CN116997573A - Antibodies targeting PSMA and CD3 and uses thereof - Google Patents

Antibodies targeting PSMA and CD3 and uses thereof Download PDF

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CN116997573A
CN116997573A CN202180084823.5A CN202180084823A CN116997573A CN 116997573 A CN116997573 A CN 116997573A CN 202180084823 A CN202180084823 A CN 202180084823A CN 116997573 A CN116997573 A CN 116997573A
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amino acid
seq
acid sequence
variable domain
fab
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大卫·坎贝尔
托马斯·R·迪莱蒙多
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Janux Therapeutics Inc
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Janux Therapeutics Inc
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Priority claimed from PCT/US2021/054948 external-priority patent/WO2022081822A1/en
Publication of CN116997573A publication Critical patent/CN116997573A/en
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Abstract

Provided herein are antibodies that selectively bind to PSMA and CD3, pharmaceutical compositions thereof, and methods of producing such antibodies.

Description

Antibodies targeting PSMA and CD3 and uses thereof
Cross reference
The present application claims the benefit of U.S. provisional application No. 63/092,226 filed on 10, 15, 2020 and U.S. provisional application No. 63/188,855 filed on 5, 2021, 14, each of which is incorporated herein by reference.
Sequence listing
The present application includes a sequence listing that is electronically submitted in ASCII format and hereby incorporated by reference in its entirety. The ASCII copy was created at 2021, 10 months and 12 days, named 52426-723_601_sl.txt, with a size of 43,741 bytes.
Disclosure of Invention
Disclosed herein are isolated polypeptide complexes according to the formula: a-L-B ((formula I), wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; B comprises an antigen binding fragment (Fab) or Fab 'bound to PSMA, wherein the Fab or Fab' comprises a Fab light chain polypeptide comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain, said Fab heavy chain variable domain comprising Complementarity Determining Regions (CDRs) HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise HC-CDR1: SEQ ID NO:17, HC-CDR2: SEQ ID NO:18 and HC-CDR3: SEQ ID NO:19; or HC-CDR1: SEQ ID NO:20, HC-CDR2: 18 and HC-CDR3: SEQ ID NO:21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR3, and L comprises a linker connecting A to B, in some embodiments, the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs) LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO:22, LC-CDR2: SEQ ID NO:23 and LC-CDR3: SEQ ID NO:24, or LC-CDR1: SEQ ID NO:25, LC-CDR2: SEQ ID NO:26 and LC-CDR3: SEQ ID NO:27, and wherein the CDRs are in the LC-CDR1, LC-CDR2 and LC-CDR3, at least one of the LC-CDR2 or LC-CDR3 comprises 0-2 amino acid modifications. In some embodiments, the scFv comprises a scFv light chain variable domain and a scFv heavy chain variable domain. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3. In some embodiments, the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 17 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence according to SEQ ID NO. 12 or 15. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the linker connects the C-terminus of a to the N-terminus of B. In some embodiments, the linker connects the N-terminus of a to the C-terminus of B. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the N-terminus of a to the C-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the N-terminus of a to the C-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the Fab light chain polypeptide to the scFv light chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the scFv light chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the N-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the N-terminus of the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain. In some embodiments, the linker is at least 5 amino acids in length. In some embodiments, the linker is no more than 30 amino acids in length. In some embodiments, the linker is at least 5 amino acids in length and no more than 30 amino acids in length. In some embodiments, the linker is 5 amino acids in length. In some embodiments, the linker is 15 amino acids in length. In some embodiments, the linker comprises SEQ ID NO:32 ((GGGGSGGGGSGGS) or SEQ ID NO:33 ((GGGGGGS). In some embodiments, a linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID NO:28 and is connected to the C-terminus of the Fab heavy chain variable domain, in some embodiments the amino acid sequence of the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID NO: 34. In some embodiments, a linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the amino acid sequence of at least 200, and is connected to the amino acid sequence of the Fab heavy chain variable domain of at least 200, and the amino acid sequence of the Fab heavy chain variable domain of at least 200, amino acid sequence of the Fab heavy chain variable domain of at least 28 comprises at least 100 consecutive amino acid residues of the amino acid sequence of the Fab polypeptide, and at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28, and the amino acid sequence of the Fab heavy chain polypeptide which is linked to the C-terminal end of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO. 34, and at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID NO. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 35. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 36. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID No. 38.
Disclosed herein are isolated polypeptide complexes according to the formula: a-L-D (formula II), wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D. In some embodiments, a Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain. In some embodiments, the scFv comprises a scFv light chain variable domain and a scFv heavy chain variable domain. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab light chain polypeptide. In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18, HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18, HC-CDR3: SEQ ID NO. 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3. In some embodiments, the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22; LC-CDR2: SEQ ID NO. 23; and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3. In some embodiments, the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 8 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence according to SEQ ID NO. 12 or 15. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the linker is at least 5 amino acids in length. In some embodiments, the linker is no more than 30 amino acids in length. In some embodiments, the linker is at least 5 amino acids in length and no more than 30 amino acids in length. In some embodiments, the linker is 5 amino acids in length. In some embodiments, the linker is 15 amino acids in length. In some embodiments, the linker comprises SEQ ID NO:32 ((GGGGSGGGGSGGS) or SEQ ID NO:33 ((GGGGGGS). In some embodiments, a linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID NO:28 and is connected to the C-terminus of the Fab heavy chain variable domain, in some embodiments the amino acid sequence of the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID NO: 34. In some embodiments, a linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the amino acid sequence of at least 200 of the amino acid sequence of the Fab heavy chain variable domain of the scFv heavy chain variable domain comprises at least 200, and is connected to the amino acid sequence of at least 200 consecutive amino acid residues of the Fab heavy chain variable domain of the scFv 28, and at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28, and the amino acid sequence of the Fab heavy chain polypeptide which is linked to the C-terminal end of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO. 34, and at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID NO. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:30 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 35. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 36. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID No. 38.
In certain embodiments, disclosed herein are pharmaceutical compositions comprising: an isolated polypeptide complex as described herein; and a pharmaceutically acceptable excipient.
In certain embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides of a polypeptide complex as described herein.
Drawings
The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:
FIGS. 1A-1C illustrate exemplary configurations of antibodies that selectively bind to PSMA and CD 3. FIG. 1A illustrates a Fab which binds to PSMA and an scFv which binds to CD3, wherein the N-terminus of the Fab heavy chain polypeptide is linked to the C-terminus of the scFv heavy chain variable domain by a linker. FIG. 1B illustrates a Fab that binds to PSMA and an scFv that binds to CD3, wherein the N-terminus of the Fab heavy chain polypeptide is linked to the C-terminus of the scFv light chain variable domain by a linker. FIG. 1C illustrates a Fab which binds to PSMA and an scFv which binds to CD3, wherein the N-terminus of the Fab light chain polypeptide is linked to the C-terminus of the scFv light chain variable domain by a linker.
FIG. 2 shows Ab-2 and Ab-3 binding to PSMA-biotin as measured by ELISA. Ab-3 has an EC50 of 0.16 nM. Ab-2 has an EC50 of 0.19 nM.
Figure 3 shows the titration data for PSMA binding of Ab-2.
FIG. 4 shows the titration data for PSMA binding of Ab-3.
FIG. 5 shows titration data for CD3 epsilon binding of Ab-2.
FIG. 6 shows titration data for CD3 epsilon binding of Ab-3.
FIG. 7 shows cytotoxicity of Ab-2 and Ab-3 as assessed in a cell viability assay in LNCaP tumor cells at 24 hours.
FIG. 8 shows cytotoxicity of Ab-2 and Ab-3 as assessed in a cell viability assay in LNCaP tumor cells at 48 hours.
FIG. 9 shows polypeptide complex mediated killing of 22Rv1 tumor cells in the presence of CD8+ T cells.
FIG. 10 shows polypeptide complex mediated killing of LNCaP tumor cells in the presence of CD8+ T cells.
Figure 11 shows the polypeptide pharmacokinetics in cynomolgus monkey bodies after a single IV bolus injection.
FIGS. 12A-12F show cytokine release in cynomolgus monkeys after a single IV bolus of TCE, IFN-gamma ((FIG. 12A), TNF-alpha (FIG. 12B), IL-6 ((FIG. 12C), IL-5 ((FIG. 12D), plasma IL-4 ((FIG. 12E), plasma IL-2 (FIG. 12F)).
Fig. 13 shows serum liver enzymes in cynomolgus monkeys after a single IV bolus of TCE.
FIG. 14 shows Ab-4 and Ab-5 binding to PSMA-biotin as measured by ELISA. Ab-4 has an EC50 of 2.8 nM. Ab-5 has an EC50 of 2.0 nM.
FIG. 15 shows the binding of Ab-4 and Ab-5 to CD 3-biotin as measured by ELISA. Ab-4 has an EC50 of 0.10 nM. Ab-5 has an EC50 of 0.17 nM.
Detailed Description
Multispecific antibodies combine the benefits derived from different binding specificities of two or more antibodies into a single composition. Multispecific antibodies for redirecting T cells to cancer have shown promise in both preclinical and clinical studies. This approach relies on the binding of one antigen-interacting moiety of an antibody to a tumor-associated antigen or marker, while a second antigen-interacting moiety can bind to an effector cell antigen on T cells, such as CD3, which then triggers cytotoxic activity.
One such tumor associated antigen is PSMA. Prostate Specific Membrane Antigen (PSMA), also known as glutamate carboxypeptidase II (GCPII), N-acetyl-L-aspartyl-L-glutamate peptidase I (NAALADase I), or nag peptidase, is an enzyme encoded by the FOLH1 (folate hydrolase 1) gene in humans. PSMA is a zinc metalloenzyme residing in a membrane. Most enzymes reside in the extracellular space. Human PSMA is highly expressed in the prostate gland, approximately 100-fold in most other tissues. In some prostate cancers, PSMA is the second largest upregulated gene product, increasing in levels 8-12 fold compared to non-cancerous prostate cells.
Disclosed herein are antibodies that selectively bind to PSMA and CD3, wherein the anti-PSMA domain is in the form of a Fab or Fab' antibody linked to a single chain variable fragment (scFv) that binds to CD 3. Bispecific antibody formats of Fab or Fab' linked to scFv offer efficacy and safety advantages over other bispecific antibody formats. In some embodiments, a Fab or Fab' comprises a Fab light chain polypeptide comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 21; and wherein the CDR comprises 0-2 amino acid modifications in at least one of the HC-CDR1, HC-CDR2 or HC-CDR 3. In some embodiments, the scFv that binds to CD3 is linked to the N-terminus of Fab or Fab' that binds to PSMA.
In some embodiments, the antibodies described herein are used in a method of treating cancer. In some embodiments, the cancer has PSMA-expressing cells. In some cases, the cancer is a solid tumor cancer. In some embodiments, the cancer is lung cancer, breast cancer (e.g., HER2+; ER/PR+; TNBC), cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer, or gastric cancer. In some embodiments, a polypeptide or polypeptide complex described herein is used in a method of treating prostate cancer. In some embodiments, the prostate cancer is metastatic castration-resistant prostate cancer (mCRPC). Prostate cancer is the second most common cancer among men worldwide, with more than 300 tens of thousands of men suffering from prostate cancer in the united states alone. Early diagnosis and effective therapy means that most prostate cancer patients have a prognosis with an average five-year survival of about 98%. However, it is estimated that 6% of prostate cancer patients develop metastatic disease, which is associated with five-year survival rates of about 30%. It is estimated that 33,000 people in the united states die in 2020 due to prostate cancer.
Certain definitions
The terminology used herein is for the purpose of describing particular instances only and is 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.
The term "antibody" is used in its broadest sense and encompasses fully assembled antibodies, antibody fragments that can bind to an antigen, such as Fab, F (ab') 2, fv, single chain antibodies (scFv), diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, and the like.
The term "complementarity determining region" or "CDR" is a segment of the variable region of an antibody that is structurally complementary to the epitope to which the antibody binds and is more variable than the remainder of the variable region. Thus, CDRs are sometimes referred to as hypervariable regions. The variable region comprises three CDRs. CDR peptides can be obtained by constructing genes encoding CDRs of an antibody of interest. Such genes are prepared, for example, by synthesizing variable regions from RNA of antibody-producing cells using polymerase chain reaction. See, for example, larrick et al Methods: A Companion to Methods in Enzymology 2:106 (1991); courtenay-Luck, "Genetic Manipulation of Monoclonal Antibodies", monoclonal Antibodies: production, engineering and Clinical Application, ritter et al (editorial) pages 166-179 (Cambridge university Press (Cambridge University Press) 1995), and Ward et al, "Genetic Manipulation and Expression of Antibodies", monoclonal Antibodies: principles and Applications, birch et al (editorial) pages 137-185 (Wiley-Lists, 1995).
