US 10,889,797 B2
Three phase partitioning (TPP) method for virus purification
Haifeng Chen, Piedmont, CA (US)
Assigned to Virovek Incorporation, Hayward, CA (US)
Filed by Virovek, Inc., Hayward, CA (US)
Filed on Nov. 28, 2018, as Appl. No. 16/203,055.
Claims priority of provisional application 62/591,383, filed on Nov. 28, 2017.
Prior Publication US 2019/0161725 A1, May 30, 2019
Int. Cl. C12N 1/02 (2006.01); C12N 1/04 (2006.01); C12N 5/07 (2010.01); C12N 1/06 (2006.01); C12N 7/00 (2006.01)
CPC C12N 1/02 (2013.01) [C12N 1/04 (2013.01); C12N 1/06 (2013.01); C12N 5/0601 (2013.01); C12N 7/00 (2013.01); C12N 2750/14151 (2013.01)] 24 Claims
1. A method of purifying an adeno-associated virus from a baculovirus-infected cell lysate, the method comprising:
a) forming a first mixture comprising i) an aqueous phase comprising water and a cell lysate comprising an adeno-associated virus, ii) t-butanol, and iii) ammonium sulfate at a first concentration;
b) separating the first mixture to form a first organic phase, a first interphase, and a first aqueous phase; and
c) collecting the first aqueous phase, wherein the ammonium sulfate at a first concentration is ammonium sulfate at a concentration at which t-butanol is immiscible with water.