Albumin variants
The present invention relates to variants of a parent albumin, the variants having altered plasma half-life compared with the parent albumin. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.
1. A method of altering the binding activity of a polypeptide, which is a variant of albumin, a fragment thereof or a fusion polypeptide comprising said variant albumin or fragment thereof, to FcRn as compared with the FcRn binding activity of a parent albumin, reference albumin, fragment thereof or a fusion polypeptide comprising said parent albumin, reference albumin or fragment or fusion thereof, comprising contacting FcRn with said polypeptide, wherein said polypeptide further comprises a substitution to K, L, or M at a position corresponding to 550 in SEQ ID NO: 2, a substitution to P, Y, W, H, F, T, I, or V at a position corresponding to 573 in SEQ ID NO: 2, and a substitution to K or R at a position corresponding to 580 in SEQ ID NO: 2, wherein said polypeptide has a stronger binding affinity to FcRn or longer plasma half-life than a parent albumin, reference albumin, fragment thereof or fusion polypeptide comprising said parent albumin, reference albumin or fragment or fusion thereof.
2. The method of claim 1 , wherein within said polypeptide further comprises:
a) a substitution to A, C, D, E, F, G, H, I, L, M, N P, Q, R, S, T, V, W, or Y at a position corresponding to position 574 of SEQ ID NO: 2; or
b) a substitution to N at a position corresponding to position 574 of SEQ ID NO: 2, a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2, and a substitution at a position corresponding to position 492 of SEQ ID NO: 2.
3. The method of claim 1 , wherein within said polypeptide further comprises:
a substitution to P at a position corresponding to position 573 of SEQ ID NO: 2, a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2, and a substitution at a position corresponding to position 492 or 574 of SEQ ID NO: 2.
4. The method of claim 1 , wherein within said polypeptide further comprises:
a) a substitution to A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, or Y, at a position corresponding to position 492 of SEQ ID NO: 2; or
b) a substitution to N at a position corresponding to position 492 of SEQ ID NO: 2, a substitution to K at a position corresponding to position 573 of SEQ ID NO: 2, and a substitution at a position corresponding to position 574 of SEQ ID NO: 2.
5. The method of claim 1 , wherein said polypeptide further comprises a substitution at one or more positions selected from positions corresponding to positions 492 and 574 in SEQ ID NO: 2.
6. The method of claim 1 , wherein the reference albumin is HSA (SEQ ID NO: 2) or a fragment thereof, or a fusion polypeptide comprising HSA or a fragment thereof.
7. The method of claim 1 , wherein the sequence identity of said polypeptide to SEQ ID NO: 2 is more than 80%, more than 90%, more than 95%, more than 96%, more than 97%, more than 98% or more than 99%.
8. The method of claim 1 , wherein said fusion comprises a fusion partner polypeptide selected from a therapeutic, prophylactic, diagnostic, imaging or other beneficial moiety.
9. The method of claim 1 , wherein said polypeptide further comprises a non-albumin moiety covalently attached to said polypeptide.
10. The method of claim 1 , wherein said polypeptide further comprises a non-albumin moiety noncovalently associated with said polypeptide.
11. A method for altering the circulating half-life of a molecule comprising providing a variant of albumin, a fragment thereof or a fusion polypeptide comprising said variant albumin or fragment thereof; and
a) where the molecule is a polypeptide, fusing or conjugating the molecule to said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof; or
b) where the molecule is not a polypeptide, conjugating the molecule to said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof; or
c) contacting the molecule with said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof, wherein said contacting results in a noncovalent association between said molecule and variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof;
wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to K, L, or M at a position corresponding to 550 in SEQ ID NO: 2, a substitution to P, Y, W, H, F, T, I, or V at a position corresponding to 573 in SEQ ID NO: 2, and a substitution to K or R at a position corresponding to 580 in SEQ ID NO: 2, wherein said polypeptide has a stronger binding affinity to FcRn or longer plasma half-life than a parent albumin, reference albumin, fragment thereof or fusion polypeptide comprising said parent albumin, reference albumin or fragment or fusion thereof.
12. The method of claim 11 , wherein within said polypeptide further comprises:
a) a substitution to A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, or Y at a position corresponding to position 574 of SEQ ID NO: 2; or
b) a substitution to N at a position corresponding to position 574 of SEQ ID NO: 2, a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2, and a substitution at a position corresponding to position 492 of SEQ ID NO: 2.
13. The method of claim 11 , wherein within said polypeptide further comprises:
a substitution to P at a position corresponding to position 573 of SEQ ID NO: 2, a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2, and a substitution at a position corresponding to a position selected from the group consisting of 492 and 574 of SEQ ID NO: 2.
14. The method of claim 11 , wherein within said polypeptide further comprises:
a) a substitution to A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, or Y at a position corresponding to position 492 of SEQ ID NO: 2; or
b) a substitution to N at a position corresponding to position 492 of SEQ ID NO: 2, a substitution to K at a position corresponding to position 573 of SEQ ID NO: 2, and a substitution at a position corresponding to position 574 of SEQ ID NO: 2.
15. The method of claim 11 , wherein said polypeptide further comprises a substitution at one or more positions selected from positions corresponding to positions 492 and 574 in SEQ ID NO: 2.
16. The method of claim 1 , wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to P, Y, or W at a position corresponding to 573.
17. The method of claim 1 , wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to P at a position corresponding to 573.
18. The method of claim 1 , wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to L at a position corresponding to 550, a substitution to P at a position corresponding to 573, and a substitution to K at a position corresponding to 580.
19. The method of claim 11 , wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to P, Y, or W at a position corresponding to 573.
20. The method of claim 11 , wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to P at a position corresponding to 573.
21. The method of claim 11 , wherein said variant of albumin, fragment thereof or fusion polypeptide comprising said variant albumin or fragment thereof comprises a substitution to L at a position corresponding to 550, a substitution to P at a position corresponding to 573, and a substitution to K at a position corresponding to 580.
22. The method of claim 1 , wherein said variant of albumin, fragment thereof or fusion polypeptides comprising said variant albumin or fragment thereof comprises a substitution to M at a position corresponding to position 550 of SEQ ID NO: 2, a substitution to P at a position corresponding to position 573 of SEQ ID NO: 2, and a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2.
23. The method of claim 1 , wherein said variant of albumin, fragment thereof or fusion polypeptides comprising said variant albumin or fragment thereof comprises a substitution to K at a position corresponding to position 550 of SEQ ID NO: 2, a substitution to P at a position corresponding to position 573 of SEQ ID NO: 2, and a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2.
24. The method of claim 11 , wherein said variant of albumin, fragment thereof or fusion polypeptides comprising said variant albumin or fragment thereof comprises a substitution to M at a position corresponding to position 550 of SEQ ID NO: 2, a substitution to P at a position corresponding to position 573 of SEQ ID NO: 2, and a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2.
25. The method of claim 11 , wherein said variant of albumin, fragment thereof or fusion polypeptides comprising said variant albumin or fragment thereof comprises a substitution to K at a position corresponding to position 550 of SEQ ID NO: 2, a substitution to P at a position corresponding to position 573 of SEQ ID NO: 2, and a substitution to K at a position corresponding to position 580 of SEQ ID NO: 2.