Scalable purification method for aav9
AssigneeThe Trustees of the University of Pennsylvania
InventorsMartin Lock, Mauricio Alvira
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Abstract
A two-step chromatography purification scheme is described which selectively captures and isolates the genome-containing rAAV vector particles from the clarified, concentrated supernatant of a rAAV production cell culture. The process utilizes an affinity capture method performed at a high salt concentration followed by an anion exchange resin method performed at high pH to provide rAAV vector particles which are substantially free of rAAV intermediates.
Claims1
A method for separating AAV9 viral particles having packaged genomic sequences from genome-deficient AAV9 capsid intermediates, said method comprising:
- subjecting a mixture comprising recombinant AAV9 viral particles and AAV9 capsid intermediates to fast performance liquid chromatography, wherein the AAV9 viral particles and AAV9 intermediates are bound to an anion exchange resin equilibrated at a pH of about 10.2 and subjected to a salt gradient while monitoring eluate for ultraviolet absorbance at about 260 and about 280, wherein the AAV9 full capsids are collected from a fraction which is eluted when the ratio of A260/A280 reaches an inflection point.
Dependencies are inferred from the claim text.