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US 2004/0005693 A1Published application
Viral purification methods
AssigneeOncolytics Biotech Inc.
InventorMatthew C. Coffey
Life of this patent
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Abstract
The present invention is directed to an improved method of purifying virus, particularly reovirus. Infectious virus can be extracted from a cell culture with a detergent to produce high titers of virus, and the virus can then be purified by simple steps such as filtration and column chromatography. Viruses and compositions comprising the viruses prepared according to the present invention are also provided.
Claims23
A method of producing virus from a culture of cells, comprising the steps of:
- (a) providing a culture of cells which has been infected by the virus;
- (b) extracting the virus from the cells by adding a detergent to the culture and incubating for a period of time to result in a cell lysate;
- (c) removing cell debris;
- and
- (d) collecting the virus.
2 more elements
A method of producing infectious reovirus, comprising:
- (a) providing a culture of HEK 293 cells which has been infected by reovirus;
- (b) extracting the virus from the cells by adding Triton X-100 to the culture and incubating at about 25° C. to about 37° C.;
- (c) treating the mixture from step (b) with benzonase;
- (d) removing cell debris by filtration;
- (e) concentrating the filtrate by ultrafiltration or diafiltration;
- (f) purifying the reovirus by a combination of ion exchange and size exclusion chromatography;
- and
- (g) collecting the reovirus.
5 more elements
Dependencies are inferred from the claim text.