Cross-species-specific single domain bispecific single chain antibody
The present invention relates to a bispecific single chain antibody molecule comprising a first binding domain consisting of one antibody variable domain capable of binding to an epitope of the human and non-chimpanzee primate CD3 epsilon chain, wherein the epitope is part of an amino acid sequence comprised in the group consisting of SEQ ID NOs. 2, 4, 6, and 8, and a second binding domain capable of binding to an epitope of a human and a non-chimpanzee primate tumor target antigen. The invention further relates to a bispecific single chain antibody molecule comprising a first binding domain capable of binding to an epitope of human and non-chimpanzee primate CD3ε (epsilon) chain, wherein the epitope is part of an amino acid sequence comprised in the group consisting of SEQ ID NOs. 2, 4, 6, and 8, and a second binding domain consisting of one antibody variable domain capable of binding to an epitope of a human and a non-chimpanzee primate tumor target antigen. The invention also provides nucleic acids encoding said bispecific single chain antibody molecule as well as vectors and host cells and a process for its production. The invention further relates to pharmaceutical compositions comprising said bispecific single chain antibody molecule and medical uses of said bispecific single chain antibody molecule.
1. A nucleic acid comprising a polynucleotide encoding a bispecific single chain antibody molecule comprising a first binding domain which binds to an epitope of a human CD3 epsilon (CD3ε) chain, wherein the epitope is part of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, or 8, and comprises at least the amino acid sequence Gln-Asp-Gly-Asn-Glu (QDGNE; SEQ ID NO: 430), and a second binding domain comprising an antibody variable domain that is cross-species specific and binds to an epitope of a human and a non-chimpanzee primate tumor target antigen, wherein the first binding domain is a VHH domain and comprises CDR 1 of SEQ ID NO: 398, CDR 2 of SEQ ID NO: 399 and CDR 3 of SEQ ID. NO: 400.
2. The nucleic acid of claim 1 , wherein the first binding domain further binds a Callithrix jacchus, Sanguinis oedipus or Saimiri sciureus CD3ε chain.
3. The nucleic acid of claim 1 , wherein at least one of said first or second binding domain is CDR-grafted, humanized or human.
4. The nucleic acid of claim 1 , wherein the first binding domain comprises an antibody variable domain comprising:
(a) the amino acid sequence of SEQ ID NO. 397;
(b) an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 401;
(c) an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 397 or the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 401; or
(d) an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 397 or the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 401.
5. The nucleic acid of claim 1 , wherein the antibody variable domain of the second binding domain comprises CDR1-3 selected from the group consisting of:
(a) CDR 1 of SEQ ID NO: 376, CDR 2 of SEQ ID NO. 377 and CDR 3 of SEQ ID NO. 378; and
(b) CDR 1 of SEQ ID NO: 387, CDR 2 of SEQ ID NO. 388 and CDR 3 of SEQ ID NO. 389.
6. The nucleic acid of claim 1 , wherein the second binding domain comprises an antibody variable domain as shown in SEQ ID NO: 375 or 386 or an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 375 or 386.
7. The nucleic acid of claim 1 , wherein the bispecific single chain antibody molecule comprises a sequence selected from:
(a) the amino acid sequence of SEQ ID NO: 380, 382, 384, 391, 393 or 395;
(b) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 381, 383, 385, 392, 394 or 396; and
(c) an amino acid sequence at least 85% identical to the amino acid sequence of (a) or (b).
8. The nucleic acid of claim 1 , wherein the second binding domain comprises one antibody variable domain or two antibody variable domains.
9. The nucleic acid of claim 1 , wherein the second binding domain comprises a VHH domain.
10. The nucleic acid of claim 1 , wherein the tumor target antigen is EGFR, CD44v6 or CD30.
11. A nucleic acid comprising a polynucleotide encoding a bispecific single chain antibody molecule comprising a first binding domain, which binds to an epitope of a human CD3 epsilon (CD3ε) chain, wherein the epitope is part of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, or 8, and a second binding domain consisting of one antibody variable domain that is cross-species specific and binds to an epitope of a human and a non-chimpanzee primate tumor target antigen, wherein the first binding domain is a VHH domain and comprises CDR 1 of SEQ ID NO: 398, CDR 2 of SEQ ID NO: 399 and CDR 3 of SEQ ID. NO: 400.