The term "Fab" refers to a protein containing the constant domain of the light chain and the first constant domain of the heavy chain (CH 1). Fab fragments differ from Fab' fragments in that several residues are added at the carboxy terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab '-SH is herein the name of Fab', wherein the cysteine residues of the constant domain carry a free thiol group. Fab 'fragments are produced by reduction of the heavy chain disulfide bridges of F (ab') 2 fragments. Other chemical couplings of antibody fragments are also known.
A "single chain variable fragment (scFv)" is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of an antibody, linked to a short linker peptide of 10 to about 25 amino acids. The linker is typically rich in glycine for flexibility, as well as serine or threonine for solubility, and can link the N-terminus of VH with the C-terminus of VL, and vice versa. The protein retains the original antibody specificity, although the constant region is removed and a linker is introduced. scFv antibodies are described, for example, in Houston, j.s., methods in enzymol.203 (1991) 46-96. Furthermore, antibody fragments comprise single chain polypeptides having the characteristics of VH domains, i.e. capable of assembling with VL domains; or a single chain polypeptide featuring a VL domain, i.e., capable of assembling with a VH domain to a functional antigen binding site, thereby providing the antigen binding properties of a full length antibody.
As used herein, the term "percent (%) amino acid sequence identity" in relation to a sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular sequence after aligning the sequences and introducing gaps (gaps), if necessary, to obtain the maximum percent sequence identity, and does not take into account any conservative substitutions as part of the sequence identity. The alignment used to determine the percent amino acid sequence identity can be accomplished in a variety of ways within the skill of the art, for example, using publicly available computer software such as EMBOSS MATCHER, EMBOSS WATER, EMBOSS STRETCHER, EMBOSS NEEDLE, EMBOSS LALIGN, BLAST-2, ALIGN, or Megalign (DNASTAR) software. One skilled in the art can determine appropriate parameters for measuring the alignment, including any algorithms needed to achieve maximum alignment over the full length sequences being compared.
In the case of ALIGN-2 for amino acid sequence comparison, the amino acid sequence identity of a given amino acid sequence A relative to a given amino acid sequence B, to a given amino acid sequence B or to a given amino acid sequence B (which may alternatively be expressed as having or comprising a given amino acid sequence A) relative to a given amino acid sequence B, to a given amino acid sequence B or to a particular amino acid sequence identity% of a given amino acid sequence B is calculated as follows: a 100-fold score X/Y, wherein X is the number of amino acid residues recorded as the same match by sequence alignment program ALIGN-2 in the a and B alignments of the program, and wherein Y is the total number of amino acid residues in B. It is understood that in the case where 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 is not equal to the% amino acid sequence identity of B to a. All amino acid sequence identity% values used herein are obtained as described in the previous paragraph using the ALIGN-2 computer program, unless specifically indicated otherwise.
The terms "complementarity determining region" and "CDR," synonymous with "hypervariable region" or "HVR," are known in the art to refer to non-contiguous amino acid sequences within the variable region of an antibody that confer antigen specificity and/or binding affinity. Generally, 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 regions" and "FR" are known in the art to refer to the non-CDR portions of the variable regions of the heavy and light chains. In general, there are four FRs in each full length heavy chain variable region ((FR-H1, FR-H2, FR-H3, and FR-H4), and four FRs in each full length light chain variable region ((FR-L1, FR-L2, FR-L3, and FR-L4). The exact amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well known protocols, including Kabat et Al (1991), "Sequences of Proteins of Immunological Interest", 5 th edition, national institutes of health public health service center (Public Health Service, national Institutes of Health), bethesda, MD) in Maryland (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), "anti-body-antigen interactions: contact analysis and binding site topography", J.mol. Biol.262,732-745 "(" Contact "numbering scheme)," Lefranc MP et Al, "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains", dev Comp Immunol, 1 month 2003; 27 (1): 55-77 (("IMGT" numbering scheme), "Honyger A and Pluckthun A", "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool", J Mol Biol, 6 month 8 day 2001), "309 (3): 657-70" ("Aho" numbering scheme),; whitelegg NR and Rees AR), "WAM: an improved algorithm for modelling antibodies on the WEB", protein Eng.12, 2000; 13 819-24 ("AbM" numbering scheme). In certain embodiments, the CDRs of an antibody described herein can be defined by a method selected from Kabat, chothia, IMGT, aho, abM or a combination thereof.
The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. Numbering of both Kabat and Chothia protocols is based on the most common antibody region sequence length, with insertions regulated by insert letters such as "30a" and deletions occurring in some antibodies. These two schemes place certain insertions and deletions ("indels") at different positions, resulting in different numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme.
Disclosed herein are isolated polypeptide complexes according to the formula:
A-L-B
(formula I)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; and B comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B. Disclosed herein are isolated polypeptide complexes comprising the formula:
A-L-B
(formula I)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; and B comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain, the Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B. Disclosed herein are isolated polypeptide complexes comprising the formula:
A-L-B
(formula I)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; and B is an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L is a linker connecting a to B. Disclosed herein are isolated polypeptide complexes according to the formula:
A-L-B
(formula I)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; and B is an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L is a linker connecting a to B.
Disclosed herein are isolated polypeptide complexes according to the formula:
A-L-D
(formula II)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D. Disclosed herein are isolated polypeptide complexes comprising the formula:
A-L-D
(formula II)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D. Disclosed herein are isolated polypeptide complexes comprising the formula:
A-L-D
(formula II)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; d is an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L is a linker connecting the C-terminus of a to the N-terminus of D. Disclosed herein are isolated polypeptide complexes according to the formula:
A-L-D
(formula II)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; d is an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L is a linker connecting the C-terminus of a to the N-terminus of D.
Single chain variable fragment (scFv) binding to CD3
In some embodiments, the scFv that binds to CD3 comprises a scFv light chain variable domain and a scFv heavy chain variable domain. In some embodiments, the scFv heavy chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 1 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the scFv light chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 2 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).
In some embodiments, the scFv heavy chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 1 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and the scFv light chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 2 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).
In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3. In some embodiments, the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 4; HC-CDR2: SEQ ID NO. 2; HC-CDR3: 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 9; LC-CDR2: SEQ ID NO. 8; and LC-CDR3: SEQ ID NO. 10; and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1, HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1; HC-CDR2: SEQ ID NO. 2; HC-CDR3: SEQ ID NO. 3. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 4; HC-CDR2: SEQ ID NO. 2; HC-CDR3: SEQ ID NO. 5. In some embodiments, the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10. In some embodiments, the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 6; LC-CDR2: SEQ ID NO. 7; and LC-CDR3: SEQ ID NO. 8. In some embodiments, the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 9; LC-CDR2: SEQ ID NO. 7; and LC-CDR3: SEQ ID NO. 10.
In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1, HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5; and the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1; HC-CDR2: SEQ ID NO. 2; HC-CDR3: SEQ ID NO. 3; and the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 6; LC-CDR2: SEQ ID NO. 7; and LC-CDR3: SEQ ID NO. 8. In some embodiments, the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 4; HC-CDR2: SEQ ID NO. 2; HC-CDR3: SEQ ID NO. 5; and the scFv light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the scFv light chain variable domain comprise: LC-CDR1: SEQ ID NO. 9; LC-CDR2: SEQ ID NO. 7; and LC-CDR3: SEQ ID NO. 10.
TABLE 1 Complementarity Determining Regions (CDRs) of heavy chain variable domains of anti-CD 3 scFv (e.g., based on IMGT CDR numbering system).
TABLE 2 Complementarity Determining Regions (CDRs) of the light chain variable domains of anti-CD 3 scFv (e.g., based on the IMGT CDR numbering system).
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 70% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 85% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 91% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 92% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 93% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 94% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence according to SEQ ID NO. 12 or 15.
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 110 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 115 consecutive amino acid residues of SEQ ID NO. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 120 consecutive amino acid residues of SEQ ID NO. 12 or 15.
In some embodiments, the scFv heavy chain variable domain comprises the amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 60 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 70 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises the amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 80 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 90 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15.
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 60 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 70 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises the amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 80 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 90 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 90% sequence identity to at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15.
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 60 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 70 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises the amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 80 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 90 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 95% sequence identity to at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 99% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 99% sequence identity to at least 110 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 99% sequence identity to at least 115 consecutive amino acid residues of SEQ ID No. 12 or 15. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence of at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 99% sequence identity to at least 120 consecutive amino acid residues of SEQ ID No. 12 or 15.
In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 70% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 85% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 91% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 92% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 93% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 94% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 96% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence according to SEQ ID No. 11 or 14.
In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14.
In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises the amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises the amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14.
In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises the amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 90% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14.
In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 60 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 70 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises the amino acid sequence of at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 80 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 90 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 95% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14. In some embodiments, the scFv light chain variable domain comprises an amino acid sequence of at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 99% sequence identity to at least 105 consecutive amino acid residues of SEQ ID No. 11 or 14.
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 70% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 70% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 85% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 85% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 91% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 91% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 92% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 92% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 93% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 93% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 94% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 94% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 96% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14.
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 91% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 91% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 92% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 92% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 93% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 93% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 94% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 94% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 11. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID No. 12; and the scFv light chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 11.
In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 70% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 85% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 91% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 91% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 92% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 92% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 93% sequence identity to the amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 93% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 94% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 94% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 14. In some embodiments, the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID No. 15; and the scFv light chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 14.
In some embodiments, the scFv comprises an amino acid sequence having at least 70% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 85% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 90% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 91% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 92% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 93% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 94% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 95% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 96% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 97% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 98% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence having at least 99% sequence identity to an amino acid sequence according to SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence according to SEQ ID NO. 13 or 16.
In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16.
In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 80% sequence identity to at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16.
In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 90% sequence identity to at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16.
In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 95% sequence identity to at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16.
In some embodiments, the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 225 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 230 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 235 consecutive amino acid residues of SEQ ID NO. 13 or 16. In some embodiments, the scFv comprises an amino acid sequence of at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16, and has at least 99% sequence identity to at least 240 consecutive amino acid residues of SEQ ID NO. 13 or 16.
TABLE 3 anti-CD 3 scFv light chain variable Domain, heavy chain variable Domain sequence and full Length sequence
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Antigen binding fragments (Fab) or Fab 'bound to PSMA'
In some embodiments, an antigen binding fragment (Fab) or Fab' that binds to PSMA comprises a Fab light chain polypeptide chain and a Fab heavy chain polypeptide. In some embodiments, the Fab light chain polypeptide comprises a Fab light chain variable domain. In some embodiments, the Fab heavy chain polypeptide comprises a Fab heavy chain variable domain. In some embodiments, the Fab heavy chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 4 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the Fab light chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 5 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).
In some embodiments, the Fab heavy chain variable domain comprises or is substantially identical to at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 4 (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and the Fab light chain variable domain comprises at least one, two, or three Complementarity Determining Regions (CDRs) disclosed in table 5, or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3. In some embodiments, the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise: LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise: LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 21. In some embodiments, the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 21; and the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19, and the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise: LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24.
In some embodiments, the Fab heavy chain variable domain comprises Complementarity Determining Regions (CDRs): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 21; and the Fab light chain variable domain comprises Complementarity Determining Regions (CDRs): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise: LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27.
Table 4. Complementarity Determining Regions (CDRs) of the heavy chain variable domains of the anti-PSMA Fab (e.g., based on the IMGT CDR numbering system).
Table 5. Complementarity Determining Regions (CDRs) of the anti-PSMA Fab light chain variable domains (e.g., based on the IMGT CDR numbering system).
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 85% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 90% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 91% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 92% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 93% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 94% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 95% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 96% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 97% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 98% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 99% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence according to SEQ ID NO. 29 or 31.
In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 80% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31.
In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 90% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 95% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 29 or 31.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence of at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 215 consecutive amino acid residues of SEQ ID NO. 29 or 31. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31 and has at least 99% sequence identity to at least 220 consecutive amino acid residues of SEQ ID NO. 29 or 31.
