Lysis, extraction and purification of adeno-associated virus and adenovirus from host cells
OwnerDao-Yao Heinventor
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Abstract
The present disclosure provides compositions and methods for extracting viral particles of a recombinant adeno-associated virus (AAV) or an adenovirus (AdV) from a sample comprising cells enclosing the viral particles. The method can include lysing the cells with the addition of an alkaline lysis solution to the sample such that the sample comprises compound(s) buffering pH value about 9.0 to about 11.5 and about 0.1% to about 1% of a detergent precipitating host cell proteins by adding a salting solution to the sample such that the sample comprises about 0.5 M to about 2.0 M of a salt and has a pH of about 4.0 to about 7.0.
Claims10
A method of extracting viral particles of a recombinant adeno associated virus (AAV) or an adenovirus (AdV) from a sample comprising cells enclosing the viral particles, comprising:
- lysing the cells by the addition of an alkaline lysis solution to the sample such that the sample comprises about 0.1% to about 1% of a detergent comprising a salt of deoxycholic acid and has a pH of about 9.5 to about 11 under suitable conditions to lyse the cells;
- then
- precipitating proteins released from the lysed cells by adding a salting solution to the sample such that the sample comprises about 0.5 M to about 2.0 M of a salt selected from the group consisting of sodium acetate, potassium acetate and combinations thereof and has a pH of about 4.0 to about 7.0;
- then
- removing the precipitated proteins to obtain a supernatant comprising the viral particles;
- and then
- precipitating the viral particles by adding a precipitation solution to the supernatant such that the supernatant comprises about 5% to about 12% of a polyethylene glycol (PEG),
- wherein the method does not include use of ultracentrifugation or freeze-thaw cycles.
Dependencies are inferred from the claim text.