Cell culture compositions with antioxidants and methods for polypeptide production

Patent metadata

Field	Value
Publication number	US-10017732-B2
Application number	US-201414211416-A
Country	US
Kind code	B2
Filing date	Mar 14, 2014
Priority date	Mar 15, 2013
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

Cell culture media comprising antioxidants are provided herein as are methods of using the media for cell culturing and polypeptide production from cells. Compositions comprising polypeptides, such as therapeutic polypeptides, produced by the methods herein are also provided.

First claim

The invention claimed is:

1. A method of producing a recombinant antibody composition with reduced color intensity, comprising the steps of:
culturing a cell comprising a nucleic acid encoding the recombinant antibody in a cell culture medium, wherein the cell culture medium comprises one or more of components (a)-(d):
(a) hypotaurine,
(b) s-carboxymethylcysteine,
(c) butylated hydroxyanisole, and
(d) quercitrin hydrate; and

producing the recombinant antibody;

wherein the cell culture medium comprising one or more of components (a)-(d) reduces the color intensity of a composition comprising the recombinant antibody produced by the cell as compared to a composition comprising the recombinant antibody produced by the cell cultured in a cell culture medium that does not comprise one or more of components (a)-(d).

2. The method of claim 1 , wherein the recombinant antibody is an antibody fragment.

3. The method of claim 1 , wherein the recombinant antibody is an IgG1 antibody.

4. The method of claim 1 , 2 or 3 , wherein the recombinant antibody is secreted into the cell culture medium.

5. The method of claim 1 , further comprising the step of recovering the recombinant antibody from the cell culture medium comprising one or more of components (a)-(d).

6. The method of claim 5 , wherein a composition comprising the recovered recombinant antibody is a liquid composition or a non-liquid composition.

7. The method of claim 6 , wherein the composition comprising the recovered recombinant antibody appears as a colorless or slightly colored liquid.

8. The method of claim 1 , wherein the cell culture medium comprising one or more of components (a)-(d) reduces the color intensity of a composition comprising the recombinant antibody produced by the cells by at least about 0.1% as compared to a composition comprising the recombinant antibody produced by the cell cultured in a cell culture medium that does not comprise the one or more of components (a)-(d).

9. The method of claim 1 , wherein the cell culture medium comprising one or more of components (a)-(d) reduces the color intensity of a composition comprising the recombinant antibody produced by the cells by about 5% to about 50% as compared to a composition comprising the recombinant antibody produced by the cell cultured in a cell culture medium that does not comprise the one or more of components (a)-(d).

10. The method of claim 1 , wherein the cell culture medium comprises the one or more components (a)-(d) in an amount selected from:
(a) hypotaurine at a concentration from at least about 0.0001 mM;
(b) s-carboxymethylcysteine at a concentration from at least about 0.0001 mM;
(c) butylated hydroxyanisole at a concentration from at least about 0.0001 mM; and
(d) quercitrin hydrate at a concentration from at least about 0.0001 mM.

11. The method of claim 10 , wherein the cell culture medium comprises hypotaurine at a concentration from about 2.0 mM to about 50.0 mM.

12. The method of claim 10 , wherein the cell culture medium comprises s-carboxymethylcysteine at a concentration from about 8.0 mM to about 12.0 mM.

13. The method of claim 10 , wherein the cell culture medium comprises butylated hydroxyanisole at a concentration from about 0.025 mM to about 0.040 mM.

14. The method of claim 10 , wherein the cell culture medium comprises quercitrin hydrate at a concentration from about 0.010 mM to about 0.020 mM.

15. The method of claim 1 , wherein the cell culture medium is a chemically defined cell culture medium.

16. The method of claim 1 , wherein the cell culture medium is a chemically undefined cell culture medium.

17. The method of claim 1 , wherein the cell culture medium is a basal cell culture medium.

18. The method of claim 1 , wherein the cell culture medium is a feed cell culture medium.

19. The method of claim 1 , wherein the cell is contacted with the cell culture medium during the cell's growth phase.

20. The method of claim 1 , wherein the cell is contacted with the cell culture medium during the cell's production phase.

21. The method of claim 1 , wherein the one or more of components (a)-(d) is added to the cell culture medium on at least one day of a cell culture cycle.

22. The method of claim 1 , wherein the one or more of components (a)-(d) is added to the cell culture medium on day 0 of a 14 day cell culture cycle.

23. The method of claim 1 , wherein the cell is a mammalian cell.

24. The method of claim 23 , wherein the mammalian cell is a Chinese Hamster Ovary (CHO) cell.

Assignees

• Genentech Inc

Inventors

• Vijayasankaran Natarajan
• Meier Steven J
• Varma Sharat
• Yang Yi

Classifications

• C12N5/0018 Primary

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12N2500/44

  Thiols, e.g. mercaptoethanol · CPC title

• C12N2500/30

  Organic components · CPC title

• C12N2500/33

  other than alpha-amino carboxylic acids, e.g. beta-amino acids, taurine · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title