In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 85% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 90% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 91% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 92% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 93% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 94% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 95% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 96% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 97% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 98% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence that has at least 99% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID NO. 28 or 30.
In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30.
In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 80% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30.
In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 90% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30.
In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 95% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30.
In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 150 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 175 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 190 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises an amino acid sequence of at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 205 consecutive amino acid residues of SEQ ID NO. 28 or 30. In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30 and has at least 99% sequence identity to at least 210 consecutive amino acid residues of SEQ ID NO. 28 or 30.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 70% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 80% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 85% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 90% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 90% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 91% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 91% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 92% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 92% sequence identity to the amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 93% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 93% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 94% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 94% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 95% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 95% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 96% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 96% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 97% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 97% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 98% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 98% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 99% sequence identity to an amino acid sequence according to SEQ ID NO. 29 or 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 99% sequence identity to an amino acid sequence according to SEQ ID NO. 28 or 30.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 70% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 70% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 80% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 85% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 85% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 90% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 91% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 91% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 92% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 92% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 93% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 93% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 94% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 94% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 95% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 96% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 96% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 97% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 97% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 98% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 98% sequence identity to the amino acid sequence according to SEQ ID NO. 28. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 99% sequence identity to the amino acid sequence according to SEQ ID NO. 29; and the Fab light chain polypeptide comprises an amino acid sequence with at least 99% sequence identity to the amino acid sequence according to SEQ ID NO. 28.
In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 70% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 70% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 80% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 80% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 85% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 85% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 90% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 91% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 91% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 92% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 92% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 93% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 93% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 94% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 94% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 95% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 96% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 96% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 97% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 97% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 98% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 98% sequence identity to the amino acid sequence according to SEQ ID NO. 30. In some embodiments, the Fab heavy chain polypeptide comprises an amino acid sequence that has at least 99% sequence identity to the amino acid sequence according to SEQ ID NO. 31; and the Fab light chain polypeptide comprises an amino acid sequence with at least 99% sequence identity to the amino acid sequence according to SEQ ID NO. 30.
TABLE 6 anti-PSMA Fab light chain polypeptide and Fab heavy chain polypeptide sequence
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Joint
In some embodiments, the linker is at least 5 amino acids in length. In some embodiments, the linker is no more than 30 amino acids in length. In some embodiments, the linker is at least 5 amino acids in length and no more than 30 amino acids in length. In some embodiments, the linker is 5 amino acids in length. In some embodiments, the linker is 6 amino acids in length. In some embodiments, the linker is 7 amino acids in length. In some embodiments, the linker is 8 amino acids in length. In some embodiments, the linker is 9 amino acids in length. In some embodiments, the linker is 10 amino acids in length. In some embodiments, the linker is 11 amino acids in length. In some embodiments, the linker is 12 amino acids in length. In some embodiments, the linker is 13 amino acids in length. In some embodiments, the linker is 14 amino acids in length. In some embodiments, the linker is 15 amino acids in length. In some embodiments, the linker is 16 amino acids in length. In some embodiments, the linker is 17 amino acids in length. In some embodiments, the linker is 18 amino acids in length. In some embodiments, the linker is 19 amino acids in length. In some embodiments, the linker is 20 amino acids in length. In some embodiments, the linker is 21 amino acids in length. In some embodiments, the linker is 22 amino acids in length. In some embodiments, the linker is 23 amino acids in length. In some embodiments, the linker is 24 amino acids in length. In some embodiments, the linker is 25 amino acids in length. In some embodiments, the linker is 26 amino acids in length. In some embodiments, the linker is 27 amino acids in length. In some embodiments, the linker is 28 amino acids in length. In some embodiments, the linker is 29 amino acids in length. In some embodiments, the linker is 30 amino acids in length. In some embodiments, the linker comprises the amino acid sequence of SEQ ID NO. 32 (GGGGSGGGGSGGGGS) or SEQ ID NO. 33 ((GGGGS).
In some embodiments, the linker connects the C-terminus of a to the N-terminus of B. In some embodiments, the linker connects the N-terminus of a to the C-terminus of B. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the N-terminus of a to the C-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the N-terminus of a to the C-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the Fab light chain polypeptide to the scFv light chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the scFv light chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the N-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the N-terminus of the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv heavy chain variable domain. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain.
In some embodiments, the linker connects the C-terminus of a to the N-terminus of D. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab light chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab heavy chain polypeptide. In some embodiments, the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab light chain polypeptide.
TABLE 7 linker sequences
Antibodies that bind to PSMA and CD3
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34 and has at least 80% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 28 and has at least 80% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 34 and has at least 80% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34 and has at least 90% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34 and has at least 90% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 28 and has at least 90% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 34 and has at least 90% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34 and has at least 95% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34 and has at least 95% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 28 and has at least 95% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 34 and has at least 95% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34 and has at least 99% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34 and has at least 99% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 34. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 28 and has at least 99% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 34 and has at least 99% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 34.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID NO: 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 35 and has at least 90% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 90% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 90% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 35 and has at least 90% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 35 and has at least 95% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 95% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 95% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 35 and has at least 95% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 35 and has at least 99% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 99% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 99% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 35 and has at least 99% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 35.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 35.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 36.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID NO: 36.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 460 consecutive amino acid residues of SEQ ID No. 36.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 36 and has at least 90% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 90% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 90% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 36 and has at least 90% sequence identity to at least 460 consecutive amino acid residues of SEQ ID No. 36.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 36 and has at least 95% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 95% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 95% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 36 and has at least 95% sequence identity to at least 460 consecutive amino acid residues of SEQ ID No. 36.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 36 and has at least 99% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 99% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 99% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 36 and has at least 99% sequence identity to said at least 460 consecutive amino acid residues.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 36.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37 and has at least 90% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 90% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 90% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 37 and has at least 90% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37 and has at least 95% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 95% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 95% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 37 and has at least 95% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and has at least 99% sequence identity with at least 100 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37 and has at least 99% sequence identity with at least 400 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 99% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37. In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence of at least 210 consecutive amino acid residues of SEQ ID No. 30 and has at least 99% sequence identity to at least 210 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 470 consecutive amino acid residues of SEQ ID No. 37 and has at least 99% sequence identity to at least 470 consecutive amino acid residues of SEQ ID No. 37.
In some embodiments, the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID No. 37.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 96% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 97% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 98% sequence identity to the amino acid sequence according to SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 99% sequence identity to the amino acid sequence according to SEQ ID No. 38.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID NO: 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID NO: 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID NO:31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID NO: 38.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 460 consecutive amino acid residues of SEQ ID No. 38.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 90% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38 and has at least 90% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 90% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 90% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 90% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 38 and has at least 90% sequence identity to at least 460 consecutive amino acid residues of SEQ ID No. 38.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 95% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38 and has at least 95% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 95% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 95% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 95% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 38 and has at least 95% sequence identity to at least 460 consecutive amino acid residues of SEQ ID No. 38.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and has at least 99% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38 and has at least 99% sequence identity to at least 400 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 99% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 99% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38. In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence of at least 220 consecutive amino acid residues of SEQ ID No. 31 and has at least 99% sequence identity to at least 220 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 460 consecutive amino acid residues of SEQ ID No. 38 and has at least 99% sequence identity to said at least 460 consecutive amino acid residues.
In some embodiments, the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID No. 38.
TABLE 8 antibody sequences binding to PSMA and CD3
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Polynucleotides encoding polypeptides or polypeptide complexes
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes as disclosed herein. In some embodiments, described herein are isolated recombinant nucleic acid molecules encoding polypeptides comprising antibodies that selectively bind to CD3 and PSMA.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides of a polypeptide complex according to the formula:
A-L-B
(formula I)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; and B comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B. In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides comprising a polypeptide complex of the formula:
A-L-B
(formula I)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; and B comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B. In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides comprising a polypeptide complex of the formula:
A-L-B
(formula I)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; and B is an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L is a linker connecting a to B. In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides of a polypeptide complex according to the formula:
A-L-B
(formula I)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; and B is an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L is a linker connecting a to B.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides of a polypeptide complex according to the formula:
A-L-D
(formula II)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D. In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides comprising a polypeptide complex of the formula:
A-L-D
(formula II)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D. In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides comprising a polypeptide complex of the formula:
A-L-D
(formula II)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; d is an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L is a linker connecting the C-terminus of a to the N-terminus of D. In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding polypeptides of a polypeptide complex according to the formula:
A-L-D
(formula II)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; d is an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L is a linker connecting the C-terminus of a to the N-terminus of D.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 28.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 29.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 30.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 31.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 34.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 35.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 36.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 37.
In some embodiments, disclosed herein are isolated recombinant nucleic acid molecules encoding isolated polypeptides comprising amino acids according to SEQ ID NO. 38.
Pharmaceutical composition
In some embodiments, disclosed herein are pharmaceutical compositions comprising: (a) An isolated polypeptide or polypeptide complex as disclosed herein; and (b) a pharmaceutically acceptable excipient.
In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex according to formula I:
A-L-B
(formula I)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; and B comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex comprising formula I:
A-L-B
(formula I)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; and B comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex comprising formula I:
A-L-B
(formula I)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; and B is an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L is a linker connecting a to B; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex according to formula I:
A-L-B
(formula I)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; and B is an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L is a linker connecting a to B; and (b) a pharmaceutically acceptable excipient.
In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex according to formula I:
A-L-D
(formula II)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex comprising formula I:
A-L-D
(formula II)
Wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L comprises a linker connecting the C-terminus of a to the N-terminus of D; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex comprising formula I:
A-L-D
(formula II)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; d is an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L is a linker connecting the C-terminus of a to the N-terminus of D; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises: (a) an isolated polypeptide complex according to formula I:
A-L-D
(formula II)
Wherein a is a single chain variable fragment (scFv) that binds to CD 3; d is an antigen binding fragment (Fab) or Fab' that binds to PSMA; and L is a linker connecting the C-terminus of a to the N-terminus of D; and (b) a pharmaceutically acceptable excipient.
In some embodiments, the polypeptide or polypeptide complex further comprises a detectable label, therapeutic agent, or pharmacokinetic modifying moiety. In some embodiments, the detectable label comprises a fluorescent label, a radiolabel, an enzyme, a nucleic acid probe, or a contrast agent.
For administration to a subject, a polypeptide or polypeptide complex as disclosed herein may be provided in the form of a pharmaceutical composition together with one or more pharmaceutically acceptable carriers or excipients. The term "pharmaceutically acceptable carrier" includes, but is not limited to, any carrier that does not interfere with the efficacy of the biological activity of the ingredient and that is non-toxic to the patient to whom it is administered. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions (e.g., oil/water emulsions), various types of wetting agents, sterile solutions, and the like. Such carriers can be formulated by conventional methods and can be administered to a subject in appropriate dosages. Preferably, the composition is sterile. These compositions may also contain adjuvants such as preserving, emulsifying and dispersing agents. By including various antibacterial and antifungal agents, prevention of the action of microorganisms can be ensured.
The pharmaceutical composition may be in any suitable form (depending on the desired method of administration). It may be provided in unit dosage form, may be provided in a sealed container, and may be provided as part of a kit. Such a kit may include instructions for use. It may comprise a plurality of said unit dosage forms.
The pharmaceutical composition may be suitable for administration by any suitable route, including parenteral (e.g., subcutaneous, intramuscular, or intravenous) route. Such compositions may be prepared by any method known in the pharmaceutical arts, for example by mixing the active ingredient with a carrier or excipient under sterile conditions.
The dosage of the substances of the present disclosure may vary between wide limits, depending on the disease or disorder to be treated, the age and condition of the individual to be treated, etc., and the physician will ultimately determine the appropriate dosage to be used.
Therapeutic method
In some embodiments, is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a polypeptide or polypeptide complex as described herein. In some embodiments, the cancer has PSMA-expressing cells. In some cases, the cancer is a solid tumor cancer. In some embodiments, the cancer is lung cancer, breast cancer (e.g., HER2+; ER/PR+; TNBC), cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer, or gastric cancer.
In some embodiments, is a method of treating prostate cancer in a subject in need thereof, the method comprising administering to the subject a polypeptide or polypeptide complex as described herein. In some embodiments, is a method of treating metastatic castration-resistant prostate cancer (mCRPC) in a subject in need thereof, the method comprising administering to the subject a polypeptide or polypeptide complex as described herein.
