Medium for the protein-free and serum-free cultivation of cells

Patent metadata

Field	Value
Publication number	US-9982286-B2
Application number	US-201615235453-A
Country	US
Kind code	B2
Filing date	Aug 12, 2016
Priority date	Sep 28, 1999
Publication date	May 29, 2018
Grant date	May 29, 2018

Abstract

A medium is described for the protein-free and serum-free cultivation of cells, especially mammalian cells, whereby the medium contains a proportion of soy hydrolysate.

First claim

What is claimed is:

1. A method for increasing productivity of a recombinant protein in a cell culture, the method comprising:
culturing mammalian cells that express the recombinant protein in an animal protein-free and serum-free cell culture medium; and
adding more than 10 wt. % ultrafiltered soy hydrolysate based on the total dry weight of the medium to the animal protein-free and serum-free cell culture medium,

wherein at least 40% of the soy hydrolysate has a molecular weight of ≤500 daltons.

2. The method of claim 1 , wherein at least 50% of the soy hydrolysate has a molecular weight of ≤500 daltons.

3. The method of claim 1 , wherein at least 55% of the soy hydrolysate has a molecular weight of ≤500 daltons.

4. The method of claim 1 , wherein the soy hydrolysate has an endotoxin content of <500 U/g.

5. The method of claim 1 , wherein the mammalian cells are CHO cells or BHK cells.

6. The method of claim 1 , wherein the recombinant protein is selected from the group consisting of: Factor II, Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Protein S, Protein C, activated forms of these factors, and vWF.

7. The method of claim 6 , wherein the recombinant protein is Factor VIII.

8. The method of claim 1 , wherein the medium comprises an amino acid.

9. The method of claim 8 , wherein the amino acid is selected from the group consisting of L-asparagine, L-cysteine, L-cystine, L-proline, L-tryptophan, L-glutamine and mixtures thereof.

10. The method of claim 1 , wherein the medium further comprises: 1 to 100 g/L synthetic minimal medium; 0.05-1 g/L glutamine; 0.0005-0.05 g/L ascorbic acid; 0.1-10 g/L NaHCO 3 ; 0.0005-0.05 g/L ethanolamine; and 1-15 μg/L sodium selenite.

11. The method of claim 1 , wherein the cells are cultured in a bioreactor.

Assignees

• Baxalta Inc
• Baxalta GmbH

Inventors

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

Classifications

• C12N2500/76

  from plants · CPC title

• C12N5/0037

  Serum-free medium, which may still contain naturally-sourced components · CPC title

• C12N5/005

  Protein-free medium · CPC title

• C12N2500/46

  Amines, e.g. putrescine · CPC title

• C12N2500/05

  Inorganic components · CPC title