Removal of contaminating viruses from AAV preparations

Patent metadata

Field	Value
Publication number	US-9840694-B2
Application number	US-201214343884-A
Country	US
Kind code	B2
Filing date	Sep 7, 2012
Priority date	Sep 8, 2011
Publication date	Dec 12, 2017
Grant date	Dec 12, 2017

Abstract

The present invention relates to a separation of viruses of an essentially spherical shape from viruses with a rod-like shape that are comprised in a sample, wherein the sample comprising the viruses is subjected to filtration.

First claim

The invention claimed is:

1. A method for removing baculoviral virions from a sample comprising parvoviral virions and baculoviral virions, which method comprises filtering the sample through a filter that has a pore size of 30-40 nm, thereby removing the baculoviral virions from the sample.

2. The method according to claim 1 , wherein the parvoviral virions are adeno-associated virus virions.

3. The method according to claim 1 , wherein the baculoviral virions being removed are virions of Autographa californica multicapsid nucleopolyhedrovirus.

4. The method according to claim 1 , wherein the filter is a virus filter or an ultrafilter.

5. The method according to claim 1 , wherein the filter is a surface and/or a depth filter.

6. The method according to claim 1 , wherein the filter comprises a material selected from the group consisting of: regenerated cellulose, polyvinylidene fluoride, polysulfone, polytetrafluoroethylene, polypropylene, modified polyethersulfone, unmodified polyethersulfone, cellulose acetate, cellulose nitrate, and polyamide.

7. The method according to claim 1 , wherein the filter has a pore size of 32-38 nm.

8. The method according to claim 6 , wherein the regenerated cellulose filter is a cuprammonium-regenerated cellulose filter.

9. The method according to claim 1 , wherein the sample is filtered through two or more filters.

10. The method according to claim 1 , wherein prior to the filtering step, the sample is prefiltered through a filter with a pore size of 70-200 nm.

11. The method according to claim 1 , wherein the pH of the sample is 6 to 10.

12. The method according to claim 1 , wherein 1 ml to 200 ml of the sample is filtered per cm 2 of filter surface area.

13. The method according to claim 1 , wherein the filtering results in:
(a) recovery of at least 85% of the parvoviral virions; and
(b) at least a 10 5 -fold reduction in baculoviral virion titer.

14. The method according to claim 8 wherein the cuprammonium-regenerated cellulose filter is a hollow fiber filter.

15. The method according to claim 6 wherein the filter is a hollow fiber membrane comprising cuprammonium regenerated cellulose, or a membrane comprising hydrophilic modified polyvinylidenefluoride.

16. The method according to claim 11 wherein the sample pH is 7 to 9.

17. The method according to claim 11 wherein the sample pH is 7.5 to 8.5.

18. The method according to claim 11 wherein the sample pH is about 8.0.

19. The method according to claim 12 wherein 80 to 120 ml of the sample are filtered per cm 2 of the filter surface area.

Assignees

• Hermens Wilhelmus Theodorus Johannes Maria Christiaan
• Smith James Patrick
• Uniqure Ip Bv

Inventors

• Hermens Wilhelmus Theodorus Johannes Maria Christiaan
• Smith James Patrick

Classifications

• C12N7/02

  Recovery or purification · CPC title

• C12N7/00 Primary

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

• B01D2311/2649

  Filtration · CPC title

• B01D61/145

  Ultrafiltration · CPC title

• B01D61/16

  Feed pretreatment · CPC title