Production of antibodies that bind to PSMA and CD3
In some embodiments, the polypeptides described herein (e.g., antibodies and binding fragments thereof) are produced using any method known in the art that can be used to synthesize polypeptides (e.g., antibodies), particularly by chemical synthesis or by recombinant expression, and preferably by recombinant expression techniques.
In some cases, the antibodies or binding fragments thereof are expressed recombinantly, and the nucleic acids encoding the antibodies or binding fragments thereof are assembled from chemically synthesized oligonucleotides (e.g., as described in Kutmeier et al, 1994,BioTechniques 17:242), which involves synthesizing overlapping oligonucleotides containing portions of the sequences encoding the antibodies, annealing and ligating the oligonucleotides, and then amplifying the ligated oligonucleotides by PCR.
Alternatively, the nucleic acid molecules encoding the antibodies are optionally generated from a suitable source (e.g., an antibody cDNA library, or a cDNA library generated from any tissue or cell expressing immunoglobulins) by PCR amplification using synthetic primers that hybridize to the 3 'and 5' ends of the sequences or by cloning using oligonucleotide probes specific for the particular gene sequences.
In some cases, the antibodies or binding thereof are optionally produced by immunizing an animal (e.g., a mouse) to produce polyclonal antibodies, or more preferably by producing monoclonal antibodies, e.g., as described by Kohler and Milstein (1975,Nature 256:495-497), or as described by Kozbor et al (1983,Immunology Today 4:72) or Cole et al (1985in Monoclonal Antibodies and Cancer Therapy,Alan R.Liss,Inc, pages 77-96). Alternatively, clones encoding at least the Fab portion of the antibody are optionally obtained by screening Fab expression libraries (e.g. as described in hule et al 1989,Science 246:1275-1281) against clones binding Fab fragments of the specific antigen or by screening antibody libraries (see, e.g. Clackson et al 1991,Nature 352:624;Hane et al 1997Proc.Natl.Acad.Sci.USA 94:4937).
In some embodiments, techniques developed for the production of "chimeric antibodies" by splicing together genes from mouse antibody molecules with appropriate antigen specificity and genes from human antibody molecules with appropriate biological activity are used (Morrison et al, 1984, proc. Natl. Acad. Sci.81:851-855; neuberger et al, 1984,Nature 312:604-608; takeda et al, 1985,Nature 314:452-454). Chimeric antibodies are molecules in which different portions are derived from different animal species, such as those having variable regions from murine monoclonal antibodies and human immunoglobulin constant regions.
In some embodiments, the techniques described for producing single chain antibodies (U.S. Pat. No. 4,694,778; bird,1988,Science 242:423-42; huston et al, 1988,Proc.Natl.Acad.Sci.USA 85:5879-5883; and Ward et al, 1989,Nature 334:544-54) are suitable for producing single chain antibodies. Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge to yield a single chain polypeptide. Techniques for assembling functional Fv fragments in E.coli are also optionally used (Skerra et al 1988,Science 242:1038-1041).
In some embodiments, the expression vector comprising the nucleotide sequence of the antibody or the nucleotide sequence of the antibody is transferred to a host cell by conventional techniques (e.g., electroporation, lipofection, and calcium phosphate precipitation), and the transfected cells are then cultured by conventional techniques to produce the antibody. In particular embodiments, expression of the antibody is regulated by a constitutive, inducible or tissue specific promoter.
In some embodiments, a variety of host expression vector systems are utilized to express the antibodies or binding fragments thereof described herein. Such host expression systems represent vehicles for the coding sequences of antibodies that are produced and subsequently purified, but also represent cells that express the antibodies or binding fragments thereof in situ when transformed or transfected with the appropriate nucleotide coding sequences. These include, but are not limited to, microorganisms such as bacteria (e.g., E.coli and B.subtilis) transformed with recombinant phage DNA, plasmid DNA, or cosmid DNA expression vectors containing the coding sequences for the antibodies or binding fragments thereof; yeast (e.g., pichia pastoris (Saccharomyces Pichia)) transformed with a recombinant yeast expression vector comprising a coding sequence for an antibody or binding fragment thereof; insect cell systems infected with recombinant viral expression vectors (e.g., baculovirus) containing sequences encoding antibodies or binding fragments thereof; plant cell systems infected with recombinant viral expression vectors (e.g., cauliflower mosaic virus (CaMV) and Tobacco Mosaic Virus (TMV)) or transformed with recombinant plasmid expression vectors (e.g., ti plasmid) containing the coding sequences of antibodies or binding fragments thereof; or mammalian cell systems (e.g., COS, CHO, BH, 293T, 3T3 cells) comprising recombinant expression constructs containing promoters from the genome of mammalian cells (e.g., metallothionein promoters) or promoters from mammalian viruses (e.g., adenovirus late promoter; vaccinia virus 7.5K promoter).
For long-term high-yield production of recombinant proteins, stable expression is preferred. In some cases, cell lines that stably express the antibody are optionally engineered. Instead of using an expression vector containing a viral origin of replication, the host cell is transformed with DNA controlled by suitable expression control elements (e.g., promoters, enhancers, sequences, transcription terminators, polyadenylation sites, etc.), and selectable markers. After introduction of the exogenous DNA, the engineered cells were allowed to grow in the enrichment medium for 1-2 days and then transferred to selective media. The selectable markers in the recombinant plasmids confer resistance to selection and allow the cells to stably integrate the plasmids into their chromosomes and grow to form foci, which in turn are cloned and expanded into cell lines. The method can be advantageously used to engineer cell lines expressing antibodies or binding fragments thereof.
In some cases, a number of selection systems have been used, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al, 1977, cell 11:223), hypoxanthine-guanine phosphoribosyl transferase (Szybalska & Szybalski,192,Proc.Natl.Acad.Sci.USA 48:202) and adenine phosphoribosyl transferase (Lowy et al, 1980, cell 22:817) genes for tk cells, hgprt cells or aprt cells, respectively. Furthermore, antimetabolite resistance is used as a basis for selection of the following genes: dhfr, which confers resistance to methotrexate (Wigler et al, 1980,Proc.Natl.Acad.Sci.USA 77:357;O'Hare et al, 1981,Proc.Natl.Acad.Sci.USA 78:1527); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg,1981,Proc.Natl.Acad.Sci.USA 78:2072); neo, which confers resistance to aminoglycoside G-418 (Clinical Pharmacy 12:488-505; wu and Wu,1991,Biotherapy 3:87-95; tolstoshaev, 1993, ann. Rev. Pharmacol. Toxicol.32:573-596; mulligan,1993, science260:926-932; and Morgan and Anderson,1993, ann. Rev. Biochem.62:191-217; 5 month 1993, TIB TECH 11 (5): 155-215) and hygro, which confers resistance to hygromycin (Santerre et al, 1984, gene 30:147). Methods well known in the art of recombinant DNA technology that can be used are described in Ausubel et al (editorial, 1993,Current Protocols in Molecular Biology, N.Y. John Willi parent (John Wiley & Sons, N.Y.), kriegler,1990,Gene Transfer and Expression,A Laboratory Manual, N.Y. Stoketon Press, N.Y.), and in chapters 12 and 13, dracopoli et al (editorial), 1994,Current Protocols in Human Genetics, N.Y. Willi parent, colberre-Garapin et al, 1981, J.mol. Biol. 150:1).
In some cases, the expression level of the antibody is increased by vector amplification (for reviews, see Bebbington and Hentschel, the use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, vol.3 (Academic Press, new York, 1987). When the markers in the vector system expressing the antibody are amplifiable, an increase in the level of inhibitor present in the culture of the host cell will increase the copy number of the marker gene. Since the amplified region correlates with the nucleotide sequence of the antibody, the yield of antibody will also increase (Crouse et al 1983,Mol.Cell Biol.3:257).
In some cases, any method known in the art for purifying antibodies is used, for example, by chromatography (e.g., ion exchange, affinity, in particular by affinity for specific antigens after protein a, and size column chromatography), centrifugation, differential lysis, or by any other standard technique for purifying proteins.
Expression vector
In some embodiments, the vector comprises any suitable vector derived from eukaryotic or prokaryotic sources. In some cases, the vector is obtained from a bacterial (e.g., E.coli), insect, yeast (e.g., pichia pastoris), algae, or mammalian source. Exemplary bacterial vectors include pACYC177, pASK75, pBAD vector series, pBADM vector series, pET vector series, pETM vector series, pGEX vector series, pHAT2, pMal-C2, pMal-p2, pQE vector series, pRSET A, pRSET B, pRSET C, pTrcHis2 series, pZA31-Luc, pZE21-MCS-1, pFLAG ATS, pFLAG CTS, pFLAG MAC, pFLAG Shift-12C, pTAC-MAT-1, pFLAG CTC or pTAC-MAT-2.
Exemplary insect vectors include pFastBac1, pFastBac DUAL, pFastBac ET, pFastBac HTa, pFastBac HTb, pFastBac HTc, pFastBac M30a, pFastBac M30b, pFastBac, M c, pVL1392, pVL1393M 10, pVL1393M11, pVL1393M 12, FLAG vectors such as pPolh-FLAG1 or pPolh-MAT2, or MAT vectors such as pPolh-MAT1 or pPolh-MAT2.
In some cases, the yeast carrier includespDEST TM 14 vector,>pDEST TM 15 vector, (-) ->pDEST TM 17 vector, (-) ->pDEST TM 24 vector,>pYES-DEST52 vector, pBAD-DEST 49->The vector of interest, pAO815 Pichia pastoris vector, pFLD1 Pichia pastoris vector, pGAPZA, B and C Pichia pastoris vector, pPICC 3.5K Pichia pastoris vector, pPIC 6A, B and C Pichia pastoris vector, pPIC9K Pichia pastoris vector, pTEF1/Zeo, pYES2 yeast vector, pYES2/CT yeast vector, pYES2/NT A, B and C yeast vector or pYES3/CT yeast vector.
Exemplary algal vectors include pChlamy-4 vectors or MCS vectors.
Examples of mammalian vectors include transient expression vectors or stable expression vectors. Mammalian transient expression vectors may include pRK5, p3xFLAG-CMV 8, pFAG-Myc-CMV 19, pFAG-Myc-CMV 23, pFAG-CMV 2, pFAG-CMV 6a, b, c, pFLAG-CMV 5.1, pFAG-CMV 5a, b, c, p3xFLAG-CMV 7.1, pFAG-CMV 20, p3xFLAG-Myc-CMV 24, pCMV-FLAG-MAT1, pCMV-FLAG-MAT2, pBICEP-CMV 3 or pBICEP-CMV 4. The mammalian stable expression vector may include pFLAG-CMV 3, p3xFLAG-CMV 9, p3xFLAG-CMV 13, pFLAG-Myc-CMV 21, p3xFLAG-Myc-CMV 25, pFLAG-CMV 4, p3xFLAG-CMV 10, p3xFLAG-CMV 14, pFLAG-Myc-CMV 22, p3xFLAG-Myc-CMV 26, pBICEP-CMV 1, or pBICEP-CMV 2.
In some cases, the cell-free system is a mixture of cytoplasmic and/or nuclear fractions from cells and is used for in vitro nucleic acid synthesis. In some cases, the cell-free system utilizes a prokaryotic or eukaryotic cell component. Sometimes, nucleic acid synthesis is obtained in cell-free systems based on, for example, drosophila cells, xenopus eggs or HeLa cells. Exemplary cell-free systems include, but are not limited to, the E.coli S30 extraction system, the E.coli T7S 30 system, orHost cells
In some embodiments, the host cell comprises any suitable cell, such as a naturally derived cell or a genetically modified cell. In some cases, the host cell is a production host cell. In some cases, the host cell is a eukaryotic cell. In other cases, the host cell is a prokaryotic cell. In some cases, eukaryotic cells include fungi (e.g., yeast cells), animal cells, or plant cells. In some cases, the prokaryotic cell is a bacterial cell. Examples of bacterial cells include gram positive bacteria or gram negative bacteria. Sometimes gram-negative bacteria are anaerobic, rod-shaped, or both.
