Process for protein production

Patent metadata

Field	Value
Publication number	US-10138290-B2
Application number	US-201113823847-A
Country	US
Kind code	B2
Filing date	Oct 5, 2011
Priority date	Oct 5, 2010
Publication date	Nov 27, 2018
Grant date	Nov 27, 2018

Abstract

The present invention relates to a process for the production of a haemostasis protein by continuous perfusion culturing of a cell culture in suspension, said cell culture expressing said haemostasis protein into said culture suspension, wherein the cell culture flows across a filter module, which filter module leads to a harvest port, the filter module having a mesh size of from 0.1 to 2.9 μm allowing passage across of the haemostasis protein and wherein the flow across the filter module is an alternating tangential flow. The invention also relates to a protein produced by the process of the invention.

First claim

The invention claimed is:

1. A large-scale, continuous perfusion process for producing a haemostasis protein, comprising
providing a cell culture in suspension in a bioreactor, wherein the bioreactor is in fluid communication with a filter module, wherein the bioreactor volume is at least 500 L, and wherein the filter module mesh size is from about 0.1 μm to about 2.9 μm;
providing a dissolved oxygen concentration in the suspension of around 20% to 100%; and
flowing the cell culture suspension across the filter module in an alternating tangential flow, in order to separate haemostasis protein produced by the cell culture from the cells; and
culturing the cells to a target cell density below 80×10 6 cell s/ml.

2. The process of claim 1 , wherein the haemostasis protein is Factor IX.

3. The process of claim 1 , wherein the haemostasis protein is Factor VIII.

4. The process of claim 1 , wherein the haemostasis protein is Factor VII.

5. The process of claim 1 , wherein the perfusion rate is from 0.7 to 10 times the bioreactor volume per day.

6. The process of claim 1 , wherein unfiltered cell culture suspension is removed at a rate of 0 to 50% of the bioreactor volume per day.

7. The process of claim 6 , wherein unfiltered cell culture suspension is removed at a rate of 1 to 20% of the bioreactor volume per day.

8. The process according to claim 1 , further comprising removing unfiltered cell culture suspension continuously.

9. The process according to claim 1 , further comprising removing unfiltered cell culture suspension discontinuously.

10. The process according to claim 1 , further comprising culturing the cells to a target cell density below 50×10 6 cells/ml.

11. The process according to claim 5 , wherein the perfusion rate is from 0.9 to 4 times the bioreactor volume per day.

12. The process according to claim 1 , wherein the cells in the cell suspension are Chinese Hamster Ovary (CHO) cells.

13. The process according to claim 12 , wherein the cells in the cell suspension are CHO-KI cells.

14. The process according to claim 1 , wherein the dissolved oxygen concentration is around 30 to 70%.

15. The process according to claim 14 , wherein the dissolved oxygen concentration is around 45 to 55%.

16. The process according to claim 15 , wherein the dissolved oxygen concentration is around 50%.

Assignees

• Robin Jarno
• Novo Nordisk Healthcare Ag

Inventors

• Robin Jarno

Classifications

• C12N9/644

  Coagulation factor IXa (3.4.21.22) · CPC title

• C12Y304/21021

  Coagulation factor VIIa (3.4.21.21) · CPC title

• C07K14/755 Primary

  Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)} · CPC title

• C12N9/6437

  Coagulation factor VIIa (3.4.21.21) · CPC title

• C12Y304/21022

  Coagulation factor IXa (3.4.21.22) · CPC title