Chromatography-based purification of adeno-associated virus

Methods Mol Biol. 2008:434:37-54. doi: 10.1007/978-1-60327-248-3_4.

Abstract

Virus-mediated gene transfer shows great potential as a therapeutic strategy for the management of various inherited and acquired human diseases. Among the current viral vectors, adeno-associated virus (AAV) has become the vector of choice for numerous gene therapy applications. As AAV-based vectors approach the clinic, the need for scalable methods of production and purification is steadily increasing. In this chapter, we present a column chromatography-based protocol for the purification of recombinant AAV type 1 (AAV-1) to near homogeneity. The protocol, which can be completed within one working day, employs three major purification steps: (1) polyethylene glycol-mediated vector precipitation, (2) anion-exchange chromatography, and (3) gel filtration chromatography. This method provides a basic strategy, or "platform," that can be adapted to the purification of other recombinant AAV vector serotypes.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Blotting, Western
  • Chromatography, Gel / methods*
  • Chromatography, Ion Exchange / methods*
  • Dependovirus / genetics*
  • Genetic Vectors / isolation & purification*
  • Humans