Oligopeptide-free cell culture media

Patent metadata

Field	Value
Publication number	US-10696731-B2
Application number	US-201715670217-A
Country	US
Kind code	B2
Filing date	Aug 7, 2017
Priority date	Jan 4, 2006
Publication date	Jun 30, 2020
Grant date	Jun 30, 2020

Abstract

The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg/L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg/L of a polyamine.

First claim

What is claimed is:

1. A method for increasing the expression of a recombinant protein in a cell culture, the method comprising culturing mammalian cells that express a recombinant protein in a protein-free medium that does not comprise oligopeptides, wherein the medium comprises DMEM:HAM's F12 (1:1) basal medium, inorganic salts, amino acids, vitamins, ornithine, and putrescine at a concentration of at least 1 mg/L, and wherein the medium is supplemented with Fe(II) and Cu(II) at an amount greater than the amount in basal DMEM: HAM's F12 (1:1).

2. The method according to claim 1 , wherein the medium is supplemented with L-glutamine, ascorbic acid, ethanolamine, sodium selenite, and a non-ionic surfactant.

3. The method according to claim 1 , wherein the medium is supplemented with L-Asparagine, L-Cysteine, L-Cystine, L-Proline, and L-Tryptophan.

4. The method according to claim 1 , wherein the medium comprises putrescine at a concentration of from about 1.0 mg/L to about 20 mg/L.

5. The method according to claim 1 , wherein the medium further comprises one or more from the group consisting of cadaverine, spermidine, spermine, and agmatine.

6. The method according to claim 1 , wherein the medium further comprises spermine.

7. The method according to claim 1 , wherein the medium comprises a buffer substance.

8. The method according to claim 7 , wherein the buffer substance is selected from one or more of sodium bicarbonate, antioxidants, stabilizers, or protease inhibitors.

9. The method according to claim 1 , wherein the recombinant protein is selected from the group of coagulation factor VII, coagulation factor VIII, coagulation factor IX, vWF, ADAMTS13, and furin.

10. The method according to claim 8 , wherein the recombinant protein is coagulation factor VIII.

11. The method according to claim 8 , wherein the recombinant protein is vWF.

12. The method according to claim 1 , wherein the mammalian cells are CHO cells, 293 cells or BHK cells.

13. The method according to claim 1 , wherein the mammalian cell/protein combination is selected from the group consisting of CHO cells/coagulation factor VIII, CHO cells/coagulation factor VII, CHO cells/ADAMTS13, CHO cells/furin, 293 cells/coagulation factor IX.

14. The method according to claim 13 , wherein the mammalian cell/protein combination is CHO cells/coagulation factor VIII.

15. The method according to claim 1 , wherein the putrescine is synthetically produced.

16. The method according to claim 1 , wherein the putrescine originates from a source other than a protein hydrolysate.

17. The method according to claim 1 , wherein the medium is chemically defined.

18. The method according to claim 1 , wherein the mammalian cells are cultured by a method selected from the group of batch-cultivation, feed-batch-cultivation, perfusion-cultivation and chemostat-cultivation.

Assignees

• Baxalta GmbH
• Baxalta Inc

Inventors

• Grillberger Leopold
• Reiter Manfred
• Mundt Wolfgang
• Mitterer Artur

Classifications

• C12N5/005 Primary

  Protein-free medium · CPC title

• C12N2710/24151

  Methods of production or purification of viral material · CPC title

• C12N2500/10

  Metals; Metal chelators · CPC title

• C12N2500/32

  Amino acids · CPC title

• C12N2500/24

  Iron; Fe chelators; Transferrin · CPC title