US 11,840,710 B2
Methods for purification of recombinant AAV vectors
Paulene McLean Quigley Sheldon, Seattle, WA (US); Peter S. Gagnon, San Clemente, CA (US); Gina Nichols, Everett, WA (US); and Barbara A. Thorne, Sammamish, WA (US)
Assigned to Genzyme Corporation, Cambridge, MA (US)
Filed by Genzyme Corporation, Cambridge, MA (US)
Filed on Sep. 28, 2020, as Appl. No. 17/035,480.
Application 17/035,480 is a continuation of application No. 16/878,375, filed on May 19, 2020, abandoned.
Application 16/878,375 is a continuation of application No. 16/006,210, filed on Jun. 12, 2018, granted, now 10,696,952, issued on Jun. 30, 2020.
Application 16/006,210 is a continuation of application No. 13/328,306, filed on Dec. 16, 2011, granted, now 10,017,746, issued on Jul. 10, 2018.
Application 13/328,306 is a continuation of application No. PCT/US2010/038897, filed on Jun. 16, 2010.
Claims priority of provisional application 61/187,601, filed on Jun. 16, 2009.
Prior Publication US 2021/0139860 A1, May 13, 2021
Int. Cl. C12N 7/00 (2006.01); C07K 14/005 (2006.01); C12N 15/86 (2006.01)
CPC C12N 7/00 (2013.01) [C07K 14/005 (2013.01); C12N 15/86 (2013.01); C12N 2750/14151 (2013.01)] 24 Claims
 
1. A method for isolating a population of recombinant adeno-associated virus (rAAV) particles from in-process impurities in a feedstream, comprising the steps of:
(a) subjecting the feedstream containing the rAAV particles to flow-through anionic exchange filtration and/or subjecting the feedstream containing the rAAV particles to tangential flow filtration (TFF);
(b) contacting the feedstream containing the rAAV particles with an apatite chromatography medium in the presence of polyethylene glycol (PEG), wherein the rAAV particles bind to the apatite chromatography medium; and
(c) eluting the rAAV particles bound to the apatite chromatography medium with an elution buffer containing less than 3% (w/v) PEG; and
(d) heat inactivating a helper virus in the feedstream eluted from the apatite chromatography medium, wherein the heat inactivation step comprises heating the feedstream eluted from the apatite chromatography medium to 52° C.