Abstract
A novel method for recombinant adeno-associated virus (rAAV) purification on large scale is described. The method involves three steps, including chloroform treatment, PEG/NaCl precipitation and chloroform extraction. The whole procedure can be performed in four hours. Using this purification method, we can reproducibly obtain, from 4 × 109 of proviral cells cultured in roller bottles, purified rAAV-GFP stocks with titers of around 5 × 1013 particles/mL and purity greater than 95%. The infectious titers of the vector stocks were up to 2 × 1012 TU/mL, thus particle-to-infectivity rate was about 25. Under an electronic microscope, most rAAV particles appeared full and a few were in intermediate form. Empty particles were rarely seen. The purified rAAV-GFP stocks have been successfully used inin vitro andin vivo transfection experiments. Therefore, this new method offers a simple, rapid and cost-effective way for large-scale rAAV purification.
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Wu, X., Dong, X., Wu, Z. et al. A novel method for purification of recombinant adenoassociated virus vectors on a large scale. Chin.Sci.Bull. 46, 485–488 (2001). https://doi.org/10.1007/BF03187263
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DOI: https://doi.org/10.1007/BF03187263