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Details for Patent: 5,688,666

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Details for Patent: 5,688,666

Title: Growth hormone variants with altered binding properties
Abstract:A method for selecting novel proteins such as growth hormone variants having altered binding properties for a growth hormone receptor molecule is provided. The method comprises fusing a gene encoding a hormone to the carboxy terminal domain of the gene III coat protein of the filamentous phage M13. The gene fusion is mutated to form a library of structurally related fusion proteins that are expressed in low quantity on the surface of a phagemid particle. Biological selection and screening are employed to identify novel ligands useful as drug candidates. Disposed are preferred phagemid expression vectors and selected human growth hormone variants.
Inventor(s): Bass; Steven H. (Redwood City, CA), Greene; Ronald (Durham, NC), Lowman; Henry B. (Hercules, CA), Wells; James A. (Burlingame, CA)
Assignee: Genentech, Inc. (San Francisco, CA)
Filing Date:Jan 14, 1994
Application Number:08/182,530
Claims:1. A variant of a native human growth hormone, said variant capable of binding to human growth hormone binding protein and having a set of amino acid substitutions selected from the group consisting of:

F10H,M14G,H18N,H21N;

F10A,M14W,H18D,H21N;

F10Y,M14T,H18V,H21N;

F10I,M14N,H18I,H21N; and

M14S,H18F,H21L, numbered from the N-terminus of 191-amino acid human growth hormone.

2. The variant of claim 1 wherein the set of amino acid substitutions is F10H,M14G,H18N,H21N.

3. The variant of claim 1 wherein the set of amino acid substitutions is F10A,M14W,H18D,H21N.

4. The variant of claim 1 wherein the set of amino acid substitutions is F10Y,M14T,H18V,H21N.

5. The variant of claim 1 wherein the set of amino acid substitutions is F10I,M14N,H18I,H21N.

6. The variant of claim 1 wherein the set of amino acid substitutions is M14S,H18F,H21L.

7. A variant of a native human growth hormone, said variant capable of binding to human growth hormone binding protein and comprising a set of amino acid substitutions selected from the group consisting of:

F10H,M14G,H18N,H21N;

F10A,M14W,H18D,H21N;

F10Y,M14T,H18V,H21N;

F10I,M14N,H18I,H21N; and

M14S,H18F,H21L, numbered from the N-terminus of 191-amino acid human growth hormone.

8. The variant of claim 7 wherein the affinity of said variant for human growth hormone binding protein is greater than that of said native human growth hormone and said set of amino acid substitutions is selected from the group consisting of:

F10H,M14G,H18N,H21N; and

F10A,M14W,H18D,H21N, numbered from the N-terminus of 191-amino acid human growth hormone.

9. The variant of claim 7 wherein the set of amino acid substitutions is F10H,M14G,H18N,H21N.

10. The variant of claim 7 wherein the set of amino acid substitutions is F10A,M14W,H18D,H21N.

11. The variant of claim 7 wherein the set of amino acid substitutions is F10Y,M14T,H18V,H21N.

12. The variant of claim 7 wherein the set of amino acid substitutions is F10I,M14N,H18I,H21N.

13. The variant of claim 7 wherein the set of amino acid substitutions is M14S,H18F,H21L.

14. A variant of a native human growth hormone, said variant capable of binding to human growth hormone binding protein and having a set of amino acid substitutions selected from the group consisting of:

R167N,D171S,E174S,F176Y,I173T;

R167E,D171S,E174S,F176Y;

R167K,D171S,E174S,F176Y,I179L;

R167N,D171N,E174S,F176Y,I179T;

R167N,D171S,E174S,F176Y,I179O; and

E174S,F176Y, numbered from the N-terminus of 191-amino acid human growth hormone.

15. The variant of claim 14 wherein the set of amino acid substitutions is R167N,D171S,E174S,F176Y,I179T.

16. The variant of claim 14 wherein the set of amino acid substitutions is R167E,D171S,E174S,F176Y.

17. The variant of claim 14 wherein the set of amino acid substitutions is R167K,D171S,E174S,F176Y,I179L.

18. The variant of claim 14 wherein the set of amino acid substitutions is R167N, D171N,E174S,F176Y,I179T.

19. The variant of claim 14 wherein the set of amino acid substitutions is R167N,D171S,E174S,F176Y,I179Q.

20. The variant of claim 14 wherein the set of amino acid substitutions is E174S,F176Y.

21. A variant of a native human growth hormone, said variant capable of binding to human growth hormone binding protein and comprising a set of amino acid substitutions selected from the group consisting of:

R167N,D171S,E174S,F176Y,I179T;

R167E,D171S,E174S,F176Y;

R167K,D171S,E174S,F176Y,I179L;

R167N,D171N,E174S,F176Y,I179T;

R167N,D171S,E174S,F176Y,I179Q; and

E174S,F176Y, numbered from the N-terminus of 191-amino acid human growth hormone.

22. The variant of claim 21 wherein the set of amino acid substitutions is R167N,D171S,E174S,F176Y,I179T.

23. The variant of claim 21 wherein the set of amino acid substitutions is R167E,D171S,E174S,F176Y.

24. The variant of claim 21 wherein the set of amino acid substitutions is R167K,D171S,E174S,F176Y,I179L.

25. The variant of claim 21 wherein the set of amino acid substitutions is R167N,D171N,E174S,F176Y,I179T.

26. The variant of claim 21 wherein the set of amino acid substitutions is R167N,D171S,E174S,F176Y,I179Q.

27. The variant of claim 21 wherein the set of amino acid substitutions is E174S,F176Y.

28. The variant of claim 21 wherein the affinity of said variant for human growth hormone binding protein is greater than that of said native human growth hormone.
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