Oligopeptide-free cell culture media

Patent metadata

Field	Value
Publication number	US-9758568-B2
Application number	US-201514942771-A
Country	US
Kind code	B2
Filing date	Nov 16, 2015
Priority date	Jan 4, 2006
Publication date	Sep 12, 2017
Grant date	Sep 12, 2017

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:

1. A method for expressing a coagulation factor VIII protein, comprising the steps of:
(a) providing a culture of CHO cells;
(b) introducing at least one nucleic acid sequence comprising a sequence coding for the coagulation factor VIII protein into the cells;
(c) selecting the cells carrying the nucleic acid sequence; and
(d) expressing the coagulation factor VIII protein in a chemically defined protein-free medium that does not comprise oligopeptides, the medium comprising DMEM:HAM's F12 (1:1) basal medium, putrescine at a concentration of at least 1 mg/L, and Fe(II) and Cu(II) at a level greater than the amount in basal DMEM:HAM's F12 (1:1), wherein the medium is supplemented with L-glutamine, ascorbic acid, ethanolamine, sodium selenite, and a non-ionic surfactant, wherein the cells are cultivated by chemostat cultivation.

2. The method of claim 1 , wherein the medium further comprises a polyamine selected from the group consisting of cadaverine, spermidine, spermine, agmatine, ornithine, or combinations thereof.

3. The method of claim 1 , wherein the medium comprises ornithine, spermine, or combinations thereof.

4. The method of claim 1 , wherein the putrescine is synthetically produced.

5. The method of claim 1 , wherein the putrescine originates from a source other than a protein hydrolysate.

6. The method of claim 1 , wherein the medium is further supplemented with L-Asparagine, L-Cysteine, L-Cystine, L-Proline, and L-Tryptophan.

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

8. The method of claim 1 , wherein the non-ionic surfactant comprises copolymers or mixtures of polyethylene glycols and polypropylene glycols.

9. The method of claim 1 , wherein the medium further comprises a buffer substance.

10. The method of claim 9 , wherein the buffer substance comprises one or more of sodium bicarbonate, antioxidants, stabilizers, or protease inhibitors.

Assignees

• Baxalta Inc
• Baxalta GmbH

Inventors

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

Classifications

• C12N5/0018 Primary

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

• C12N7/00

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

• C12N2500/10

  Metals; Metal chelators · CPC title

• C12N2500/24

  Iron; Fe chelators; Transferrin · CPC title

• C12N2710/24151

  Methods of production or purification of viral material · CPC title