In some cases, the gram-positive bacteria include actinomycota, firmicutes (Firmicutes), or tenebrio (Tenericutes). In some cases, gram-negative bacteria include aquaponics (aquifiae), anococcus-Thermus (Deinococcus-Thermus), cellomycota-viridans/bacteroides (fibrirobacters-chlorois/bacterioides) (FCB group), fusobacterium (fusobacterium), blastomyces (gemmatides), nitrifying spiras (nitrospiras), floating mold-warts/chlamydia (PVC group), proteus (protebacillus), spira (spiras) or cross-bacteria (bacteria) other bacteria may be Acidobacteria (aciobacter), viridae (chrysogens), cyanobacterium (c), bacteriosis (haemobacteria), coliform (haemobacteria), or coliform (haemobacteria), or coliform (haemobacteria).
Exemplary prokaryotic host cells include, but are not limited to BL21, mach1 TM 、DH10B TM 、TOP10、DH5α、DH10Bac TM 、OmniMax TM 、MegaX TM 、DH12S TM 、INV110、TOP10F’、INVαF、TOP10/P3、ccdB Survival、PIR1、PIR2、Stbl2 TM 、Stbl3 TM Or Stbl4 TM 。
In some cases, the animal cells include cells from vertebrates or invertebrates. In some cases, the animal cells include cells from marine invertebrates, fish, insects, amphibians, reptiles, or mammals. In some cases, the fungal cells include yeast cells, such as brewer's yeast, baker's yeast, or wine yeast.
Fungi include ascomycetes such as yeasts, molds, filamentous fungi, basidiomycetes or zygomycetes. In some cases, the yeast includes ascomycota or basidiomycota. In some cases, ascomycota includes phylum saccharomyces (true yeast, e.g., wine yeast (baker's yeast)) or exomycotina (e.g., schizosaccharomyces (schizosaccharomyces)), in some cases, basidiomycota includes phylum agaricus (e.g., tremella (tremellomyces)) or phylum puccinia (e.g., microsphere melanomyces).
Exemplary yeasts or filamentous fungi include, for example, the following genera: saccharomyces, schizosaccharomyces, candida, pichia, hansenula, kluyveromyces, zygosaccharomyces, yarrowia, trichosporon, rhodosporidium, aspergillus, fusarium, or Trichoderma. Exemplary yeasts or filamentous fungi include, for example, the following species: wine yeast, schizosaccharomyces pombe, candida utilis, candida boidinii (Candida boidinii), candida albicans, candida tropicalis, candida stellate ((Candida stellatoidea), candida glabra ((Candida glabra), candida krusei ((Candida krusei), candida parapsilosis, candida mongolica (Candida guilliermondii), candida vista (Candida viswanathii), candida staphylovora (Candida lusitaniae), rhodotorula mucilaginosa (Rhodotorula mucilaginosa), pichia methanolica (Pichia metanolica), pichia angusta (Pichia angusta), pichia pastoris, abnormal yeasts, hansenula polymorpha, kluyveromyces lactis, zygosaccharomyces rouxii (Zygosaccharomyces rouxii), yarrowia lipolytica, candida pullulans (Trichosporon pullulans), rhodotorula-niger, aspergillus nidulans, aspergillus awamori (Aspergillus awamori), candida oryzae (Aspergillus oryzae), trichoderma reesei, yarrowia lipolytica (Brettanomyces bruxellensis), candida rugosa (3765), candida rugosa (Cryptococcus gattii), candida nodosa (Cryptococcus gattii), candida nodorum (Cryptococcus gattii), candida albicans (Cryptococcus gattii).
Exemplary yeast host cells include, but are not limited to, pichia pastoris strains such as GS115, KM71H, SMD1168, SMD1168H and X-33; and wine yeast strains such as INVSc1.
In some cases, the additional animal cells include cells obtained from molluscs, arthropods, annelids, or sponges. In some cases, the additional animal cells are mammalian cells, such as cells from primates, apes, equines, bovine, porcine, canine, feline, or rodent. In some cases, the rodent comprises a mouse, rat, hamster, gerbil, hamster, castanea mollissima, gerbil, or guinea pig.
Exemplary mammalian host cells include, but are not limited to, 293A cell lines, 293FT cell lines, 293F cells, 293H cells, CHO DG44 cells, CHO-S cells, CHO-K1 cells, FUT8 KO CHOK1, expi293F TM Cells, flp-In TM T-REx TM 293 cell line, flp-In TM 293 cell line, flp-In TM 3T3 cell line, flp-In TM BHK cell line, flp-In TM CHO cell line, flp-In TM CV-1 cell line, flp-In TM Jurkat cell line, freeStyle TM 293-F cells, freeStyle TM CHO-S cells, gritite TM 293 MSR cell line, GS-CHO cell line, hepaRG TM Cells, T-REx TM Jurkat cell line, per.C6 cell, T-REx TM -293 cell line, T-REx TM CHO cell line and T-REx TM HeLa cell line.
In some cases, the mammalian host cell is a stable cell line, or a cell line that has incorporated the genetic material of interest into its own genome and has the ability to express the product of that genetic material after many generations of cell division. In some cases, the mammalian host cell is a transient cell line, or a cell line that does not incorporate the genetic material of interest into its own genome and does not have the ability to express the product of the genetic material after many passages of cell division.
Exemplary insect host cells include, but are not limited to, drosophila S2 cells, sf9 cells, sf21 cells, high Five cells TM Cells and methods of useAnd (3) cells.
In some cases, the plant cells include cells from algae. Exemplary insect cell lines include, but are not limited to, strains from chlamydomonas reinhardtii (Chlamydomonas reinhardtii) 137c or synechococcus elongatus (Synechococcus elongatus) PPC 7942.
Article of manufacture
In another aspect of the invention, an article of manufacture is provided that contains materials for the treatment, prevention and/or diagnosis of the above-described conditions. The article includes a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, and the like. The container may be formed from a variety of materials, such as glass or plastic. The container contains a composition that is effective, either by itself or in combination with another composition, for treating, preventing and/or diagnosing a condition; and may have a sterile access port (e.g., the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a bispecific antibody comprising a first antigen-binding site that specifically binds to CD3 and a second antigen-binding site that specifically binds to PSMA as defined above.
The label or package insert indicates that the composition is to be used to treat the selected condition. Furthermore, the article of manufacture may comprise (a) a first container having a composition contained therein, wherein the composition comprises a bispecific antibody of the present invention; and (b) a second container having a composition contained therein, wherein the composition comprises an additional cytotoxic agent or other therapeutic agent. The article of manufacture in this embodiment of the invention may further comprise a package insert indicating that the composition may be used to treat a particular condition.
Alternatively or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, ringer's solution, and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles and syringes.
Description of the embodiments
Embodiment 1 includes an isolated polypeptide complex according to the formula: a-L-B (formula I), wherein a comprises a single chain variable fragment (scFv) that binds to CD 3; b comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein Fab or Fab' comprises a Fab light chain polypeptide comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3; and L comprises a linker connecting a to B.
Embodiment 2 includes an isolated polypeptide complex according to embodiment 1, wherein the Fab light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
Embodiment 3 includes an isolated polypeptide complex according to any of embodiments 1-2, wherein the scFv comprises a scFv light chain variable domain and a scFv heavy chain variable domain.
Embodiment 4 includes an isolated polypeptide complex according to any of embodiments 1-3, wherein the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3.
Embodiment 5 includes an isolated polypeptide complex according to any of embodiments 1-4, wherein the scFv light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
Embodiment 6 includes an isolated polypeptide complex according to any of embodiments 1-5, wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 29 or 31.
Embodiment 7 includes an isolated polypeptide complex according to any of embodiments 1-6, wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO 29 or 31.
Embodiment 8 includes an isolated polypeptide complex according to any of embodiments 1-7, wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO 29 or 31.
Embodiment 9 includes the isolated polypeptide complex of any of embodiments 1-8, wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 29 or 31, and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 29 or 31.
Embodiment 10 includes an isolated polypeptide complex according to any of embodiments 1-9, wherein the Fab heavy chain polypeptide comprises an amino acid sequence according to SEQ ID No. 29 or 31.
Embodiment 11 includes the isolated polypeptide complex of any of embodiments 1-10, wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 28 or 30.
Embodiment 12 includes an isolated polypeptide complex according to any of embodiments 1-11, wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO 28 or 30.
Embodiment 13 includes an isolated polypeptide complex according to any of embodiments 1-12, wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID NO 28 or 30.
Embodiment 14 includes an isolated polypeptide complex according to any of embodiments 1-13, wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 or 30, and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28 or 30.
Embodiment 15 includes an isolated polypeptide complex according to any of embodiments 1-14, wherein the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID No. 28 or 30.
Embodiment 16 includes an isolated polypeptide complex according to any of embodiments 1-15, wherein the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15.
Embodiment 17 includes an isolated polypeptide complex according to any of embodiments 1-16, wherein the scFv heavy chain variable domain comprises the amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15.
Embodiment 18 includes an isolated polypeptide complex according to any of embodiments 1-17, wherein the scFv heavy chain variable domain comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15.
Embodiment 19 includes an isolated polypeptide complex according to any of embodiments 1-18, wherein the scFv heavy chain variable domain comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15.
Embodiment 20 includes an isolated polypeptide complex according to any of embodiments 1-19, wherein the scFv heavy chain variable domain comprises an amino acid sequence according to SEQ ID No. 12 or 15.
Embodiment 21 includes an isolated polypeptide complex according to any of embodiments 1-20, wherein the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14.
Embodiment 22 includes an isolated polypeptide complex according to any of embodiments 1-20, wherein the scFv light chain variable domain comprises the amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14.
Embodiment 23 includes an isolated polypeptide complex according to embodiment 22, wherein the scFv light chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14.
Embodiment 24 includes the isolated polypeptide complex of embodiment 22, wherein the scFv light chain variable domain comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14.
Embodiment 25 includes an isolated polypeptide complex according to any of embodiments 1-24, wherein the scFv light chain variable domain comprises the amino acid sequence according to SEQ ID No. 11 or 14.
Embodiment 26 includes an isolated polypeptide complex according to any of embodiments 1-25, wherein the scFv comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 13 or 16.
Embodiment 27 includes an isolated polypeptide complex according to any of embodiments 1-25, wherein the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID No. 13 or 16.
Embodiment 28 includes the isolated polypeptide complex of embodiment 27, wherein the scFv comprises the amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID No. 13 or 16.
Embodiment 29 includes the isolated polypeptide complex of embodiment 27, wherein the scFv comprises the amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID No. 13 or 16 and has at least 80% sequence identity to at least 225 consecutive amino acid residues of SEQ ID No. 13 or 16.
Embodiment 30 includes an isolated polypeptide complex according to any of embodiments 1-29, wherein the scFv comprises an amino acid sequence according to SEQ ID No. 13 or 16.
Embodiment 31 includes an isolated polypeptide complex according to any of embodiments 1-30, wherein the linker connects the C-terminus of a to the N-terminus of B.
Embodiment 32 includes an isolated polypeptide complex according to any of embodiments 1-30, wherein the linker connects the N-terminus of a to the C-terminus of B.
Embodiment 33 includes an isolated polypeptide complex according to embodiment 31, wherein the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide.
Embodiment 34 includes the isolated polypeptide complex of embodiment 32, wherein the linker connects the N-terminus of a to the C-terminus of the Fab heavy chain polypeptide.
Embodiment 35 includes an isolated polypeptide complex according to embodiment 31, wherein the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide.
Embodiment 36 includes the isolated polypeptide complex of embodiment 32, wherein the linker connects the N-terminus of a to the C-terminus of the Fab light chain polypeptide.
Embodiment 37 includes an isolated polypeptide complex according to any of embodiments 1-32, wherein the linker connects the Fab light chain polypeptide to the scFv light chain variable domain.
Embodiment 38 includes an isolated polypeptide complex according to any of embodiments 1-32, wherein the linker connects the Fab light chain polypeptide to the scFv heavy chain variable domain.
Embodiment 39 includes an isolated polypeptide complex according to any of embodiments 1-32, wherein the linker connects the Fab heavy chain polypeptide to the scFv light chain variable domain.
Embodiment 40 includes an isolated polypeptide complex according to any of embodiments 1-32, wherein the linker connects the Fab heavy chain polypeptide to the scFv heavy chain variable domain.
Embodiment 41 includes the isolated polypeptide complex of embodiment 36, wherein the linker connects the Fab light chain polypeptide to the N-terminus of the scFv light chain variable domain.
Embodiment 42 includes an isolated polypeptide complex according to embodiment 35, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain.
