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Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System - a 2 minute Preview of the Experimental Protocol
Md Nasimuzzaman, Sophia Villaveces, Johannes C. M. van der Loo, Sivani Alla
Cincinnati Children's Hospital Medical Center, Division of Experimental Hematology and Cancer Biology; University of Cincinnati College of Medicine,; The Children's Hospital of Philadelphia, Raymond G. Perelman Center for Cellular and Molecular Therapeutics;
In this protocol, AAV2 vector is produced by co-culturing Spodoptera frugiperda (Sf9) insect cells with baculovirus (BV)-AAV2-green fluorescent protein (GFP) or therapeutic gene and BV-AAV2-rep-cap infected Sf9 cells in suspension culture. AAV particles are released from the cells using detergent, clarified, purified by affinity column chromatography, and concentrated by tangential flow filtration.
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