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Purification of Adeno-Associated Virus serotypes 1-9 through ion-exchange chromatography; a direct comparison to Cesium Chloride purification SM Soltys, JE Rabinowitz* Department of Medicine, Center for Translational Medicine, Thomas Jefferson University, Philadelphia PA
In this article we describe an efficient, inexpensive means of producing high titer, highly purified AAV through the use of iodixanol and ion exchange chromatography. Recombinant adeno-associated virus is rapidly emerging as the preeminent vector for gene therapy. The therapeutic use of AAV is currently being studied in a variety of diseases and its use as a vector is currently in various stages of clinical trials. The usefulness of AAV as a vector has become much more promising after the discovery of more tissue specific serotypes of the virus, which allows a vector to be designed and targeted to treat a particular disease. As the clinical use of AAV grows, the necessity of highly purified virus becomes more prevalent. Using iodixanol and ion exchange chromatography, we were able to purify AAV serotypes 1-9 with a yield comparable to purification using the commonly used Cesium Chloride method. Compared to virus purified by CsCl, the virus purified by iodixanol / chromatography using the ion exchange column Q-sepharose had a T.U. / particle ratio that was lower in most cases, meaning that less virus was necessary to transduce the same amount of cells. Purification by iodixanol / chromatography required less time and effort, produced much less waste, and generated viruses that were of higher purity.
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