Serum albumin binding proteins containing immunoglobulin single variable domains
This invention provides, and in certain specific but non-limiting aspects relates to: assays that can be used to predict whether a given ISV will be subject to protein interference and/or give rise to an (aspecific) signal in such an assay (such as in an ADA immunoassay; methods for modifying and/or improving ISV's to as to remove or reduce their tendency to give rise to such protein interference or such a signal; modifications that can be introduced into an ISV that remove or reduce its tendency to give rise to such protein interference or such a signal; ISV's that have been specifically selected (for example, using the assay(s) described herein) to have no or low(er)/reduced tendency to give rise to such protein interference or such a signal; modified and/or improved ISV's that have no or a low(er)/reduced tendency to give rise to such protein interference or such a signal.
1. A protein or polypeptide comprising an immunoglobulin single variable domain (ISV) at its C-terminal end, wherein said ISV is a VHH, humanized VHH or camelized VH, which ISV has a C-terminal end of the sequence VTVSS(X) n (SEQ ID NO: 34), in which each X is an amino acid residue that is independently chosen and wherein:
n=1 or 2 in which each X=Ala or Gly; or
n=1 or 2 in which each X=Ala; or
n=1, 2 or 3 in which each X=Gly; or
n=2 or 3 in which at least one X=Ala or Gly, wherein when n=3, at least one X is not Ala; or
n=2 or 3 in which all but one X=Ala or Gly,
and which protein or polypeptide includes a serum albumin binding peptide or serum albumin binding domain linked to the ISV directly or via one or more linkers.
2. The protein or polypeptide according to claim 1 , in which the protein or polypeptide comprises a serum albumin binding domain that is a serum albumin binding ISV, wherein said serum albumin binding ISV is a VHH, humanized VHH or camelized VH.
3. The protein or polypeptide according to claim 2 , in which said serum albumin binding ISV is a VHH or a humanized VHH that is Alb-1 or a humanized version of Alb-1.
4. The protein or polypeptide according to claim 3 , in which said humanized version of Alb-1 is Alb-8.
5. The protein or polypeptide according to claim 1 , in which n=1 or n=2.
6. The protein or polypeptide according to claim 1 , in which:
n=2 or 3 and at least one X=Ala or Gly, wherein when n=3, at least X is not Ala; or
n=2 or 3 and all but one X=Ala or Gly.
7. The protein or polypeptide according to claim 6 , in which the remaining amino acid residue X is independently chosen from Val, Leu and/or Ile.
8. The protein or polypeptide according to claim 1 , in which:
n=1 or 2 in which each X=Ala or Gly; or
n=1 or 2 in which each X=Ala; or
n=1, 2 or 3 in which each X=Gly.
9. The protein or polypeptide according to claim 1 , in which X is not cysteine.
10. A pharmaceutical composition comprising a protein or polypeptide according to claim 1 , and at least one suitable carrier, diluent or excipient.
11. A protein or polypeptide comprising an immunoglobulin single variable domain (ISV) at its C-terminal end, wherein said ISV is a VHH, humanized VHH or camelized VH, which ISV has a C-terminal end of the sequence VTVSS(X) n (SEQ ID NO: 34), in which n is 1, 2, 3, or 5, and in which each X is an amino acid residue that is independently chosen, with the proviso that X is not cysteine, wherein when n=3, at least one amino acid residue X is not alanine, and which protein or polypeptide includes a serum albumin binding peptide or serum albumin binding domain linked to the ISV directly or via one or more linkers.
12. The protein or polypeptide according to claim 11 , in which each X is a naturally occurring amino acid.
13. The protein or polypeptide according to claim 11 , in which the protein or polypeptide comprises a serum albumin binding domain that is a serum albumin binding ISV, wherein said serum albumin binding ISV is a VHH, humanized VHH or camelized VH.
14. The protein or polypeptide according to claim 13 , in which said serum albumin binding ISV is a VHH or a humanized VHH that is Alb-1 or a humanized version of Alb-1.
15. The protein or polypeptide according to claim 14 , in which said humanized version of Alb-1 is Alb-8.
16. A pharmaceutical composition comprising a protein or polypeptide according to claim 11 , and at least one suitable carrier, diluent or excipient.
