A novel method for purification of recombinant adenoassociated virus vectors on a large scale

A novel method for recombinant adeno-associated virus (rAAV) purification o n large scale is described. The method involves three steps, including chlo roform treatment, PEG/NaCl precipitation and chloroform extraction, The who le procedure can be performed in four hours. Using this purification method , we can reproducibly obtain, from 4 X 10(9) of proviral cells cultured in roller bottles, purified rAAV-GFP stocks with titers of around 5 X 10(13) p articles/mL and purity greater than 95%. The infectious titers of the vecto r stocks were up to 2 X 10(12) TU/mL, thus particle-to-infectivity rate was about 25. Under an electronic microscope, most rAAV particles appeared ful l and a few were in intermediate form. Empty particles were rarely seen. Th e purified rAAV-GFP stocks have been successfully used in in vitro and in v ivo transfection experiments, Therefore, this new method offers a simple, r apid and cost-effective way for large-scale rAAV purification.