Skip to Content
MilliporeSigma
  • Rapid and simplified purification of recombinant adeno-associated virus.

Rapid and simplified purification of recombinant adeno-associated virus.

Journal of virological methods (2012-05-09)
Ping Guo, Yousef El-Gohary, Krishna Prasadan, Chiyo Shiota, Xiangwei Xiao, John Wiersch, Jose Paredes, Sidhartha Tulachan, George K Gittes
ABSTRACT

Preclinical gene therapy studies both in vitro and in vivo require high purity preparations of adeno-associated virus (AAV). Current methods for purification of AAV entail the use of centrifugation over either a CsCl or iodixanol gradient, or the use of chromatography. These methods can be cumbersome and expensive, necessitating ultrahigh speed gradient centrifugation or, for chromatography the use of other expensive equipment. In addition, these methods are time consuming, and the viral yield is not high. Currently no commercial purification kits are available for other than AAV serotype 2. A simplified method was used for the purification of AAV, with a viral yield that is able to be used effectively in adult and embryo mice. The method does not require ultrahigh speed gradient centrifugation nor chromatography. Instead, polyethylene glycol (PEG)/aqueous two-phase partitioning is used to remove soluble proteins from the PEG8000 precipitated virus-protein mixture. The procedure obtained rapidly up to 95% recovery of high quality purified AAV. The entire purification process, including HEK293 cell transfection, can be completed readily within one week, with purity seemingly higher than that obtained after one round of CsCl gradient purification.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium carbonate decahydrate, 99.999% trace metals basis
Sigma-Aldrich
Sodium carbonate, BioUltra, anhydrous, ≥99.5% (calc. on dry substance, T)
Sigma-Aldrich
Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Sodium carbonate, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium carbonate monohydrate, puriss., meets analytical specification of Ph. Eur., BP, NF, FCC, E500, 99.5-100.5% (ex dried subst.)
Supelco
Sodium carbonate, reference material for titrimetry, certified by BAM, >99.5%
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Sodium carbonate decahydrate, puriss. p.a., ≥99.0% (T)
Sigma-Aldrich
Sodium carbonate, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Sodium carbonate, puriss., meets analytical specification of Ph. Eur., BP, NF, FCC, E500, anhydrous, 99.5-100.5% (calc. to the dried substance)
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent (primary standard), 99.95-100.05% dry basis
Sigma-Aldrich
Sodium carbonate, anhydrous, powder or granules, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium carbonate monohydrate, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Sodium carbonate, ACS reagent, anhydrous, ≥99.5%, powder or granules
Sigma-Aldrich
Sodium carbonate, ACS reagent (primary standard), anhydrous, 99.95-100.05% dry basis
Sigma-Aldrich
Sodium carbonate monohydrate, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium carbonate, powder, ≥99.5%, ACS reagent
Sigma-Aldrich
Sodium carbonate-12C, 99.9 atom % 12C
Supelco
Sodium carbonate concentrate, 0.1 M Na2CO3 in water, eluent concentrate for IC
Supelco
Sodium carbonate concentrate, Na2CO3 72 mM in water, IC eluent concentrate (20x) for Metrosep A Supp 7