Embodiment 43 includes an isolated polypeptide complex according to embodiment 34, wherein the linker connects the Fab light chain polypeptide to the N-terminus of the scFv heavy chain variable domain.
Embodiment 44 includes an isolated polypeptide complex according to embodiment 35, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain.
Embodiment 45 includes the isolated polypeptide complex of embodiment 34, wherein the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv light chain variable domain.
Embodiment 46 includes an isolated polypeptide complex according to embodiment 33, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain.
Embodiment 47 includes the isolated polypeptide complex of embodiment 35, wherein the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv heavy chain variable domain.
Embodiment 48 includes an isolated polypeptide complex according to embodiment 33, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain.
Embodiment 49 includes an isolated polypeptide complex according to any of embodiments 1-48, wherein the linker is at least 5 amino acids in length.
Embodiment 50 includes an isolated polypeptide complex according to any of embodiments 1-49, wherein the linker is no more than 30 amino acids in length.
Embodiment 51 includes an isolated polypeptide complex according to any of embodiments 1-50, wherein the linker is at least 5 amino acids and no more than 30 amino acids in length.
Embodiment 52 includes an isolated polypeptide complex according to any of embodiments 1-51, wherein the linker is 5 amino acids in length.
Embodiment 53 includes an isolated polypeptide complex according to any of embodiments 1-51, wherein the linker is 15 amino acids in length.
Embodiment 54 includes an isolated polypeptide complex according to any of embodiments 1-51, wherein the linker comprises the amino acid sequence of SEQ ID No. 32 ((GGGGSGGGGSGGGGS) or SEQ ID No. 33 (GGGGS).
Embodiment 55 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 34.
Embodiment 56 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34.
Embodiment 57 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34.
Embodiment 58 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 34.
Embodiment 59 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 34.
Embodiment 60 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 35.
Embodiment 61 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 35.
Embodiment 62 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35.
Embodiment 63 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35.
Embodiment 64 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 35.
Embodiment 65 includes the isolated polypeptide complex of embodiment 48, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 36.
Embodiment 66 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 36.
Embodiment 67 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36.
Embodiment 68 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36.
Embodiment 69 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 36.
Embodiment 70 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38.
Embodiment 71 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 72 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 73 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 74 includes the isolated polypeptide complex of embodiment 44, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 38.
Embodiment 75 includes the isolated polypeptide complex of embodiment 46, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 37.
Embodiment 76 includes the isolated polypeptide complex of embodiment 46, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37.
Embodiment 77 includes the isolated polypeptide complex of embodiment 46, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37.
Embodiment 78 includes the isolated polypeptide complex of embodiment 46, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37.
Embodiment 79 includes the isolated polypeptide complex of embodiment 46, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37.
Embodiment 80 includes the isolated polypeptide complex of embodiment 42, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38.
Embodiment 81 includes the isolated polypeptide complex of embodiment 42, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 82 includes the isolated polypeptide complex of embodiment 42, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 83 includes the isolated polypeptide complex of embodiment 42, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 84 includes the isolated polypeptide complex of embodiment 42, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID NO:31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 38.
Embodiment 85 includes an isolated polypeptide complex according to the formula: A-L-D ((formula II) wherein A comprises a single chain variable fragment (scFv) that binds to CD3, D comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA, and L comprises a linker that connects the C-terminus of A to the N-terminus of D.
Embodiment 86 includes the isolated polypeptide complex of embodiment 47, wherein the Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain.
Embodiment 87 includes the isolated polypeptide complex of embodiment 47 or 48, wherein the scFv comprises a scFv light chain variable domain and a scFv heavy chain variable domain.
Embodiment 88 includes an isolated polypeptide complex according to any one of embodiments 47-49, wherein the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide.
Embodiment 89 includes an isolated polypeptide complex according to any of embodiments 47-49, wherein the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide.
Embodiment 90 includes the isolated polypeptide complex of embodiment 89, wherein the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab heavy chain polypeptide.
Embodiment 91 includes an isolated polypeptide complex according to embodiment 89, wherein the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab light chain polypeptide.
Embodiment 92 includes the isolated polypeptide complex of embodiment 89, wherein the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab heavy chain polypeptide.
Embodiment 93 includes the isolated polypeptide complex of embodiment 89, wherein the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab light chain polypeptide.
Embodiment 94 includes an isolated polypeptide complex according to any of embodiments 47-55, wherein the Fab heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the Fab heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18, HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18, HC-CDR3: SEQ ID NO. 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3.
Embodiment 95 includes an isolated polypeptide complex according to any of embodiments 47-74, wherein the Fab light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 of the Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22; LC-CDR2: SEQ ID NO. 23; and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
Embodiment 96 includes an isolated polypeptide complex according to any of embodiments 47-75, wherein the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2 and HC-CDR3, wherein the HC-CDR1, HC-CDR2 and HC-CDR3 of the scFv heavy chain variable domain comprises: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3 SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein the CDR comprises 0-2 amino acid modifications in at least one of HC-CDR1, HC-CDR2 or HC-CDR 3.
Embodiment 97 includes an isolated polypeptide complex according to any of embodiments 47-76, wherein the scFv light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of the scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein the CDR comprises 0-2 amino acid modifications in at least one of LC-CDR1, LC-CDR2 or LC-CDR 3.
Embodiment 98 includes the isolated polypeptide complex of any of embodiments 48-59, wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 29 or 31.
Embodiment 99 includes an isolated polypeptide complex according to any of embodiments 48-59, wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 29 or 31.
Embodiment 100 includes an isolated polypeptide complex according to embodiment 99, wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 29 or 31.
Embodiment 101 includes the isolated polypeptide complex of embodiment 99, wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 29 or 31, and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 29 or 31.
Embodiment 102 includes an isolated polypeptide complex according to any of embodiments 86-101, wherein the Fab heavy chain polypeptide comprises an amino acid sequence according to SEQ ID No. 29 or 31.
Embodiment 103 includes the isolated polypeptide complex of any of embodiments 86-102, wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 28 or 30.
Embodiment 104 includes the isolated polypeptide complex of any of embodiments 86-102, wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID NO 28 or 30.
Embodiment 105 includes the isolated polypeptide complex of embodiment 104, wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 or 30.
Embodiment 106 includes the isolated polypeptide complex of embodiment 104, wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 or 30, and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28 or 30.
Embodiment 107 includes an isolated polypeptide complex according to any one of embodiments 85-106, wherein the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID No. 28 or 30.
Embodiment 108 includes the isolated polypeptide complex of any of embodiments 85-107, wherein the scFv heavy chain variable domain comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15.
Embodiment 109 includes an isolated polypeptide complex according to any one of embodiments 85-107, wherein the scFv heavy chain variable domain comprises the amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 12 or 15.
Embodiment 110 includes an isolated polypeptide complex according to embodiment 109, wherein the scFv heavy chain variable domain comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15.
Embodiment 111 includes the isolated polypeptide complex of embodiment 109, wherein the scFv heavy chain variable domain comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 12 or 15.
Embodiment 112 includes an isolated polypeptide complex according to any of embodiments 85-111, wherein the scFv heavy chain variable domain comprises an amino acid sequence according to SEQ ID No. 12 or 15.
Embodiment 113 includes an isolated polypeptide complex according to any of embodiments 85-112, wherein the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14.
Embodiment 114 includes an isolated polypeptide complex according to any of embodiments 85-113, wherein the scFv light chain variable domain comprises the amino acid sequence of at least 50 consecutive amino acid residues of SEQ ID No. 11 or 14.
Embodiment 115 includes the isolated polypeptide complex of any of embodiment 114, wherein the scFv light chain variable domain comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14.
Embodiment 116 includes the isolated polypeptide complex of embodiment 114, wherein the scFv light chain variable domain comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14 and has at least 80% sequence identity to at least 100 consecutive amino acid residues of SEQ ID No. 11 or 14.
Embodiment 117 includes an isolated polypeptide complex according to any of embodiments 85-116, wherein the scFv light chain variable domain comprises an amino acid sequence according to SEQ ID No. 11 or 14.
Embodiment 118 includes an isolated polypeptide complex according to any of embodiments 85-117, wherein the scFv comprises an amino acid sequence that has at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 13 or 16.
Embodiment 119 includes an isolated polypeptide complex according to any one of embodiments 85-117, wherein the scFv comprises an amino acid sequence of at least 150 consecutive amino acid residues of SEQ ID No. 13 or 16.
Embodiment 120 includes an isolated polypeptide complex according to embodiment 119, wherein the scFv comprises the amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID No. 13 or 16.
Embodiment 121 includes the isolated polypeptide complex of embodiment 119, wherein the scFv comprises the amino acid sequence of at least 225 consecutive amino acid residues of SEQ ID No. 13 or 16 and has at least 80% sequence identity to at least 225 consecutive amino acid residues of SEQ ID No. 13 or 16.
Embodiment 122 includes an isolated polypeptide complex according to any of embodiments 85-122, wherein the scFv comprises an amino acid sequence according to SEQ ID No. 13 or 16.
Embodiment 123 includes the isolated polypeptide complex of any of embodiments 85-122, wherein the linker is at least 5 amino acids in length.
Embodiment 124 includes the isolated polypeptide complex of embodiment 123, wherein the linker is no more than 30 amino acids in length.
Embodiment 125 includes the isolated polypeptide complex of embodiment 124, wherein the linker is at least 5 amino acids and no more than 30 amino acids in length.
Embodiment 126 includes the isolated polypeptide complex of embodiment 125, wherein the linker is 5 amino acids in length.
Embodiment 127 includes an isolated polypeptide complex according to embodiment 125, wherein the linker is 15 amino acids in length.
Embodiment 128 includes the isolated polypeptide complex of embodiment 125, wherein the linker comprises the amino acid sequence of SEQ ID NO. 32 (GGGGSGGGGSGGGGS) or SEQ ID NO. 33 (GGGGS).
Embodiment 129 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 34.
Embodiment 130 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 34.
Embodiment 131 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34.
Embodiment 132 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 28 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 34 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 34.
Embodiment 133 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 34.
Embodiment 134 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 35.
Embodiment 135 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 35.
Embodiment 136 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35.
Embodiment 137 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 35 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 35.
Embodiment 138 includes the isolated polypeptide complex of embodiment 92, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 35.
Embodiment 139 includes the isolated polypeptide complex of embodiment 93, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 36.
Embodiment 140 includes the isolated polypeptide complex of embodiment 93, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 36.
Embodiment 141 includes the isolated polypeptide complex of embodiment 93, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36.
Embodiment 142 includes the isolated polypeptide complex of embodiment 93, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 36 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 36.
Embodiment 143 includes the isolated polypeptide complex of embodiment 93, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID No. 36.
Embodiment 144 includes the isolated polypeptide complex of embodiment 90, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID NO: 37.
Embodiment 145 includes the isolated polypeptide complex of embodiment 90, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37.
Embodiment 146 includes the isolated polypeptide complex of embodiment 90, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37.
Embodiment 147 includes the isolated polypeptide complex of embodiment 90, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 37.
Embodiment 148 includes the isolated polypeptide complex of embodiment 90, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37.
Embodiment 149 comprises the isolated polypeptide complex of embodiment 91, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38.
Embodiment 150 includes the isolated polypeptide complex of embodiment 91, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 151 includes an isolated polypeptide complex according to embodiment 91, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 152 includes the isolated polypeptide complex of embodiment 91, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to at least 450 consecutive amino acid residues of SEQ ID No. 38.
Embodiment 153 includes an isolated polypeptide complex according to embodiment 91, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID No. 38.
Embodiment 154 includes a pharmaceutical composition comprising: the isolated polypeptide complex of any of embodiments 1-153; and a pharmaceutically acceptable excipient.
Embodiment 155 includes an isolated recombinant nucleic acid molecule encoding a polypeptide of the polypeptide complex of any of embodiments 1-153.
Embodiment 156 includes an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating lung cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 157 includes an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating breast cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 158 includes an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating cervical cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 159 includes the isolated polypeptide complex of any of embodiments 1-153 for use in a method of treating ovarian cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 161 comprises the isolated polypeptide complex of any of embodiments 1-153 for use in a method of treating colorectal cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 162 includes an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating pancreatic cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 163 comprises an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating gastric cancer, comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 164 includes an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating prostate cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex of any of embodiments 1-153.
