A Simple, Two-Step, Small-Scale Purification of Recombinant Adeno-Associated Viruses.

A Simple, Two-Step, Small-Scale Purification of Recombinant Adeno-Associated Viruses. J Virol Methods. 2020 May 01;:113863 Authors: Chen SH, Papaneri A, Walker M, Scappini E, Keys RD, Martin NP Abstract Recombinant adeno-associated viruses (rAAVs) are robust and versatile tools for in vivo gene delivery. Natural and designer capsid variations in rAAVs allow for targeted gene delivery to specific cell types. Low immunogenicity and lack of pathogenesis also add to the popularity of this virus as an innocuous gene delivery vector for gene therapy. rAAVs are routinely used to express recombinases, sensors, detectors, CRISPR-Cas9 components, or to simply overexpress a gene of interest for functional studies. High production demand has given rise to multiple platforms for the production and purification of rAAVs. However, most platforms rely heavily on large amounts of starting material and multiple purification steps to produce highly purified viral particles. Often, researchers require several small-scale purified rAAVs. Here, we describe a simple and efficient technique for purification of recombinant rAAVs from small amounts of starting material in a two-step purification method. In this method, rAAVs are released into the packaging cell medium using high salt concentration, pelleted by ultracentrifugation to remove soluble impurities. Then, the resuspended pellet is purified using a protein spin-concentrator. In this protocol, we modi...
Source: Journal of Virological Methods - Category: Virology Authors: Tags: J Virol Methods Source Type: research