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Last Updated: March 28, 2024

Claims for Patent: 4,775,622


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Summary for Patent: 4,775,622
Title: Expression, processing and secretion of heterologous protein by yeast
Abstract:Yeast organisms are caused to express, process, and secrete protein that is normally heterologous to yeast and not required for its viability by transforming yeast with an expression vector containing a gene encoding heterologous protein and a signal sequence also heterologous to yeast.
Inventor(s): Hitzeman; Ronald A. (Pacifica, CA), Leung; David W. (So. San Francisco, CA)
Assignee: Genentech, Inc. (South San Francisco, CA)
Application Number:06/438,236
Patent Claims:1. A yeast organism cell culture capable of expressing, processing and secreting a protein heterologous to said yeast organism comprising (1) viable yeast cells transformed with an expression vehicle functionally containing DNA encoding said protein together with a heterologous signal polypeptide therefor, said heterologous signal not being normally produced or employed by said yeast organism, and (2) a medium supporting said cell culture, said medium containing said protein as a product of said yeast organism expression, processing and secretion.

2. A cell culture according to claim 1 wherein the yeast organism is Saccharomyces cerevisiae.

3. A yeast organism cell culture capable of expressing, processing and secreting a protein heterologous to said yeast organism comprising (1) viable yeast cells transformed with an expression vehicle functionally containing DNA encoding said protein together with a heterologous signal polypeptide therefor, wherein said heterologous signal polypeptide is a hybrid of a signal native to said protein and of a second heterologous signal polypeptide and (2) a medium supporting said cell culture, said medium containing said protein as a product of said yeast organism expression, processing and secretion.

4. A yeast expression vehicle functionally containing DNA encoding a protein heterologous to a yeast organism and a heterologous signal polypeptide in translationl reading frame with the DNA encoding the heterologous protein wherein said heterologous signal polypeptide is a hybrid of a signal native to said protein and of a second heterologous signal polypeptide.

5. A process for obtaining protein heterologous to a yeast organism as a product of yeast expression, processing and secretion comprising (a) providing an expression vehicle functionally containing DNA encoding said protein and a heterologous signal peptide, said heterologous signal not being normally produced or employed by said yeast organism, (b) transforming with the expression vehicle a yeast organism capable of expressing and processing the protein and heterologous signal peptide and secreting the heterologous protein, (c) preparing a culture of said organism, (d) growing the culture and (e) recovering said protein from the medium of said culture.

6. The process of claim 5 wherein the recovered heterologous protein is separated from forms of the protein having additional amino acids of the heterologous signal polypeptide.

7. The process of claim 6 wherein the heterologous protein is human growth hormone.

8. The process of claim 5 wherein the heterologous protein is recovered from the culture medium when the culture has reached stationary phase.

9. The process of claim 5 wherein the vehicle contains tandem copies of DNA encoding the heterologous protein.

10. The process of claim 5 wherein the heterologous protein is a human protein.

11. The process of claim 5 wherein the heterologous protein is human serum albumin.

12. The process of claim 5 wherein the yeast organism is Saccharomyces cerevisiae and the protein is human growth hormone.

13. The process of claim 5 wherien the protein is a glycoprotein.

14. The process of claim 5 wherein the protein is a hormone, viral antigen, human serum albumin, human insulin or a glycoprotein.

15. A process for obtaining protein heterologous to a yeast organism as a product of yeast expression, processing and secretion comprising providing an expression vehicle functionally containing DNA encoding said protein and a heterologous signal peptide native to the heterologous protein, transforming with the expression vehicle a yeast organism capable of expressing and processing the protein and heterologous signal peptide and secreting the heterologous protein, preparing a culture of said organism, growing the culture, and recovering said protein from the medium of said culture.

16. A process for obtaining protein heterologous to a yeast organism as a product of yeast expression, processing and secretion comprising providing an expression vehicle functionally containing DNA encoding said protein and a heterologous signal peptide which is not from a baterial source, said heterologous signal not being normally produced or employed by said yeast organism, transforming with the expression vehicle a yeast organism capable of expressing and processing the protein and heterologous signal peptide and secreting the heterologous protein, preparing a culture of said organism, growing the culture and recovering said protein from the medium of said culture.

17. A yeast organism culture comprising (a) a yeast organism transformed with a yeast expression vehicle functionally containing DNA encoding a protein heterologous to the yeast organism and DNA encoding a heterologous signal polypeptide, said heterologous signal not being normally produced or employed by said yeast organism, and (b) culture medium containing said heterologous protein in mature form as a product of yeast secretion of the mature heterologous protein produced upon growth of the yeast organism in culture medium.

18. A yeast expression vehicle replicable in E. coli or in Saccharomyces cerevisiae, which vehicle contains DNA encoding a heterologous signal polypeptide in translational reading frame with DNA encoding said heterologous protein, wherein said signal polypeptide is a hybrid of a signal native to said protein and of a second heterologous signal polypeptide wherein said heterologous protein is expressed and secreted from a Saccharomyces cerevisiae host cell transformed with said vehicle.

19. A process for obtaining mature human growth hormone comprising transforming a yeast organism with an expression vehicle functionally containing DNA encoding pre human growth hormon wherein said yeast organism is capable of expressing and processing pre human growth hormone and secreting mature human growth hormone, preparing a culture of said yeast organism, growing the culture and recovering mature human growth hormone from the medium.

Details for Patent 4,775,622

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Grifols Therapeutics Llc ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5 albumin (human) For Injection 101138 10/21/1942 ⤷  Try a Trial 2005-10-04
Baxalta Us Inc. BUMINATE, FLEXBUMIN albumin (human) Injection 101452 03/03/1954 ⤷  Try a Trial 2005-10-04
Csl Behring Ag ALBURX albumin (human) Injection 102366 07/23/1976 ⤷  Try a Trial 2005-10-04
Grifols Biologicals Llc ALBUTEIN albumin (human) Injection 102478 08/15/1978 ⤷  Try a Trial 2005-10-04
Instituto Grifols, S.a. HUMAN ALBUMIN GRIFOLS albumin (human) Injection 103352 02/17/1995 ⤷  Try a Trial 2005-10-04
Instituto Grifols, S.a. HUMAN ALBUMIN GRIFOLS albumin (human) Injection 103352 06/11/2003 ⤷  Try a Trial 2005-10-04
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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