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Briefings in Functional Genomics and Proteomics Advance Access originally published online on July 16, 2008
Briefings in Functional Genomics and Proteomics 2008 7(4):303-311; doi:10.1093/bfgp/eln034
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Published by Oxford University Press 2008.

Strategies for manufacturing recombinant adeno-associated virus vectors for gene therapy applications exploiting baculovirus technology

Alejandro Negrete and Robert M. Kotin

Corresponding author. Robert M. Kotin, Bld. 10, Rm. 7D05, 10 Center Dr. Bethesda, MD 20892, USA. Tel: (001) 301-496-1594; E-mail: kotinr{at}nhlbi.nih.gov

The development of recombinant adeno-associated virus (rAAV) gene therapy applications is hampered by the inability to produce rAAV in sufficient quantities to support pre-clinical and clinical trials. Contrasting with adherent cell cultures, suspension cultures provide a straightforward means for expansion, however, transiently expressing the necessary, but cytotoxic virus proteins remains the challenge for rAAV production. Both the expansion and expression issues are resolved by using the baculovirus expression vector (BEV) and insect cell culture system. This review addresses strategies for the production of rAAV exploiting baculovirus technology at different scales using different configurations of bioreactors as well as processing and product characterization issues. The yields obtained with these optimized processes exceed ~1 x 1014 vector particles per liter of cell culture suitable for pre-clinical and clinical trials and possible commercialization.

Keywords: adeno-associated vectors, gene therapy, large-scale production, baculovirus, insect cell


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