One-step AAV particle isolation: AAVanced

The AAVanced™ Concentration Reagent from System Biosciences offers a simple, one-step protocol to concentrate AAV particles from media to streamline gene therapy development. Specifically optimised for the precipitation of most AAV serotypes from culture medium and based on an established nanoparticle technology, the reagent significantly reduces the complexity and time required for rAAV virus production. rAAV particles produced using this reagent have been tested successfully for multiple serotypes of AAV, with no observed cytotoxic effects, making it ideal for challenging in vitro and in vivo applications.

Benefits of using AAVanced Concentration Reagent
• Easy to use, single reagent
• Isolate rAAV directly from media
• No cell lysates or centrifugation
• Compatible with all serotypes

How it works

rAAV particles isolated using AAVanced Concentration Reagent are present at high copy numbers in 293T cells

A small volume of the rAAV virus isolated using AAVanced Concentration Reagent was used to infect 3×105 293T cells in a 24-well culture dish. The cells were imaged after 72-hours and rAAV copy numbers quantitated by qPCR. The copy numbers measured for the PGK-GFP rAAV virus were quite high at 2×1013 copies/mL.

AAVanced Concentration Reagent works with multiple serotypes in vitro

The same rAAV shuttle vector with a PGK promoter driving GFP expression was packaged in parallel into three different AAV serotypes: AAV-2, AAV-DJ and AAV-5. After 48-hours, media was collected and rAAV particles isolated using the AAVanced Concentration Reagent. Each rAAV particle pellet was resuspended in 100 mL of sterile PBS, and then 1 mL of the rAAV suspensions were added to three different cell lines (HEK293T, HT1080 and CHO) to validate transduction efficiencies. The infected cells were imaged for GFP expression 6-days after addition of the isolated rAAV particles. All serotypes of rAAV were able to infect the three different cell types tested, demonstrating the broad compatibility of AAVanced Concentration Reagent.

Material available for download
AAVanced Concentration Reagent product sheet