Small-Scale Recombinant Adeno-Associated Virus Purification

Part of the Methods in Molecular Biology book series (MIMB, volume 1382)

Abstract

Recombinant adeno-associated virus (rAAV) vectors have become increasingly popular in research and clinical trials due to their efficient gene transfer and long-term expression in tissues including brain. In addition, rAAV has demonstrated an impressive safety profile in gene therapy trials. The emergence of rAAV serotypes with different cell tropisms and distribution properties has allowed scientists to tailor serotypes to specific experimental needs. AAV does not have a cytopathic effect; therefore, purification methods require extraction of the viral vector from the cell. This involves gradient ultracentrifugation of the cellular extract sometimes followed by chromatography. This chapter describes a small-scale production method for rAAV purification from ten to twenty 15 cm plates of human embryonic kidney-derived 293B cells (HEK 293) cells that can yield approximately 300 μl of a 5 × 1012 to 1 × 1013 genome copies/ml viral preparation final concentration.

Key words

Adeno-associated virus Iodixanol gradient purification Tissue culture 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Neurology, Medical Sciences CenterUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Molecular Pharmacology and Physiology, Byrd Alzheimer Institute, College of MedicineUniversity of South FloridaTampaUSA

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