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. 2024:2829:217-226.
doi: 10.1007/978-1-0716-3961-0_15.

Recombinant AAV Purification

Pranav R H Joshi  1 , Alina Venereo-Sanchez  2
Affiliations

Recombinant AAV Purification

Pranav R H Joshi et al. Methods Mol Biol. 2024.

Abstract

Purification of rAAV is a crucial unit operation of the AAV production process. It enables the capture of AAV and removal of contaminants such as host cell proteins, host cell DNA, and other cell culture-related impurities. Here we describe the purification of rAAV produced in insect cells Sf9/rBEV by immuno-affinity capture chromatography. The method is fully scale-amenable unlike other traditional purification methods based on ultracentrifugation. The method reported herein has two main steps: (1) the clarification of cell lysate by depth filtration and (2) the selective capture and single-step purification of AAV via immune-affinity chromatography. This purification method has been successfully implemented to purify the majority of wild-type AAV serotypes.

Keywords: AAV purification; Clarification; Immune-affinity chromatography; Insect cell culture; Recombinant adeno-associated virus; rBEV.

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