Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column

One of the most promising gene transfer vectors in human clinical trials is AAV2. The quality of the vector preparations is a key element in obtaining reliable and reproducible data in preclinical studies. However, establishe d protocols either result in impure, low infectious virus (CsCl2 gradient c entrifugation) or demand a high level of manual and technical skills (CsCl2 gradient centrifugation, iodixanol/heparin or HPLC purification). In this study, we present an easy-to-do single-step column purification (SSCP) of A AV2 by gravity flow based on its affinity to heparin, without ultracentrifu gation. Various vector preparations generated by our method reproducibly sh owed high titers, infectivity, and purity. In vivo, our single-step column- purified AAV2 vectors mediate significantly higher transduction efficiency compared with conventional protocols. Investigators still unsatisfied with previously published techniques or new to the field of AAV production may f ind in our method an interesting alternative.