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AAVanced™ Concentration Reagent

One-step rAAV particle isolation

Easiest method for rAAV isolation

  • Easy to use, single reagent
  • Isolate rAAV directly from media
  • No cell lysates or centrifugation
  • Saves time and cost-effective
  • Compatible with all serotypes

AAVanced™ Concentration Reagent

One-step rAAV particle isolation from packaging cell media
Traditional rAAV production methods requires multiple steps, including cell lysis and CsCl2 ultra-high-speed density gradient centrifugation, chromatography, or binding to affinity matrix columns. These are difficult to setup, time-consuming, and require specialized equipment for isolation of high-purity rAAV for in vivo experiments. As many studies have shown that different rAAV serotypes are efficiently secreted into the culture medium of transfected 293T cells during rAAV packaging, SBI has developed a novel reagent to facilitate fast, high-quality rAAV isolation.

   

Easiest Protocol to Isolate rAAV Particles

SBI has developed an innovative solution called AAVanced Concentration Reagent - a simple, one step rAAV concentration reagent for the isolation of rAAV particles from media. AAVanced Concentration Reagent is based on a proven nanoparticle technology and is specifically optimized for the precipitation of most AAV serotypes from culture medium. The rAAV virus produced using AAVanced Concentration Reagent has been tested successfully for in vitro and in vivo applications for multiple serotypes of AAV with no observed cytotoxic effects, which provides direct evidence for the utility of this reagent for demanding applications using rAAV. SBI's AAVanced Concentration Reagent significantly reduces the complexity and time required for rAAV virus production, allowing researchers to focus on their research experiments rather than virus production.

AAVanced Concentration Reagent Produces High Titers of Robust Virions

High titer rAAV generated with the AAVanced concentration reagent
SBI supplied rAAV paricles harboring a PGK-GFP shuttle vector and virus concentrated from HEK293 packaging cell media to determine virus titers. A small volume of the rAAV virus (0.1 ul) was used to infect 3x10^5 293T test 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 2x10^13 Genome Copies/ml. The cell infection image data are shown below.

AAVanced virus produces high titer virus

How does AAVance virus preps compare to tranditonal cell lysate preps?
To test this comparison, recombinant rAAV was packaged with a PGK-GFP expression shuttle vector. The rAAV particles were isolated either by the traditional cell lysis freeze-thaw with centifugation method or with the AAVanced Concentration Reagent from the packaging cell media. Equal amounts of rAAV were then added to HEK293 cells to test for infectivity based on GFP expression. Representative images of GFP expressing cells 10 days after infection.

AAVanced virus produces infectious virus