Embodiment 165 includes an isolated polypeptide complex according to any of embodiments 1-153 for use in a method of treating metastatic castration resistant prostate cancer, the method comprising administering to a subject in need thereof the isolated polypeptide complex according to any of embodiments 1-153.
Examples
Example 1: PSMA polypeptide Complex binding (Ab-2 and Ab-3)
PSMA-and CD3 epsilon binding of PSMA polypeptide complexes Ab-2 and Ab-3 was assessed.
Ab-2 and Ab-3 comprise the sequences as set forth in Table 9. LC and HC refer to the light and heavy chain sequences of Fab. The scFv that binds to CD3 is linked to LC or HC of Fab.
TABLE 9 Ab-2 and Ab-3 sequences
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Ab-2 and Ab-3 binding was assessed using an enzyme-linked immunosorbent assay (ELISA). Biotinylated peptides were captured on neutravidin (neutravidin) coated plates. The secondary antibodies are used to detect the bound polypeptide complex. The data for Ab-2 and Ab-3 binding to PSMA can be seen in FIG. 2. The titration data for PSMA binding can be seen in tables 10-11 and fig. 3 and 4. Titration data for CD3 ε can be seen in tables 11-12 and FIGS. 5 and 6.
Table 10.
Table 11.
Table 12.
Table 13.
Example 2: in vitro efficacy of PSMA polypeptide complexes
The polypeptide complexes are then evaluated in functional in vitro tumor cell killing.
Briefly, cd8+ T cells and LNCaP tumor cells were seeded at a 3:1 ratio onto 96-well tissue culture treated flat bottom plates and allowed to adhere overnight. The next day, the medium and non-adherent cells were removed and replaced with fresh medium containing the indicated concentration of titrated polypeptide complex. T cell cytotoxicity and cell viability at 24 hours are shown in fig. 7, and T cell cytotoxicity and cell viability at 48 hours are shown in fig. 8.
Example 3: in vitro efficacy of PSMA polypeptide complexes Ab-4 and Ab-5
Polypeptide complexes were evaluated in a functional in vitro tumor cell killing assay using PSMA positive tumor cell line 22Rv1 or LNCaP. Tumor cell killing was measured using an xcelligent real-time cell analyzer from Agilent that relies on sensor impedance measurements (cell index) that increase as tumor cells adhere, spread and spread on the sensor surface. Also, as tumor cells are killed, the impedance decreases. 10,000 tumor cells were added per well and allowed to adhere overnight on a 96-well E plate. The next day, polypeptide complexes titrated in medium supplemented with human serum were added to wells along with 30,000 cd8+ T cells. Cell index measurements were obtained every 10 minutes for an additional 72 hours. The cell index was then multiplied by the number of hours (tumor cell growth kinetics) versus the concentration of the polypeptide complex, where the concentration required to reduce tumor growth by 50% (IC 50) was calculated using Graphpad Prism software (fig. 9, fig. 10, table 14).
Table 14.
Ab-4 Ab-5
22Rv1 IC50 pM 4409 4.8
LNCaP IC50 pM 86.0 0.7
Example 4: PSMA TCE pharmacokinetics in cynomolgus monkeys
The pharmacokinetics and exploratory safety of polypeptide molecules were evaluated in cynomolgus monkeys. Briefly, approximately 3kg body weight of cynomolgus monkeys were administered the polypeptides in IV boluses, and signs of adverse events were observed daily. No adverse events in life were observed. After administration, blood was collected in K2 EDTA tubes at specific time points and processed into plasma. The plasma was cryopreserved until analysis. The concentration of polypeptide molecules in plasma was measured via standard ELISA techniques relative to a reference standard diluted in control cyno plasma. Plasma concentration curves were fitted to a standard two-phase exponential equation representing the distribution and elimination phases (fig. 11). The pharmacokinetic fit enabled calculation of Cmax, half-life, distribution volume, clearance and 7 day area under the curve (AUC) as shown in table 15.
Table 15.
Ab-5 10μg/kg Unit (B)
C MAX 1.69 nM
t 1/2 2.17 Hr
V d 0.23 L
VSS 0.67 L
CL 24.49 mL/hr/kg
BW 3.00 kg
7-day AUC 141 nM·min
Example 5: PSMA TCE cytokine release
Cytokine release following administration of polypeptide molecules by IV bolus was assessed in cynomolgus monkeys. Briefly, approximately 3kg body weight of cynomolgus monkeys were administered the polypeptides in IV boluses, and signs of adverse events were observed daily. No adverse events in life were observed. After administration, blood was collected in K2 EDTA tubes at specific time points and processed into plasma. The plasma was cryopreserved until analysis. Plasma samples were analyzed for cytokines using the non-human primate cell count Th1/Th2 bead array kit from BD biosciences as per manufacturer's instructions. The levels of interferon gamma, tumor necrosis factor alpha, interleukin 6, interleukin 5, interleukin 4 and interleukin 2 in the plasma were calculated relative to the reference standard provided with the bead array kit (fig. 12A-12F, respectively).
Example 6: PSMA TCE liver enzyme in cynomolgus monkey
Systemic liver enzymes were evaluated in cynomolgus monkeys after administration of polypeptide molecules by IV bolus. Briefly, approximately 3kg body weight of cynomolgus monkeys were administered the polypeptides in IV boluses, and signs of adverse events were observed daily. No adverse events in life were observed. After administration, blood was collected in K2 EDTA tubes at specific time points and processed into plasma. The plasma was cryopreserved until analysis. Plasma samples were analyzed for the presence of the liver enzymes aspartate Aminotransferase (AST) and alanine Aminotransferase (ALT) as signs of potential liver toxicity. All time points tested post-dosing were kept within a constant range of AST and ALT levels, indicating no hepatotoxicity. AST and ALT were quantified according to the instructions provided in the commercial kit for Millipore. AST and ALT levels were calculated relative to positive control reference standards according to manufacturer's instructions (fig. 13).
Example 7: PSMA polypeptide Complex binding (Ab-4 and Ab-5)
PSMA and CD3 epsilon binding of PSMA polypeptide complexes Ab-4 and Ab-5 was assessed.
Ab-4 (SEQ ID NOS: 30 and 37) and Ab-5 (SEQ ID NOS: 38 and 31) include the sequences set forth in Table 8.
Ab-4 and Ab-5 binding was assessed using an enzyme-linked immunosorbent assay (ELISA). Biotinylated peptides were captured on neutravidin coated plates. The secondary antibodies are used to detect the bound polypeptide complex. Data for Ab-4 and Ab-5 binding to PSMA (Ab-4 EC50=2.8 nM; ab-5 EC50=2.0 nM) are shown in FIG. 14. Data for Ab-4 and Ab-5 binding to CD3 (Ab-4 EC50=0.1 nM; ab-5 EC50=0.17 nM) are shown in FIG. 15.
Sequence listing
<110> Jianukesi treatment Co
<120> antibodies targeting PSMA and CD3 and uses thereof
<130> 52426-723.601
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<150> 63/188,855
<151> 2021-05-14
<150> 63/092,226
<151> 2020-10-15
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Peptides
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Gly Phe Thr Phe Asn Lys Tyr Ala
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Val Arg His Gly Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr
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Thr Gly Ala Val Thr Thr Ser Asn Tyr
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Ala Leu Trp Tyr Ser Asn Leu Trp Val
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Polypeptides
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Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Gly Ser Ser Thr Gly Ala Val Thr Ser Gly
20 25 30
Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Lys Phe Leu Ala Pro Gly Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Val Leu Trp Tyr Ser Asn
85 90 95
Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 12
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Polypeptides
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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
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Polypeptides
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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Thr Val Val
130 135 140
Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu
145 150 155 160
Thr Cys Gly Ser Ser Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn
165 170 175
Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly
180 185 190
Thr Lys Phe Leu Ala Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu
195 200 205
Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp
210 215 220
Glu Ala Glu Tyr Tyr Cys Val Leu Trp Tyr Ser Asn Arg Trp Val Phe
225 230 235 240
Gly Gly Gly Thr Lys Leu Thr Val Leu
245
<210> 14
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Polypeptides
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Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 15
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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 16
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Polypeptides
<400> 16
Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
115 120 125
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu
130 135 140
Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala Met Asn Trp
145 150 155 160
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg
165 170 175
Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp
180 185 190
Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln
195 200 205
Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg
210 215 220
His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr Trp Gly
225 230 235 240
Gln Gly Thr Leu Val Thr Val Ser Ser
245
<210> 17
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Gly Phe Thr Phe Ser Asn Tyr Val
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Ile Trp Tyr Asp Gly Ser Asn Lys
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Ala Gly Gly Tyr Asn Trp Asn Tyr Glu Tyr His Tyr Tyr Gly Met Asp
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Val
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Gly Phe Ala Phe Ser Arg Tyr Gly
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Ala Arg Gly Gly Asp Phe Leu Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val
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Gln Gly Ile Thr Asn Tyr
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Ala Ala
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Gln Gln Tyr Asn Ser Tyr Pro Ile Thr
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Gln Gly Ile Ser Asn Tyr
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<211> 2
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Peptides
<400> 26
Glu Ala
1
<210> 27
<211> 9
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Peptides
<400> 27
Gln Asn Tyr Asn Ser Ala Pro Phe Thr
1 5
<210> 28
<211> 214
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 28
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Thr Asn Tyr
20 25 30
Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile
35 40 45
Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Lys Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Ile
85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 29
<211> 227
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 29
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Val Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Ile Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Gly Gly Tyr Asn Trp Asn Tyr Glu Tyr His Tyr Tyr Gly Met Asp
100 105 110
Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125
Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly
130 135 140
Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
145 150 155 160
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175
Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
180 185 190
Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn
195 200 205
Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro
210 215 220
Lys Ser Cys
225
<210> 30
<211> 214
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 30
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Thr Gly Lys Val Pro Lys Phe Leu Ile
35 40 45
Tyr Glu Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Gly Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Asn Tyr Asn Ser Ala Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 31
<211> 226
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 31
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Arg Tyr
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Gln Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Asp Phe Leu Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys
225
<210> 32
<211> 15
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Peptides
<400> 32
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 33
<211> 5
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Peptides
<400> 33
Gly Gly Gly Gly Ser
1 5
<210> 34
<211> 481
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 34
Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
115 120 125
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu
130 135 140
Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala Met Asn Trp
145 150 155 160
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg
165 170 175
Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp
180 185 190
Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln
195 200 205
Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg
210 215 220
His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr Trp Gly
225 230 235 240
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gln Val
245 250 255
Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu
260 265 270
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr Val Met
275 280 285
His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ile
290 295 300
Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly
305 310 315 320
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln
325 330 335
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Gly
340 345 350
Gly Tyr Asn Trp Asn Tyr Glu Tyr His Tyr Tyr Gly Met Asp Val Trp
355 360 365
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
370 375 380
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
385 390 395 400
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
405 410 415
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
420 425 430
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
435 440 445
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
450 455 460
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
465 470 475 480
Cys
<210> 35
<211> 480
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 35
Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
115 120 125
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu
130 135 140
Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala Met Asn Trp
145 150 155 160
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg
165 170 175
Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp
180 185 190
Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln
195 200 205
Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg
210 215 220
His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr Trp Gly
225 230 235 240
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gln Val
245 250 255
Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu
260 265 270
Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Arg Tyr Gly Met
275 280 285
His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val
290 295 300
Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly
305 310 315 320
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Gln Tyr Leu Gln
325 330 335
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
340 345 350
Gly Gly Asp Phe Leu Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly
355 360 365
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
370 375 380
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
385 390 395 400
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
405 410 415
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
420 425 430
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
435 440 445
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
450 455 460
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
465 470 475 480
<210> 36
<211> 468
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 36
Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
115 120 125
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu
130 135 140
Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala Met Asn Trp
145 150 155 160
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg
165 170 175
Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp
180 185 190
Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln
195 200 205
Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg
210 215 220
His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr Trp Gly
225 230 235 240
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile
245 250 255
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg
260 265 270
Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr Leu Ala
275 280 285
Trp Tyr Gln Gln Lys Thr Gly Lys Val Pro Lys Phe Leu Ile Tyr Glu
290 295 300
Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Gly Gly
305 310 315 320
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp
325 330 335
Val Ala Thr Tyr Tyr Cys Gln Asn Tyr Asn Ser Ala Pro Phe Thr Phe
340 345 350
Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala Pro Ser
355 360 365
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
370 375 380
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
385 390 395 400
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
405 410 415
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
420 425 430
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
435 440 445
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
450 455 460
Arg Gly Glu Cys
465
<210> 37
<211> 480
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 37
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Thr Val Val
130 135 140
Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu
145 150 155 160
Thr Cys Gly Ser Ser Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn
165 170 175
Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly
180 185 190
Thr Lys Phe Leu Ala Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu
195 200 205
Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp
210 215 220
Glu Ala Glu Tyr Tyr Cys Val Leu Trp Tyr Ser Asn Arg Trp Val Phe
225 230 235 240
Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Gln Val
245 250 255
Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu
260 265 270
Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Arg Tyr Gly Met
275 280 285
His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val
290 295 300
Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly
305 310 315 320
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Gln Tyr Leu Gln
325 330 335
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
340 345 350
Gly Gly Asp Phe Leu Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly
355 360 365
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
370 375 380
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
385 390 395 400
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
405 410 415
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
420 425 430
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
435 440 445
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
450 455 460
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
465 470 475 480
<210> 38
<211> 468
<212> PRT
<213> artificial sequence
<220>
<223> description of artificial sequence synthetic
Polypeptides
<400> 38
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Thr Val Val
130 135 140
Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu
145 150 155 160
Thr Cys Gly Ser Ser Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn
165 170 175
Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly
180 185 190
Thr Lys Phe Leu Ala Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu
195 200 205
Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp
210 215 220
Glu Ala Glu Tyr Tyr Cys Val Leu Trp Tyr Ser Asn Arg Trp Val Phe
225 230 235 240
Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Asp Ile
245 250 255
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg
260 265 270
Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr Leu Ala
275 280 285
Trp Tyr Gln Gln Lys Thr Gly Lys Val Pro Lys Phe Leu Ile Tyr Glu
290 295 300
Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Gly Gly
305 310 315 320
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp
325 330 335
Val Ala Thr Tyr Tyr Cys Gln Asn Tyr Asn Ser Ala Pro Phe Thr Phe
340 345 350
Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala Pro Ser
355 360 365
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
370 375 380
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
385 390 395 400
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
405 410 415
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
420 425 430
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
435 440 445
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
450 455 460
Arg Gly Glu Cys
465

Claims (72)

1. An isolated polypeptide complex according to the formula:
A-L-B
(formula I)
Wherein the method comprises the steps of
A comprises a single chain variable fragment (scFv) that binds to CD 3;
b comprises an antigen binding fragment (Fab) or Fab 'that binds to PSMA, wherein the Fab or Fab' comprises a Fab light chain polypeptide comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain comprising a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the Fab heavy chain variable domain comprise HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18 and HC-CDR3: 21, and wherein the CDR comprises 0-2 amino acid modifications in at least one of the HC-CDR1, HC-CDR2 or HC-CDR 3; and is also provided with
L comprises a linker connecting a to B.
