Springer Protocols: Abstract: 4. Chromatography-Based Purification of Adeno-Associated Virus

4. Chromatography-Based Purification of Adeno-Associated Virus

By: Richard H. Smith³, Linda Yang³, Robert M. Kotin³

Abstract

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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.

Affiliation(s): (3) Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, Bethesda, MD

Book Title: Gene Therapy Protocols: Design and Characterization of Gene Transfer Vectors

Series: Methods in Molecular Biology | Volume: 434 | Pub. Date: Mar-07-2008 | Page Range: 37-54 | DOI: 10.1007/978-1-60327-248-3_4

Subject: Genetics/Genomics

Key Words: AAV-1 - adeno-associated virus - PEG precipitation - chromatography - purification

References

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