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Claims for Patent: 5,849,535

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Claims for Patent: 5,849,535

Title: Human growth hormone variants
Abstract:Human growth hormone variants, DNA encoding the variants, vectors, host cells, pegylated forms of the variants, as well as methods of making the variants are disclosed.
Inventor(s): Cunningham; Brian C. (San Bruno, CA), Lowman; Henry B. (Hercules, CA), Wells; James A. (Burlingame, CA), Clark; Ross G. (Pacifica, CA), Olson; Kenneth (Burlingame, CA), Fuh; Germaine G. (Pacifica, CA)
Assignee: Genentech, Inc. (South San Francisco, CA)
Application Number:08/717,394
Patent Claims: 1. A human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T.

2. A method of producing a pegylated human growth hormone variant, comprising:

(a) pegylating the human growth hormone variant of claim 1;

(b) applying the pegylated human growth hormone variant to a cation exchange chromatography column; and

(c) eluting the pegylated human growth hormone variant.

3. The human growth hormone variant of claim 1, wherein the human growth hormone variant is conjugated to one or more chemical groups that increase the actual molecular weight of the human growth hormone variant to between about 40 and about 100 kilodaltons.

4. The human growth hormone variant of claim 3, wherein said chemical group is polyethylene glycol.

5. A composition comprising the human growth hormone variant of claim 4 wherein the human growth hormone variant is conjugated to between about four to about six molecules of polyethylene glycol.

6. The human growth hormone variant of claim 1, further comprising a substitution at G120.

7. The human growth hormone variant of claim 6, wherein the substitution at G120 is selected from the group consisting of G120R, G120K, G120W, G120Y, G120F, and G120E.

8. The human growth hormone variant of claim 7 wherein the substitution at G120 is G120K.

9. The human growth hormone variant of claim 8 wherein the human growth hormone variant is conjugated to one or more chemical groups that increase the actual molecular weight of the human growth hormone variant to between about 40 to about 100 kilodaltons.

10. The human growth hormone variant of claim 9 wherein said chemical group is polyethylene glycol.

11. The human growth hormone variant of claim 10 wherein the human growth hormone variant is conjugated to between about four to about six molecules of polyethylene glycol.

12. A nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T.

13. A nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T, and further comprising a substitution at G120.

14. A vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T.

15. A vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T, and further comprising a substitution at G120.

16. A host cell comprising a vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T.

17. A host cell comprising a vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T, and further comprising a substitution at G120.

18. A process for preparing a human growth hormone variant comprising culturing a host cell comprising a vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T and recovering the human growth hormone variant from the culture.

19. A process for preparing a human growth hormone variant comprising culturing a host cell comprising a vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, R167N, K168A, D171S, K172R, E174S, I179T, and further comprising a substitution at G120 and recovering the human growth hormone variant from the culture.

20. A nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, G120K, R167N, K168A, D171S, K172R, E174S, I179T.

21. A vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions: H18D, H21N, G120K, R167N, K168A, D171S, K172R, E174S, I179T.

22. A host cell comprising a vector comprising a nucleic acid encoding a human growth hormone variant comprising the following set of amino acid substitutions:

H18D, H21N, G120K, R167N, K168A, D171S, K172R, E174S, I179T.
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