title: Efficient amplification of chimeric adenovirus 5/40S vectors carrying the short fiber protein of Ad40 in suspension cell cultures.
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.
authors.affiliation: Center of Animal Biotechnology and Gene Therapy, and Department of Biochemistry and Molecular Biology, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Miralles
authors.forename: María Mercedes
authors.initials: MM
authors.lastname: Segura
authors.forename: Meritxell
authors.initials: M
authors.lastname: Puig
authors.forename: Assumpció
authors.initials: A
authors.lastname: Bosch
authors.forename: Miguel
authors.initials: M
authors.lastname: Chillon
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D002678:Chimera; D005822:Genetic Vectors; D006801:Humans; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D014764:Viral Proteins