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PLoS One. 2012;7(7):e42073. doi: 10.1371/journal.pone.0042073. Epub 2012 Jul 31.

Efficient amplification of chimeric adenovirus 5/40S vectors carrying the short fiber protein of Ad40 in suspension cell cultures.

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  • 1Center of Animal Biotechnology and Gene Therapy, and Department of Biochemistry and Molecular Biology, Universitat Autònoma de Barcelona, Barcelona, Spain.

Abstract

The human adenovirus 40 (Ad40) is a promising tool for gene therapy of intestinal diseases. Since the production of Ad40 in vitro is extremely inefficient, chimeric Adenovirus 5/40S vectors carrying the Ad40 short fiber on the Ad5 capsid have been developed. However, Ad5/40S productivity is low. We hypothesized that low productivity was a result of inefficient viral entry into producer cells during amplification. To this end, we have developed a production strategy based on using 211B cells (expressing Ad5 fiber) during amplification steps, while Ad5/40S infectivity is further improved by adding polybrene during infections. In addition, the optimal harvesting time was determined by evaluating the Ad5/40S viral cycle. The developed production strategy significantly reduces the number of amplification cycles and duration of the process. Finally, to further facilitate Ad5/40S production, 211B cells were adapted to suspension thus allowing to easily upscale the production process in bioreactors.

PMID:
22860056
[PubMed - indexed for MEDLINE]
PMCID:
PMC3409147
Free PMC Article

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