Method of producing a polypeptide or virus of interest in a continuous cell culture

Patent metadata

Field	Value
Publication number	US-10745672-B2
Application number	US-201916252216-A
Country	US
Kind code	B2
Filing date	Jan 18, 2019
Priority date	Jul 31, 2009
Publication date	Aug 18, 2020
Grant date	Aug 18, 2020

Abstract

Described herein is a chemostat-like continuous cell culture system that combines certain advantages of perfusion open systems and chemostat open systems to improve the culturing of mammalian cells, e.g., genetically modified cells, particularly in serum-free or chemically-defined media. The continuous culture system described herein involves culturing mammalian cells in a continuous cell culture system, which comprises a cell retention device, wherein the cell culture system has a dilution rate (D) of less than about 2 d −1 , and a cell density of less than about 2×10 7 cell/mL. Also described herein is a method for producing a polypeptide and/or virus of interest in a continuous cell culture, the method comprising culturing mammalian cells expressing the polypeptide and/or virus of interest in a continuous cell culture system, which comprises a cell retention device, wherein the cell culture system has a dilution rate (D) of less than about 2 d −1 , and a cell density of less than about 2×10 7 cell/mL; and recovering the polypeptide and/or virus of interest from medium of the cell culture system.

First claim

The invention claimed is:

1. A method for producing a polypeptide of interest in a continuous cell culture, said method comprising
(a) culturing mammalian cells genetically modified to express the polypeptide of interest in a continuous cell culture system, wherein said cell culture system has a dilution rate (D) of less than 0.6 d −1 , a ratio of the dilution rate to a specific growth rate (D/μ) of from 1.2 to 5, and a cell density of less than 5×10 6 cell/mL; and
(b) recovering the polypeptide of interest from culture medium removed from the cell culture system.

2. The method of claim 1 , wherein the cells are CHO cells.

3. The method of claim 1 , wherein the polypeptide of interest is a disintegrin-like and metallopeptidase with thrombospondin type 1 motif 13 (ADAMTS13) protein.

4. The method of claim 1 , wherein the culture system further comprises a cell retention device that produces a cell retention rate of from 50% to 90%.

5. The method of claim 1 , wherein said cells are cultured in a serum-free medium.

6. The method of claim 2 , wherein the polypeptide of interest is a disintegrin-like and metallopeptidase with thrombospondin type 1 motif 13 (ADAMTS13) protein.

7. The method of claim 2 , wherein the culture system further comprises a cell retention device that produces a cell retention rate of from 50% to 90%.

8. The method of claim 2 , wherein said cells are cultured in a serum-free medium.

9. The method of claim 3 , wherein the culture system further comprises a cell retention device that produces a cell retention rate of from 50% to 90%.

10. The method of claim 3 , wherein said cells are cultured in a serum-free medium.

11. The method of claim 4 , wherein said cells are cultured in a serum-free medium.

12. The method of claim 6 , wherein the culture system further comprises a cell retention device that produces a cell retention rate of from 50% to 90%.

13. The method of claim 6 , wherein said cells are cultured in a serum-free medium.

14. The method of claim 7 , wherein said cells are cultured in a serum-free medium.

15. The method of claim 9 , wherein said cells are cultured in a serum-free medium.

16. The method of claim 12 , wherein said cells are cultured in a serum-free medium.

17. The method of claim 1 , wherein said cell culture system has a specific growth rate of between 0.15 d −1 and 0.3 d −1 .

18. The method of claim 1 , wherein said cells are cultured in said cell culture system for more than 40 days.

19. The method of claim 4 , wherein said cell retention device comprises a macroporous microcarrier.

20. The method of claim 1 , wherein said cells are cultured in at least 250 L of medium.

Assignees

• Baxalta GmbH
• Baxalta Inc

Inventors

• Grillberger Leopold
• Reiter Manfred
• Fleischanderl Daniel

Classifications

• C12Y304/24087

  ADAMTS13 endopeptidase (3.4.24.87) · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title

• C12N2500/98

  Xeno-free medium and culture conditions · CPC title

• C12N7/00 Primary

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N2710/00052

  relating to complementing cells and packaging systems for producing virus or viral particles · CPC title