Use of low temperature and/or low pH in cell culture

Patent metadata

Field	Value
Publication number	US-9988662-B2
Application number	US-10726408-A
Country	US
Kind code	B2
Filing date	Apr 22, 2008
Priority date	Apr 23, 2007
Publication date	Jun 5, 2018
Grant date	Jun 5, 2018

Abstract

The present disclosure provides a method of reducing protein misfolding and aggregation in the cell culture by growing the cell culture at a reduced temperature and/or reduced pH. As a result, the quality of the protein produced in the cell culture is greatly improved. Thus, the present disclosure facilitates improvements in the efficacy of therapeutic proteins produced in cell culture.

First claim

What is claimed is:

1. A method of improving protein production in a cell culture comprising:
(a) growing cells in the cell culture at a reduced temperature, wherein the reduced temperature is in a range of 27.0° C. to less than 30.0° C.; and
(b) growing cells in the cell culture at a reduced pH, wherein the reduced pH is in a range of 6.80 to less than 7.00;
wherein the cell culture is a mammalian cell culture;
wherein the method reduces production of misfolded proteins and/or aggregated proteins; and
wherein the protein produced is TNFR-Fc or sIL-13R.

2. The method of claim 1 , wherein the cell culture is a large-scale cell culture.

3. The method of claim 1 , wherein the cell culture is a fed-batch cell culture.

4. The method of claim 1 , wherein the cell culture is a CHO cell culture.

5. The method of claim 1 wherein the reduced temperature is 29.5° C.

6. The method of claim 1 , wherein the reduced pH is 6.95.

7. The method of claim 1 , wherein the method modifies the level of protein glycosylation of the produced protein.

8. The method of claim 7 , wherein the protein glycosylation is N-glycan sialylation.

9. The method of claim 1 , further comprising a step of isolating the protein from the cell culture.

10. The method of claim 9 , wherein the protein is further purified or processed for formulation.

11. The method of claim 10 , wherein the protein is formulated into a pharmaceutical composition.

Assignees

• Gomes Jose Manuel
• Hiller Gregory Walter
• Wyeth Llc

Inventors

• Gomes Jose Manuel
• Hiller Gregory Walter

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P37/02

  Immunomodulators · CPC title

• C12P21/02 Primary

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12N2500/33

  other than alpha-amino carboxylic acids, e.g. beta-amino acids, taurine · CPC title

• C12P21/005

  Glycopeptides, glycoproteins · CPC title