Patent ID: 11878056
Assignee: THE CHILDREN'S HOSPITAL OF PHILADELPHIA
Field: Pharmaceuticals (Chemistry)
Classification: CPC A  C | IPC A  C

Claim 0:
1. A method for producing a highly purified adeno-associated virus (AAV) vector formulation said method comprising the steps of:
(a) harvesting cells comprising recombinant AAV vector particles;
(b) concentrating said cells harvested in step (a) via tangential flow filtration to produce a concentrated harvest;
(c) lysing said concentrated harvest produced in step (b) to produce a lysate;
(d) clarifying 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 diafiltration;
(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.