Title:
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Efficient amplification of chimeric adenovirus 5/40S vectors carrying the short fiber protein of Ad40 in suspension cell cultures
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Author:
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Miralles, Marta; Segura, Maria Mercedes; Puig Ferrer, Meritxell; Bosch i Merino, Assumpció; Chillón Rodríguez, Miguel
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Abstract:
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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 |
Subject(s):
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-Vector-borne diseases -Viral vectors -Viral tropism -Cell cultures -Viral packaging -Penicillin -Streptomycin -Gene therapy |
Rights:
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open access
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Document type:
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Article |
Published by:
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Share:
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Uri:
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https://ddd.uab.cat/record/128626
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