Method for harvesting expression products

Patent metadata

Field	Value
Publication number	US-10087423-B2
Application number	US-201113811093-A
Country	US
Kind code	B2
Filing date	Jul 15, 2011
Priority date	Jul 20, 2010
Publication date	Oct 2, 2018
Grant date	Oct 2, 2018

Abstract

The present invention provides methods and systems for recovering an essentially cell-associated expression product from a host cell comprising (a) culturing said host cell under conditions that allow expression of said expression product; (b) collecting said host cell in/on a filter unit; (c) disrupting said host cell in/on the filter unit; and (d) separating said expression product from said disrupted host cell. The present invention also provides an expression product obtainable by the methods or systems, wherein the expression product is a poxvirus.

First claim

The invention claimed is:

1. A method for recovering a poxvirus from a host cell comprising
a. culturing the host cell under conditions that allow expression of the poxvirus;
b. collecting the host cell in or on a filter unit;
c. disrupting the host cell retained in or on the filter unit; and
d. separating the poxvirus from the disrupted host cell.

2. The method of claim 1 , wherein the poxvirus is selected from the group consisting of fowlpox virus, vaccinia virus, and modified vaccinia virus Ankara (MVA).

3. The method of claim 2 , wherein the filter unit comprises a pore size of less than 10 μm.

4. The method of claim 1 , wherein the poxvirus is a recombinant poxvirus.

5. The method of claim 1 , wherein the host cell is a vertebrate or an insect cell.

6. The method of claim 5 , wherein the vertebrate cell is an avian cell.

7. The method of claim 5 , wherein the host cell is cultured in suspension culture.

8. The method of claim 5 , wherein the host cell is cultured in a disposable bioreactor.

9. The method of claim 8 , wherein the disposable bioreactor is a wave bioreactor.

10. The method of claim 1 , wherein the host cell is disrupted in or on the filter unit by lysis.

11. The method of claim 10 , wherein the lysis is hypotonic lysis.

12. The method of claim 1 , wherein the expression product is eluted for recovery.

13. The method of claim 1 , wherein the filter unit for collecting the host cell is a depth filter.

14. The method of claim 1 , further comprising a step of purifying the recovered expression product.

15. The method of claim 14 , wherein the purifying step includes a chromatography step selected from the group consisting of hydrophobic interaction chromatography, pseudo-affinity chromatography, anion exchange chromatography and size exclusion chromatography.

16. The method of claim 15 , further comprising a step of formulating the recovered expression product with a pharmaceutically acceptable carrier.

Assignees

• Wehnes Engelbert
• Werner Uwe
• Leschke Christian
• Bavarian Nordic As

Inventors

• Wehnes Engelbert
• Werner Uwe
• Leschke Christian

Classifications

• A61P31/20

  for DNA viruses · CPC title

• C12N2710/24151

  Methods of production or purification of viral material · CPC title

• C12N7/02 Primary

  Recovery or purification · 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

• C12M47/02 Primary

  Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02) · CPC title