| 主权项 |
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,YE492A,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; orG584A,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. |
