Summary
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.
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Acknowledgements
This research was supported by the Intramural Research Program of the National Heart, Lung, and Blood Institute, National Institutes of Health. We thank Sylvain Cecchini for critical review of the draft manuscript.
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Smith, R.H., Yang, L., Kotin, R.M. (2008). Chromatography-Based Purification of Adeno-Associated Virus. In: Le Doux, J.M. (eds) Gene Therapy Protocols. Methods in Molecular Biology™, vol 434. Humana Press. https://doi.org/10.1007/978-1-60327-248-3_4
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DOI: https://doi.org/10.1007/978-1-60327-248-3_4
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