2. The isolated polypeptide complex of claim 1, wherein the Fab light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2, and LC-CDR3, wherein said LC-CDR1, said LC-CDR2, and said LC-CDR3 of said Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22, LC-CDR2: SEQ ID NO. 23 and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said LC-CDR1, LC-CDR2 or LC-CDR 3.
3. The isolated polypeptide complex of claim 1, wherein the scFv comprises a scFv light chain variable domain and a scFv heavy chain variable domain.
4. The isolated polypeptide complex of claim 3, wherein the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the scFv heavy chain variable domain comprise HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said HC-CDR1, HC-CDR2 or HC-CDR 3.
5. The isolated polypeptide complex of claim 4, wherein the scFv light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2, and LC-CDR3, wherein said LC-CDR1, said LC-CDR2, and said LC-CDR3 of said scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said LC-CDR1, LC-CDR2 or LC-CDR 3.
6. The isolated polypeptide complex of claim 1, wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 29 or 31.
7. The isolated polypeptide complex of claim 1, wherein the Fab heavy chain polypeptide comprises an amino acid sequence according to SEQ ID No. 29 or 31.
8. The isolated polypeptide complex of claim 1, wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID NO 28 or 30.
9. The isolated polypeptide complex of claim 1, wherein the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID No. 28 or 30.
10. The isolated polypeptide complex of claim 3, wherein the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 12 or 15.
11. The isolated polypeptide complex of claim 3, wherein the scFv heavy chain variable domain comprises an amino acid sequence according to SEQ ID No. 12 or 15.
12. The isolated polypeptide complex of claim 3, wherein the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID No. 11 or 14.
13. The isolated polypeptide complex of claim 3, wherein the scFv light chain variable domain comprises an amino acid sequence according to SEQ ID No. 11 or 14.
14. The isolated polypeptide complex of claim 1, wherein the scFv comprises an amino acid sequence having at least 80% sequence identity to an amino acid sequence according to SEQ ID NO 13 or 16.
15. The isolated polypeptide complex of claim 1, wherein the scFv comprises an amino acid sequence according to SEQ ID No. 13 or 16.
16. The isolated polypeptide complex of claim 1, wherein the linker connects the C-terminus of a to the N-terminus of B.
17. The isolated polypeptide complex of claim 1, wherein the linker connects the N-terminus of a to the C-terminus of B.
18. The isolated polypeptide complex of claim 1, wherein the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide.
19. The isolated polypeptide complex of claim 2, wherein the linker connects the N-terminus of a to the C-terminus of the Fab heavy chain polypeptide.
20. The isolated polypeptide complex of claim 1, wherein the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide.
21. The isolated polypeptide complex of claim 2, wherein the linker connects the N-terminus of a to the C-terminus of the Fab light chain polypeptide.
22. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the scFv light chain variable domain.
23. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the scFv heavy chain variable domain.
24. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the scFv light chain variable domain.
25. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the scFv heavy chain variable domain.
26. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the N-terminus of the scFv light chain variable domain.
27. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain.
28. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the N-terminus of the scFv heavy chain variable domain.
29. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain.
30. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv light chain variable domain.
31. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain.
32. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the N-terminus of the scFv heavy chain variable domain.
33. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain.
34. The isolated polypeptide complex of claim 1, wherein the linker comprises the amino acid sequence of SEQ ID No. 32 (GGGGSGGGGSGGGGS) or SEQ ID No. 33 (GGGGS).
35. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 28, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 34.
36. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:28 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 34.
37. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 35.
38. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence of SEQ ID NO: 35.
39. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 36.
40. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID NO:31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 36.
41. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38.
42. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv heavy chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID NO:31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 38.
43. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 37.
44. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO:30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 37.
45. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38.
46. The isolated polypeptide complex of claim 3, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID NO:31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 38.
47. An isolated polypeptide complex according to the formula:
A-L-D
(formula II)
Wherein the method comprises the steps of
A comprises a single chain variable fragment (scFv) that binds to CD 3;
d comprises an antigen binding fragment (Fab) or Fab' that binds to PSMA; and is also provided with
L comprises a linker connecting the C-terminus of A to the N-terminus of D.
48. The isolated polypeptide complex of claim 47, wherein the Fab or Fab' comprises a Fab light chain polypeptide chain comprising a Fab light chain variable domain and a Fab heavy chain polypeptide comprising a Fab heavy chain variable domain.
49. The isolated polypeptide complex of claim 48, wherein the scFv comprises a scFv light chain variable domain and a scFv heavy chain variable domain.
50. The isolated polypeptide complex of claim 49, wherein the linker connects the C-terminus of a to the N-terminus of the Fab heavy chain polypeptide.
51. The isolated polypeptide complex of claim 49, wherein the linker connects the C-terminus of a to the N-terminus of the Fab light chain polypeptide.
52. The isolated polypeptide complex of claim 49, wherein the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab heavy chain polypeptide.
53. The isolated polypeptide complex of claim 49, wherein the linker connects the C-terminus of the scFv light chain variable domain to the N-terminus of the Fab light chain polypeptide.
54. The isolated polypeptide complex of claim 49, wherein the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab heavy chain polypeptide.
55. The isolated polypeptide complex of claim 49, wherein the linker connects the C-terminus of the scFv heavy chain variable domain to the N-terminus of the Fab light chain polypeptide.
56. The isolated polypeptide complex of claim 49, wherein the Fab heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the Fab heavy chain variable domain comprise HC-CDR1: SEQ ID NO. 17, HC-CDR2: SEQ ID NO. 18, HC-CDR3: SEQ ID NO. 19; or HC-CDR1: SEQ ID NO. 20, HC-CDR2: SEQ ID NO. 18, HC-CDR3: SEQ ID NO. 21, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said HC-CDR1, HC-CDR2 or HC-CDR 3.
57. The isolated polypeptide complex of claim 49, wherein the Fab light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2, and LC-CDR3, wherein said LC-CDR1, said LC-CDR2, and said LC-CDR3 of said Fab light chain variable domain comprise LC-CDR1: SEQ ID NO. 22; LC-CDR2: SEQ ID NO. 23; and LC-CDR3: SEQ ID NO. 24; or LC-CDR1: SEQ ID NO. 25, LC-CDR2: SEQ ID NO 26 and LC-CDR3: SEQ ID NO. 27, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said LC-CDR1, LC-CDR2 or LC-CDR 3.
58. The isolated polypeptide complex of claim 49, wherein the scFv heavy chain variable domain comprises a Complementarity Determining Region (CDR): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the scFv heavy chain variable domain comprise: HC-CDR1: SEQ ID NO. 1, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 3; or HC-CDR1: SEQ ID NO. 4, HC-CDR2: SEQ ID NO. 2 and HC-CDR3: SEQ ID NO. 5, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said HC-CDR1, HC-CDR2 or HC-CDR 3.
59. The isolated polypeptide complex of claim 49, wherein the scFv light chain variable domain comprises a Complementarity Determining Region (CDR): LC-CDR1, LC-CDR2, and LC-CDR3, wherein said LC-CDR1, said LC-CDR2, and said LC-CDR3 of said scFv light chain variable domain comprises LC-CDR1: SEQ ID NO. 6, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 8; or LC-CDR1: SEQ ID NO. 9, LC-CDR2: SEQ ID NO. 7 and LC-CDR3: SEQ ID NO. 10, and wherein said CDR comprises 0-2 amino acid modifications in at least one of said LC-CDR1, LC-CDR2 or LC-CDR 3.
60. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 37.
61. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 37.
62. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37.
63. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 30 and has at least 80% sequence identity to the at least 200 consecutive amino acid residues of SEQ ID No. 30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 37 and has at least 80% sequence identity to the at least 450 consecutive amino acid residues of SEQ ID No. 37.
64. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab heavy chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab light chain polypeptide comprises an amino acid sequence according to SEQ ID NO:30, and the amino acid sequence of the Fab heavy chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence of SEQ ID NO: 37.
65. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence according to SEQ ID No. 38.
66. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 100 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence of at least 400 consecutive amino acid residues of SEQ ID No. 38.
67. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises an amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises an amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38.
68. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence of at least 200 consecutive amino acid residues of SEQ ID No. 31 and has at least 80% sequence identity to the at least 200 consecutive amino acid residues of SEQ ID No. 31, and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of at least 450 consecutive amino acid residues of SEQ ID No. 38 and has at least 80% sequence identity to the at least 450 consecutive amino acid residues of SEQ ID No. 38.
69. The isolated polypeptide complex of claim 49, wherein the linker connects the Fab light chain polypeptide to the C-terminus of the scFv light chain variable domain, and wherein the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID NO:31 and the amino acid sequence of the Fab light chain polypeptide connected to the C-terminus of the scFv light chain variable domain comprises the amino acid sequence of SEQ ID NO: 38.
70. A pharmaceutical composition comprising:
(i) The isolated polypeptide complex of any one of claims 1-69; and
(ii) Pharmaceutically acceptable excipients.
71. An isolated recombinant nucleic acid molecule encoding a polypeptide of the polypeptide complex of any one of claims 1-69.
72. A method of treating metastatic castration-resistant prostate cancer in a subject in need thereof, the method comprising administering to the subject the polypeptide complex of any one of claims 1-69.
CN202180084823.5A 2020-10-15 2021-10-14 Antibodies targeting PSMA and CD3 and uses thereof Pending CN116997573A (en)

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US202163188855P 2021-05-14 2021-05-14
US63/188,855 2021-05-14
PCT/US2021/054948 WO2022081822A1 (en) 2020-10-15 2021-10-14 Antibodies targeting psma and cd3 and uses thereof

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