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Generated: December 17, 2017

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Title: Human growth hormone variants comprising amino acid substitutions
Abstract:Human growth hormone variants are disclosed having enhanced affinity for the growth hormone receptor. Also disclosed are human growth hormone variants conjugated to one or more chemical groups, such as poly(ethylene glycol), which is believed to prolong the in vivo half-life of the variants.
Inventor(s): Cunningham; Brian C. (San Bruno, CA), Wells; James A. (Burlingame, CA), Clark; Ross G. (Pacifica, CA), Olson; Kenneth (Burlingame, CA), Fuh; Germaine G. (Pacifica, CA)
Assignee: Genentech, Inc. (San Francisco, CA)
Filing Date:Feb 24, 1998
Application Number:09/028,652
Claims:1. A variant of human growth hormone wherein the variant comprises all of the following amino acid substitutions: H18A, Q22A, F25A, D26A, Q29A, E65A, K168A, E174A, and further comprising a substitution at G120.

2. The human growth hormone variant of claim 1, wherein the substitution at G120 is selected from the group consisting of G120R, G120K, G120W, G120Y, G120F, and G120E.

3. The human growth hormone variant of claim 2 wherein the substitution at G120 is G120K.

4. The human growth hormone variant of claim 3, 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.

5. The human growth hormone variant of claim 4, wherein said chemical group is poly(ethylene glycol).

6. The human growth hormone variant of claim 5 where the human growth hormone variant is conjugated to between about four and about six molecules of poly(ethylene glycol).

7. 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.

8. The human growth hormone variant of claim 7, wherein said chemical group is poly(ethylene glycol).

9. A composition comprising the human growth hormone variant of claim 8, wherein said human growth hormone variant is conjugated to between about four and about six molecules of poly(ethylene glycol).

10. 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.

11. A nucleic acid encoding a human growth hormone variant comprising all of the following amino acid substitutions: H18A, Q22A, F25A, D26A, Q29A, E65A, K168A, E174A, and further comprising a substitution at G120K.

12. A vector comprising the nucleic acid of claim 11.

13. A host cell comprising the vector of claim 12.

14. A process for preparing a human growth hormone variant comprising culturing the host cell of claim 13 and recovering the human growth hormone variant from the culture.

15. A nucleic acid encoding a human growth hormone variant comprising all of the following amino acid substitutions: H18A, Q22A, F25A, D26A, Q29A, E65A, K168A, E174A and G120K.

16. A vector comprising the nucleic acid sequence of claim 15.

17. A host cell comprising the vector of claim 16.
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