Method for increasing the specific production rate of eukaryotic cells

• US 20170152473A1
• Filed: 02/09/2017
• Published: 06/01/2017
• Est. Priority Date: 08/11/2014
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Popp, Oliver
• Granted Patent: US 10,336,983 B2
• Time in Patent Office: Days
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/33 other than alpha-amino carboxylic acids, e.g. beta-amino acids, taurine
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2510/02 Cells for production
• CPC Class Codes: C12N 5/0018 Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0025 Culture media for plant cell or plant tissue culture
• CPC Class Codes: C12N 5/0682 Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells oocytes C12N5/0609

Abstract

The current invention reports the use of meta-tyrosine for increasing the specific productivity of a eukaryotic cell that produces/expresses a polypeptide. In the current method it is not necessary to perform a temperature-, osmolality- or pH shift or to add drugs like valproic acid or sodium butyrate to modulate the specific productivity of the cultivated cells. The method does not affect cell viability or product titer.

Claims

1. A method for increasing specific productivity (qP) of a eukaryotic host cell, the method comprising culturing the cell in culture medium, wherein the culture medium contains meta-tyrosine.

2. The method of claim 1, wherein the eukaryotic host cell is a mammalian cell.

3. The method of claim 2, wherein the mammalian cell is a Chinese Hampster Ovary (CHO) cell.

4. The method of claim 1, wherein the eukaryotic host cell produces an exogenous polypeptide.

5. The method of claim 1, wherein the eukaryotic host cell is cultured in suspension.

6. The method of claim 1, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.2 mM to 0.7 mM.

7. The method of claim 1, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.25 mM to 0.6 mM.

8. The method of claim 1, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.5 mM.

9. The method of claim 1, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.4 mM.

10. The method of claim 1, wherein the culture medium further comprises phenylalanine in a non-limiting concentration.

11. The method of claim 10, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 1.25.

12. The method of claim 10, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.25.

13. The method of claim 10, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.125.

14. The method of claim 10, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.025.

15. The method of claim 4, wherein the exogenous polypeptide is an immunoglobulin or a variant thereof or a fragment thereof or a fusion thereof.

16. A method for producing an exogenous polypeptide in a eukaryotic host cell, wherein the host cell expresses a nucleic acid encoding the exogenous polypeptide, the method comprising culturing the eukaryotic host cell in a culture medium comprising meta-tyrosine.

17. The method of claim 16, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.2 mM to 0.7 mM.

18. The method of claim 16, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.25 mM to 0.6 mM.

19. The method of claim 16, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.5 mM.

20. The method of claim 16, wherein the meta-tyrosine in the culture medium is at a concentration of from 0.3 mM to 0.4 mM.

21. The method of claim 16, wherein the eukaryotic host cell is a mammalian cell.

22. The method of claim 16, wherein the eukaryotic host cell is a Chinese Hampster Ovary (CHO) cell.

23. The method of claim 16, wherein the polypeptide is an immunoglobulin or a variant thereof or a fragment thereof or a fusion thereof.

24. The method of claim 16, wherein the culture medium additionally comprises phenylalanine in a non-limiting concentration.

25. The method of claim 24, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 1.25.

26. The method of claim 24, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.25.

27. The method of claim 24, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.125.

28. The method of claim 24, wherein the molar ratio of meta-tyrosine/phenylalanine is lower than or equal to 0.025.

29. The method of claim 16, wherein the culture medium temperature is kept constant during the process.

30. A eukaryotic cell culture medium comprising meta-tyrosine and phenylalanine in a molar ratio which is lower than or equal to 1.25.

31. The cell culture medium of claim 30, wherein the culture medium is a chemically defined culture medium.