Chromatography-based purification of adeno-associated virus.
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.
Smith
Richard H
RH
Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD, USA.
Yang
Linda
L
Kotin
Robert M
RM
D015153:Blotting, Western; D002850:Chromatography, Gel / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000302:isolation & purification*; D006801:Humans