17. A protein or polypeptide comprising an immunoglobulin single variable domain (ISV) at its C-terminal end, wherein said ISV is a VHH, humanized VHH or camelized VH, which ISV has a C-terminal end of the sequence VTVSS(X) n (SEQ ID NO: 34), in which n is 1 to 5, in which each X is independently chosen from the group consisting of alanine (A), glycine (G), valine (V), leucine (L) or isoleucine (I), wherein when n=3, at least one amino acid residue X is not alanine, and which protein or polypeptide includes a serum albumin binding peptide or serum albumin binding domain linked to the ISV directly or via one or more linkers.
18. The protein or polypeptide according to claim 17 , in which the protein or polypeptide comprises a serum albumin binding domain that is a serum albumin binding ISV, wherein said serum albumin binding ISV is a VHH, humanized VHH or camelized VH.
19. The protein or polypeptide according to claim 18 , in which said serum albumin binding ISV is a VHH or a humanized VHH that is Alb-1 or a humanized version of Alb-1.
20. The protein or polypeptide according to claim 19 , in which said humanized version of Alb-1 is Alb-8.
21. A pharmaceutical composition, comprising a protein or polypeptide according to claim 17 , and at least one suitable carrier, diluent or excipient.
22. A protein or polypeptide comprising an immunoglobulin single variable domain (ISV) at its C-terminal end that binds a therapeutic target, wherein said ISV is a VHH, humanized VHH or camelized VH, which ISV has a C-terminal end of the sequence VTVSS(X) n (SEQ ID NO: 34), in which each X is an amino acid residue that is independently chosen and wherein:
n=1, 2 or 3 in which each X=Ala or Gly; or
n=1, 2 or 3 in which each X=Ala; or
n=1, 2 or 3 in which each X=Gly; or
n=2 or 3 in which at least one X=Ala or Gly; or
n=2 or 3 in which all but one X=Ala or Gly,
and which protein or polypeptide includes a serum albumin binding peptide or serum albumin binding domain linked to the ISV directly or via one or more linkers.
23. The protein or polypeptide according to claim 22 , in which the remaining amino acid residue X is independently chosen from Val, Leu and/or Ile.
24. The protein or polypeptide according to claim 22 , in which X is not cysteine.
25. The protein or polypeptide according to claim 22 , in which the protein or polypeptide comprises a serum albumin binding domain that is a serum albumin binding ISV, wherein said serum albumin binding ISV is a VHH, humanized VHH or camelized VH.
26. The protein or polypeptide according to claim 25 , in which said serum albumin binding ISV is a VHH or a humanized VHH that is Alb-1 or a humanized version of Alb-1.
27. The protein or polypeptide according to claim 26 , in which said humanized version of Alb-1 is Alb-8.
28. A pharmaceutical composition comprising a protein or polypeptide according to claim 22 , and at least one suitable carrier, diluent or excipient.
29. A protein or polypeptide comprising an immunoglobulin single variable domain (ISV) at its C-terminal end that binds a therapeutic target, wherein said ISV is a VHH, humanized VHH or camelized VH, which ISV has a C-terminal end of the sequence VTVSS(X) n (SEQ ID NO: 34), in which n is 1 to 5, and in which each X is an amino acid residue that is independently chosen, with the proviso that X is not cysteine, and which protein or polypeptide includes a serum albumin binding peptide or serum albumin binding domain linked to the ISV directly or via one or more linkers.
30. The protein or polypeptide according to claim 29 , in which each X is a naturally occurring amino acid.
31. The protein or polypeptide according to claim 29 , in which each X is chosen from the group consisting of alanine (A), glycine (G), valine (V), leucine (L) or isoleucine (I).
32. The protein or polypeptide according to claim 29 , in which the protein or polypeptide comprises a serum albumin binding domain that is a serum albumin binding ISV, wherein said serum albumin binding ISV is a VHH, humanized VHH or camelized VH.
33. The protein or polypeptide according to claim 32 , in which said serum albumin binding ISV is a VHH or a humanized VHH that is Alb-1 or a humanized version of Alb-1.
34. The protein or polypeptide according to claim 33 , in which said humanized version of Alb-1 is Alb-8.
35. A pharmaceutical composition comprising a protein or polypeptide according to claim 29 , and at least one suitable carrier, diluent or excipient.