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Economized large-scale production of high yield of rAAV for gene therapy applications exploiting baculovirus expression system by Alejandro Negrete; Linda C. Yang; Andres F. Mendez; Justin R. Levy; Robert M. Kotin is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract ## Background The versatility of recombinant adeno‐associated vector (rAAV) as a gene delivery system is due to the vector's ability to transduce different cell types as well as dividing and non‐dividing cells. Large‐scale production of rAAV remains one of the major challenges for continued development of pre‐clinical and clinical studies, and for its potential commercialization. The baculovirus expression vectors (BEVS) and insect cells represent a potential method to produce rAAV economically at large scale. This technology uses three different BEVs (Bac‐Rep, Bac‐GFP, and Bac‐VP) each at a multiplicity of infection (MOI) of 3. We reported previously the production of rAAV at 40 L scale using a stirred‐tank bioreactor (STB). However, production in larger volumes is limited by the stability of the BEVs and amount of BEVs needed to achieve the target MOI of 3 per BEV. Here, the production parameters were optimized and the baculovirus stability was determined. ## Methods The stability of the three types of baculovirus used to produce rAAV was determined for six expansion passages by protein expression analysis. To economize baculovirus, MOI and cell density at time of inf

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Author

Alejandro Negrete; Linda C. Yang; Andres F. Mendez; Justin R. Levy; Robert M. Kotin

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 1099-498X)

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)