Albumin variants

Abstract

The present invention relates to variants of a parent albumin having altered plasma half-life compared with the parent albumin. The present invention also relates to fusion polypeptides and conjugates comprising said variant albumin.

1 . A Method for preparing a variant of albumin, fragments thereof or fusion polypeptide comprising said variant albumin or fragment thereof, comprising following steps:
 a. Providing a nucleic acid encoding a parent albumin having at least 80% sequence identity to SEQ ID NO: 2;   b. Modifying the sequence of step a., to encode a variant albumin, fragments thereof or fusion polypeptide comprising said variant albumin or fragment thereof having one or more substitutions corresponding to the substitutions in SEQ ID NO: 2 selected among:   Q417A,C,D,E,F,G,H,I,K,L,M,N,P, R,S,T,V,W,Y;   H440A,C,D,E,F,G, I,K,L,M,N,P,Q,R,S,T,V,W,Y;   A490C,D,E, F,G,H,I,K,L,M,N,P, R,S,T,V,W,Y   E492A,C,D,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   V493A,C,D,E,F,G,H,I,K,L,M,N,P,Q,R,S,T.W,Y;   D494A,C,E,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   E495A,C,D,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   T496A,C,D,E,F,G,H,I,K,L,M,N,P,Q,R,S,V,W,Y;   P499A,C,D,E,F,G,H,I,K,L,M,N, Q,R,S,T,V,W,Y;   K500A,C,D,E,F,G,H,I,L,M,N, Q,R,S,T,V,W,Y;   E501A,C,D,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   N503A,C,D,E,F,G,H,I,K,L,M, P,Q,R,S,T,V,W,Y;   A504C,D,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   E505A,C,D,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   T506A,C,D,F,G,H,I,K,L,M,N,P,Q,R,S, V,W,Y;   H510A,C,D,F,G, I,K,L,M,N,P,Q,R,S,T,V,W,Y;   H535A,C,D,F,G, I,K,L,M,N,P,Q,R,S,T,V,W,Y;   K536A,C,D,E,F,G,H,I, L,M,N,P,Q,R,S,T,V,W,Y;   P537A,C,D,E,F,G,H,I,K,L,M,N, Q,R,S,T,V,W,Y;   K538A,C,D,E,F,G,H,I, L,M,N,P,Q,R,S,T,V,W,Y;   T540A,C,D,E,F,G,H,I, L,M,N,P,Q,R,S,V,W,Y;   K541A,C,D,E,F,G,H,I, L,M,N,P,Q,R,S,T,V,W,Y;   E542A,C,D,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   D550A,C,E,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   K573A,C,D,E,F,G,H,I, L,M,N,P,Q,R,S,T,V,W,Y;   K574A,C,D,E,F,G,H,I, L,M,N,P,Q,R,S,T,V,W,Y;   Q580A,C,D,E,F,G,H,I, K, L,M,N,P,R,S,T,V,W,Y;   A581C,D,E,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   A582C,D,E,F,G,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y; or   G584A,C,D,E,F,H,I,K,L,M,N,P,Q,R,S,T,V,W,Y;   c. Introducing the modified sequence of step b., in a suitable host cell;   d. Growing the cells in a suitable growth medium under condition leading to expression of the variant of albumin, fragments thereof or fusion polypeptide comprising said variant albumin or fragment thereof; and   e. Recovering the variant of albumin, fragments thereof or fusion polypeptide comprising said variant albumin or fragment thereof from the growth medium;   wherein the variant of albumin, fragments thereof or fusion polypeptide comprising said variant albumin or fragment thereof, has an altered plasma half-life compared with the parent albumin, fragments thereof or fusion polypeptide comprising said variant albumin or fragment thereof.   
     
     
         2 . A fusion polypeptide comprising:
 a variant of albumin comprising a substitution in a position corresponding to position 573 in the amino acid sequence of SEQ ID NO:2, wherein the substitution is selected from the group consisting of K573A, C, D, F, G, H, I, L, M, N, P, Q, R, S, V, W, and Y, and wherein the variant has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2, and   a polypeptide fusion partner,   in which the variant of albumin is genetically fused to the polypeptide fusion partner.   
     
     
         3 . The fusion polypeptide of  claim 2 , wherein the polypeptide fusion partner comprises a therapeutically beneficial moiety or a diagnostically useful moiety. 
     
     
         4 . The variant of  claim 2 , wherein the variant has at least 99% sequence identity to the amino acid sequence of SEQ ID NO:2. 
     
     
         5 . The variant of  claim 2 , wherein the variant consists of the amino acid sequence of SEQ ID NO:2 with a substitution at position 573. 
     
     
         6 . The variant of  claim 2 , wherein the substitution is K573A. 
     
     
         7 . The variant of  claim 2 , wherein the substitution is K573C. 
     
     
         8 . The variant of  claim 2 , wherein the substitution is K573D. 
     
     
         9 . The variant of  claim 2 , wherein the substitution is K573F. 
     
     
         10 . The variant of  claim 2 , wherein the substitution is K573G. 
     
     
         11 . The variant of  claim 2 , wherein the substitution is K573H. 
     
     
         12 . The variant of  claim 2 , wherein the substitution is K573I. 
     
     
         13 . The variant of  claim 2 , wherein the substitution is K573L. 
     
     
         14 . The variant of  claim 2 , wherein the substitution is K573M. 
     
     
         15 . The variant of  claim 2 , wherein the substitution is K573N. 
     
     
         16 . The variant of  claim 2 , wherein the substitution is K573P. 
     
     
         17 . The variant of  claim 2 , wherein the substitution is K573Q. 
     
     
         18 . The variant of  claim 2 , wherein the substitution is K573R. 
     
     
         19 . The variant of  claim 2 , wherein the substitution is K573S. 
     
     
         20 . The variant of  claim 2 , wherein the substitution is K573V. 
     
     
         21 . The variant of  claim 2 , wherein the substitution is K573W. 
     
     
         22 . The variant of  claim 2 , wherein the substitution is K573Y. 
     
     
         23 . A conjugate comprising:
 a variant of albumin comprising a substitution in a position corresponding to position 573 in the amino acid sequence of SEQ ID NO:2, wherein the substitution is selected from the group consisting of K573A, C, D, F, G, H, I, L, M, N, P, Q, R, S, V, W, and Y, and wherein the variant has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2, and   a second molecule,   in which the variant of albumin is conjugated to the second molecule.   
     
     
         24 . The conjugate according to  claim 23 , wherein the second molecule comprises a therapeutically beneficial moiety or a diagnostically useful moiety.