Development of a suspension serum-free helper-dependent adenovirus production system and assessment of co-infection conditions

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DOIResolve DOI: http://doi.org/10.1016/j.jviromet.2007.10.017
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TypeArticle
Journal titleJournal of Virological Methods
Volume148
Issue1-2
Pages106114; # of pages: 9
SubjectAnimal; bio; Biotechnology; Cell Line; Cells; Genes; Human; Kidney; serum-free medium; Adenovirus; Helper-dependent adenovirus; Gutless adenovirus; Bioreactor production; Serum-free suspension culture; MOI; Gene therapy
AbstractHelper-dependent adenovirus (HDAd), deleted in all viral protein-coding sequences has been designed to reduce immune response and favor longterm expression of therapeutic genes in clinical programs. Its production requires co-infection of E1-complementing cells with helper adenovirus (HAd). Significant progresses have been made in the molecular design of HDAd, but large scale production remains a challenge. In this work, a scalable system for HDAd production is designed and evaluated focusing on the co-infection step. A human embryo kidney 293 (293) derived cell line, the 293SF/FLPe was generated to produce efficiently HDAd while restricting the packaging of HAd. This cell line was adapted to grow in suspension and in serum-free medium. Multiplicity of infection (MOI) of HDAd ranging from 0.1 to 50 was evaluated in presence of HAd at a MOI of 5. Optimal MOIs for HDAd amplification were found in the range of 5–10. HAd contamination was only 1%. These results were validated in a 3 L bioreactor under controlled operating conditions where a higher HDAd yield of 2.6×10⁹ viral particles (VP)/mL or 3.5×10⁸ infectious units (IU)/mL of HDAd was obtained.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedNo
NRC number47817
NPARC number12400997
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Record identifier3062de35-c152-4bc7-8a0f-3feaedcf4e17
Record created2009-10-26
Record modified2016-05-09
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