Patent ID: 9408904
Filing Date: 2016-08-09
CPC Classification: A61K,A61P,C07K,C12N,C12Y

Claim Text:
1. A method for producing a highly purified adeno-associated (AAV) vector formulation, said method comprising the steps of: (a) harvesting cells and cell culture supernatant comprising recombinant AAV vector particles; (b) concentrating said cells and said cell culture supernatant harvested in step (a) via tangential flow filtration to produce a concentrated harvest; (c) lysing said concentrated harvest produced in step (b) by microfluidization to produce a lysate; (d) filtering said lysate produced in step (c) to produce a clarified lysate; (e) subjecting said clarified lysate produced in step (d) to ion exchange column chromatography to produce a column eluate comprised of purified AAV vector particles, and optionally concentrating said column eluate by tangential flow filtration to produce a concentrated column eluate; (f) mixing said column eluate or said concentrated column eluate produced in step (e) with cesium chloride to produce a mixture, and subjecting said mixture to gradient ultracentrifugation to substantially separate said bona fide AAV vector particles from empty capsid AAV vector particles and other AAV vector related impurities; (g) collecting said bona fide AAV vector particles separated in step (f) and subjecting said collected bona fide AAV vector particles to a buffer exchange by tangential flow filtration; (h) formulating said bona fide AAV vector particles resulting from step (g) with surfactant to produce an AAV vector formulation; and (i) filtering said AAV vector formulation produced in step (h) to produce a highly purified AAV vector formulation in which at least 90% of the AAV vector particles in said highly purified AVV vector formulation are bona fide AAV particles.