Efficient amplification of chimeric adenovirus 5/40s vectors carrying the short fiber protein of ad40 in suspension cell cultures

Marta Miralles, María Mercedes Segura, Meritxell Puig, Assumpció Bosch, Miguel Chillon

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2 Citations (Scopus)

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. © 2012 Miralles et al.
Original languageEnglish
Article numbere42073
JournalPLoS ONE
Volume7
DOIs
Publication statusPublished - 31 Jul 2012

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