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Last Updated: March 28, 2024

Claims for Patent: 7,829,309


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Summary for Patent: 7,829,309
Title:Cell culture performance with betaine
Abstract: This invention relates generally to the field of cell culture. More particularly, the invention relates to improving viability of recombinant cell cultures and the yields of secreted polypeptides therefrom by the addition of betaine to the tissue culture medium.
Inventor(s): Follstad; Brian D. (Seattle, WA), Potter; Anne H. (Sammamish, WA)
Assignee: Immunex Corporation (Thousand Oaks, CA)
Application Number:12/119,206
Patent Claims:1. A cell culture comprising a recombinantly engineered cell line in tissue culture medium, wherein the cell line is recombinantly engineered to express a protein of interest, the medium has a concentration of betaine from about 1 mM to about 100 mM in an amount effective to improve cell survival and expression of said protein of interest and the tissue culture medium is not hyperosmotic.

2. The cell culture of claim 1, wherein the recombinantly engineered cell line is a non-animal cell line.

3. The cell culture of claim 2, wherein the cell line is a prokaryotic cell line.

4. The cell culture of claim 3, wherein the cell line is an E. coli cell line.

5. The cell culture of claim 2, wherein the cell line is an insect cell line.

6. The cell culture of claim 5, wherein the cell line is an Sf9 cell line.

7. The cell culture of claim 2, wherein the cell line is a plant cell line.

8. The cell culture of claim 2, wherein the cell line is a yeast cell line.

9. The cell culture of claim 1, wherein the betaine is selected from the group consisting of glycine betaine and betaine aldehyde.

10. The cell culture of claim 1, wherein the betaine is at a concentration of 5 mM to 75 mM.

11. The cell culture of claim 1, wherein the betaine is at a concentration of 10 mM to 60 mM.

12. The cell culture of claim 1, wherein the betaine is at a concentration of about 20 mM to 40 mM.

13. The cell culture of claim 1, wherein the betaine is at a concentration selected from the group consisting of 10 mM, 20 mM, 30 mM, and 40 mM.

14. The cell culture of claim 1, wherein the betaine is at a concentration of about 20 mM.

15. The cell culture of claim 1, wherein the protein of interest is selected from the group consisting of a soluble TNF receptor, a soluble IL-4 receptor, a soluble IL-1 type II receptor, a soluble flt3 ligand, a soluble CD40 ligand, an erythropoietin, an antibody, an Fc-fusion protein, a calcitonin, a growth hormone, an insulin, an insulinotropin, insulin-like growth factors, a parathyroid hormone, an interferons, a nerve growth factor, a glucagons, an interleukins, a colony stimulation factor, a glucocerebrosidase, a superoxide dismutase, a tissue plasminogen activator, a Factor VIII, a Factor IX, an aploipoprotein E, an Apolipoprotein A-I, a globin, an IL-2 receptor, an IL-2 antagonist, alpha-1 antitrypsin, and an alpha-galactosidase A.

16. The cell culture of claim 15, wherein the cell culture is in a bioreactor.

17. The cell culture of claim 1, wherein the cell culture is in suspension culture.

18. The cell culture of claim 15 wherein the cell line is an animal cell line.

19. The cell culture of claim 18 wherein the animal cell line is a Chinese hamster ovary (CHO) cell line.

20. The cell culture of claim 18 or 19 wherein the protein of interest is an antibody.

21. The cell culture of claim 20 wherein the antibody is a human antibody.

22. The cell culture of claim 20 wherein the antibody is a humanized antibody.

23. The cell culture of claim 19, wherein the protein of interest is selected from the group consisting of a soluble TNF receptor Fc fusion, an anti-EGF Receptor antibody, an anti-TNF alpha antibody, an anti-CD20 antibody, and an anti-VEGF antibody.

24. The cell culture of claim 23, wherein the medium is serum-free medium.

25. An animal cell culture comprising a recombinantly engineered cell line in serum-free culture medium, wherein the cell line is recombinantly engineered to express an antibody to a human EGF receptor, the medium has a concentration of betaine from about 1 mM to about 100 mM, and the tissue culture medium is not hyperosmotic.

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