Cell culture medium

Patent metadata

Field	Value
Publication number	US-9428727-B2
Application number	US-201113643998-A
Country	US
Kind code	B2
Filing date	Apr 25, 2011
Priority date	Apr 26, 2010
Publication date	Aug 30, 2016
Grant date	Aug 30, 2016

Abstract

The present application describes an optimized medium for growth of mammalian cells as well as polypeptide production. The cell culture medium is characterized by a Sow ratio of sodium to potassium ions, it further relates to the method of producing polypeptides using such cell culture media. Sn another aspect, the method of polypeptide production can also comprise a temperature shift and/or a pH-shift to further optimize growth and product yield.

First claim

The invention claimed is:

1. A serum-free and protein-free cell culture medium for expressing recombinant polypeptides from Chinese Hamster Ovary, Human Embryonic Kidney or SP2/0 myeloma cells having a molar ratio of sodium to potassium ions of between 8 to 1 and 6 to 1, and wherein the sodium ions have a concentration of between 50 and 90 mM.

2. The cell culture medium of claim 1 wherein the potassium ions have a concentration of between 8 and 12 mM.

3. The cell culture medium of claim 1 comprising a total amino acid concentration between 40 and 100 mM.

4. The cell culture medium of claim 3 wherein the molar ratio between the total ion concentration to the total amino acid concentration is between 1.9 and 4.

5. A process for the production of a recombinant polypeptide comprising culturing mammalian cells comprising a recombinant nucleic acid encoding the recombinant polypeptide in the medium of claim 1 and expressing the recombinant polypeptide, wherein the mammalian cells are selected from the group consisting of Chinese Hamster Ovary (CHO) cells, Human Embryonic Kidney (HEK) cells, and SP2/0 cells.

6. The process according to claim 5 wherein culturing conditions comprise at least one temperature shift and/or at least one pH shift.

7. The process according to claim 5 , wherein culturing is done by a fed batch process.

8. The process according to claim 5 wherein the expressed recombinant polypeptide is glycosylated.

9. The process according to claim 5 , wherein the expressed polypeptide is an antibody or antibody fragment.

10. A process for the production of a cell culture medium according to claim 1 wherein the medium's components are admixed with each other.

11. The process according to claim 10 wherein sodium chloride is added at a concentration between 7 and 15 mM.

12. The process according to claim 10 wherein potassium chloride is added at a concentration between 8 and 12 mM.

Assignees

• Leist Christian
• Meissner Petra
• Schmidt Jörg
• Novartis Ag

Inventors

• Leist Christian
• Meissner Petra
• Schmidt Jörg

Classifications

• C12N5/005 Primary

  Protein-free medium · CPC title

• C12N2500/05

  Inorganic components · CPC title

• C12N5/0018 Primary

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

• C12N5/0031 Primary

  Serum-free culture media · CPC title

• C12N2500/12

  Light metals, i.e. alkali, alkaline earth, Be, Al, Mg · CPC title