12. The nucleic acid of claim 11 , wherein the first binding domain further binds a non-chimpanzee primate CD3ε chain.
13. The nucleic acid of claim 11 , wherein the first binding domain further binds a Callithrix jacchus, Sanguinis oedipus or Saimiri sciureus CD3ε chain.
14. The nucleic acid of claim 11 , wherein at least one of said first or second binding domain is CDR-grafted, humanized or human.
15. The nucleic acid of claim 11 , wherein the first binding domain comprises an antibody variable domain comprising:
(a) the amino acid sequence of SEQ ID NO. 397;
(b) an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 401;
(c) an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 397 or the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 401; or
(d) an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 397 or the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 401.
16. The nucleic acid of claim 11 , wherein the antibody variable domain of the second binding domain comprises CDR1-3 selected from the group consisting of:
(a) CDR 1 of SEQ ID NO: 376, CDR 2 of SEQ ID NO. 377 and CDR 3 of SEQ ID NO. 378; and
(b) CDR 1 of SEQ ID NO: 387, CDR 2 of SEQ ID NO. 388 and CDR 3 of SEQ ID NO. 389.
17. The nucleic acid of claim 11 , wherein the second binding domain comprises an antibody variable domain as shown in SEQ ID NO: 375 or 386 or an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 375 or 386.
18. The nucleic acid of claim 11 , wherein the bispecific single chain antibody molecule comprises a sequence selected from:
(a) the amino acid sequence of SEQ ID NO: 380, 382, 384, 391, 393 or 395;
(b) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 381, 383, 385, 392, 394 or 396; and
(c) an amino acid sequence at least 85% identical to the amino acid sequence of (a) or (b).
19. The nucleic acid of claim 11 , wherein the second binding domain comprises one antibody variable domain or two antibody variable domains.
20. The nucleic acid of claim 11 , wherein the second binding domain comprises a VHH domain.
21. The nucleic acid of claim 11 , wherein the tumor target antigen is EGFR, CD44v6 or CD30.
22. A vector comprising the nucleic acid of claim 1 .
23. A vector comprising the nucleic acid of claim 11 .
24. A host cell comprising the vector of claim 22 .
25. A host cell comprising the vector of claim 23 .
26. A process for producing a bispecific single chain antibody molecule comprising a first binding domain which binds to an epitope of a human CD3 epsilon (CD3ε) chain, wherein the epitope is part of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, or 8, and comprises at least the amino acid sequence Gln-Asp-Gly-Asn-Glu (QDGNE; SEQ ID NO: 430), and a second binding domain comprising an antibody variable domain that is cross-species specific and binds to an epitope of a human and a non-chimpanzee primate tumor target antigen, wherein the first binding domain is a VHH domain and comprises CDR 1 of SEQ ID NO: 398, CDR 2 of SEQ ID NO: 399 and CDR 3 of SEQ ID. NO: 400, the process comprising culturing the host cell of claim 24 under conditions allowing the expression of the bispecific single chain antibody molecule and recovering the molecule from the culture.
27. A process for producing a bispecific single chain antibody molecule comprising a first binding domain, which binds to an epitope of a human CD3 epsilon (CD3ε) chain, wherein the epitope is part of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, or 8, and a second binding domain consisting of one antibody variable domain that is cross-species specific and binds to an epitope of a human and a non-chimpanzee primate tumor target antigen, wherein the first binding domain is a VHH domain and comprises CDR 1 of SEQ ID NO: 398, CDR 2 of SEQ ID NO: 399 and CDR 3 of SEQ ID. NO: 400, the process comprising culturing the host cell of claim 25 under conditions allowing the expression of the bispecific single chain antibody molecule and recovering the molecule from the culture.