; Z3 is Y, F, W, V, L, I, G or A; Z4 is S, G, T, M, N, Q, H or A; Z5 is N, Q, S, T, D, E, H, K, R or A; Z6 is R, S, T, Q, D, E, H, K, N or A; Z7 is V, G, P, L, I, M, S, T, or A; and Z8 is F, Y, W, V, L, I, G, or A; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO:78 or SEQ ID NO:270; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO:79; and CDR3-H contains Z9-Z 10 -Z 11 -Z 12 -NZ 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -YZ 20 -AZ 21 The amino acid sequence of Where Z9 is V, G, P, L, I, M, S, T or A; Z 10 is R, S, T, Q, D, E, H, K, N or A; Z 11 is H, R, K, G, T, S, N, Q or A; Z 12 is G, P, V, L, I, M, S, T, or A; Z 13 is F, Y, W, V, L, I, G, or A; Z 14 is G, P, V, L, I, M, S, T, or A; Z 15 is N, Q, S, T, D, E, H, K, R or A; Z 16 is S, G, T, M, N, Q, H or A; Z 17 is Y, F, W, V, L, I, G or A; Z 18 is I, G, P, V, L, M, S, T, or A; Z 19 is S, G, T, M, N, Q, H or A; Z 20 is W, F, Y, V, L, I, G or A; and Z 21 It is Y, F, W, V, L, I, G or A.

50. The isolated polypeptide or polypeptide complex according to claim 49, wherein the CDR3-L of the CD3 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 77, 260-261, 263-266 and 268-269.

51. The isolated polypeptide or polypeptide complex of claim 49, wherein CDR3-H of the CD3 binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 80, 271-274, 276-282 and 284-285.

52. The isolated polypeptide or polypeptide complex of claim 46, wherein the CD3 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, the immunoglobulin light chain comprising complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and the immunoglobulin heavy chain comprising CDRs: CDR1-H, CDR2-H and CDR3-H, wherein the immunoglobulin light chain and the immunoglobulin heavy chain comprise a set of CDR sequences selected from the group consisting of: CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:260 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:261 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:262 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:263 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:264 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:265 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:266 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:267 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:268 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:269 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:80; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:271; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:272; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:273; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76(GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:274; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:275; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:276; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:277; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:278; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:279; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:280; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:281; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:282; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:283; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77, and CDR1-H: SEQ ID NO:78, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:284; CDR1-L: SEQ ID NO:75, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:77 and CDR1-H: SEQ ID NO: 78, CDR2-H: SEQ ID NO: 79, CDR3-H: SEQ ID NO: 285; and CDR1-L: SEQ ID NO:259, CDR2-L: SEQ ID NO:76 (GTK), CDR3-L: SEQ ID NO:260 and CDR1-H: SEQ ID NO:270, CDR2-H: SEQ ID NO:79, CDR3-H: SEQ ID NO:

286.

53. The isolated polypeptide or polypeptide complex of claim 46, wherein the CD3 binding domain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 99 and 303-329.

54. The isolated polypeptide or polypeptide complex of claim 46, wherein the CD3 binding domain comprises Fab, Fab', (Fab')2 or a single chain variable fragment (scFv).

55. The isolated polypeptide or polypeptide complex of claim 54, wherein the CD3 binding domain is the scFv.

56. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:87 and SEQ ID NO:

88.

57. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:89 and SEQ ID NO:

90.

58. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:91 and SEQ ID NO:

92.

59. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:93 and SEQ ID NO:

94.

60. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:95 and SEQ ID NO:

96.

61. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:97 and SEQ ID NO:

98.

62. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:330 and SEQ ID NO:

331.

63. An isolated polypeptide or polypeptide complex according to claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:332 and SEQ ID NO:

333.

64. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:334 and SEQ ID NO:

335.

65. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:336 and SEQ ID NO:

337.

66. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:338 and SEQ ID NO:

339.

67. An isolated polypeptide or polypeptide complex according to claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:340 and SEQ ID NO:

341.

68. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:342 and SEQ ID NO:

343.

69. An isolated polypeptide or polypeptide complex according to claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:344 and SEQ ID NO:

345.

70. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:346 and SEQ ID NO:

347.

71. An isolated polypeptide or polypeptide complex according to claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:348 and SEQ ID NO:

349.

72. The isolated polypeptide or polypeptide complex of claim 46, wherein the recombinant antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:350 and SEQ ID NO:

351.

73. The isolated polypeptide or polypeptide complex of claim 47, wherein P2 comprises the amino acid sequence of SEQ ID NO: 289 or SEQ ID NO:

292.

74. The isolated polypeptide or polypeptide complex according to claim 47, wherein P2 comprises an amino acid sequence according to any one of SEQ ID NOs: 287-302, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 287-302.

75. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 166 and SEQ ID NO:

167.

76. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 168 and SEQ ID NO:

169.

77. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 170 and SEQ ID NO:

171.

78. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 172 and SEQ ID NO:

173.

79. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 174 and SEQ ID NO:

175.

80. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 176 and SEQ ID NO:

177.

81. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 178 and SEQ ID NO:

179.

82. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 180 and SEQ ID NO:

181.

83. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 182 and SEQ ID NO:

183.

84. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 184 and SEQ ID NO:

185.

85. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 186 and SEQ ID NO:

187.

86. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 188 and SEQ ID NO:

189.

87. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 190 and SEQ ID NO:

191.

88. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 192 and SEQ ID NO:

193.

89. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 194 and SEQ ID NO:

195.

90. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 196 and SEQ ID NO:

197.

91. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 198 and SEQ ID NO:

199.

92. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 200 and SEQ ID NO:

201.

93. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:202 and SEQ ID NO:

203.

94. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:204 and SEQ ID NO:

205.

95. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:206 and SEQ ID NO:

207.

96. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:208 and SEQ ID NO:

209.

97. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:210 and SEQ ID NO:

211.

98. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:212 and SEQ ID NO:

213.

99. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:214 and SEQ ID NO:

215.

100. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:216 and SEQ ID NO:

217.

101. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:218 and SEQ ID NO:

219.

102. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:220 and SEQ ID NO:

221.

103. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:222 and SEQ ID NO:

223.

104. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:224 and SEQ ID NO:

225.

105. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:352 and SEQ ID NO:

353.

106. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:354 and SEQ ID NO:

355.

107. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:356 and SEQ ID NO:

357.

108. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:358 and SEQ ID NO:

359.

109. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:360 and SEQ ID NO:

361.

110. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:362 and SEQ ID NO:

363.

111. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:364 and SEQ ID NO:

365.

112. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:366 and SEQ ID NO:

367.

113. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:368 and SEQ ID NO:

369.

114. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:370 and SEQ ID NO:

371.

115. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:372 and SEQ ID NO:

373.

116. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:374 and SEQ ID NO:

375.

117. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:376 and SEQ ID NO:

377.

118. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:378 and SEQ ID NO:

379.

119. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:380 and SEQ ID NO:

381.

120. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:382 and SEQ ID NO:

383.

121. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:384 and SEQ ID NO:

385.

122. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:386 and SEQ ID NO:

387.

123. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:388 and SEQ ID NO:

389.

124. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:390 and SEQ ID NO:

391.

125. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:392 and SEQ ID NO:

393.

126. The isolated polypeptide or polypeptide complex of claim 1, wherein the isolated polypeptide or polypeptide complex comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:394 and SEQ ID NO:

395.

127. An isolated polypeptide or polypeptide complex according to formula I: A1-L1-P1 (Formula I) in: A1 comprises a recombinant antibody or an antigen-binding fragment thereof, wherein the recombinant antibody or the antigen-binding fragment thereof comprises a tumor-associated calcium signal transducer 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, wherein the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; wherein CDR3-L comprises the amino acid sequence of SEQ ID NO: 3; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and wherein CDR3-H comprises the amino acid sequence of SEQ ID NO:26; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163; and L1 comprises a linking moiety that connects A1 to P1 and is a substrate for tumor-specific proteases.

128. The isolated polypeptide or polypeptide complex according to claim 127, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 141, 142 and 150, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 107, 141, 142 and 150.

129. An isolated polypeptide or polypeptide complex according to formula I: A1-L1-P1 (Formula I) in: A1 comprises a recombinant antibody or an antigen-binding fragment thereof, wherein the recombinant antibody or the antigen-binding fragment thereof comprises a tumor-associated calcium signal transducer 2 (TROP2) binding domain, wherein the TROP2 binding domain comprises an immunoglobulin light chain and an immunoglobulin heavy chain, wherein the immunoglobulin light chain comprises complementarity determining regions (CDRs) CDR1-L, CDR2-L and CDR3-L, and the immunoglobulin heavy chain comprises CDRs: CDR1-H, CDR2-H and CDR3-H, wherein CDR1-L comprises the amino acid sequence of SEQ ID NO: 1; wherein CDR2-L comprises the amino acid sequence of SEQ ID NO: 2; wherein CDR3-L comprises the amino acid sequence of SEQ ID NO: 3; wherein CDR1-H comprises the amino acid sequence of SEQ ID NO: 13; wherein CDR2-H comprises the amino acid sequence of SEQ ID NO: 14; and wherein CDR3-H comprises the amino acid sequence of SEQ ID NO:27; P1 comprises a peptide that binds to A1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163, or an amino acid sequence having 1, 2, or 3 amino acid mutations, substitutions, or deletions relative to any one of SEQ ID NOs: 100-107, 109, 111-113, 116-117, 119, and 123-163; and L1 comprises a linking moiety that connects A1 to P1 and is a substrate for tumor-specific proteases.

130. An isolated polypeptide or polypeptide complex according to claim 129, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150 and 158, or an amino acid sequence having 1, 2 or 3 amino acid mutations, substitutions or deletions relative to any one of SEQ ID NOs: 107, 109, 116, 141, 142, 148, 149, 150 and 158.

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