AAV production by transient transfection: strategies & challenges

AAV is the most common modality for gene therapy, with five approved drugs and over 200 in development. The most widely used method to manufacture AAV is by transient transfection of HEK293 cells. This review will describe strategies, challenges, and recent advances in the upstream cell culture and transfection operations of producing AAV. These strategies are categorized into four areas that approximate the primary raw materials of the process: transfection reagent, plasmids, medium, and cell line. Although we examine each category independently, we emphasize the importance of parallel development and combinatorial evaluation. 

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