| DOI | Resolve DOI: https://doi.org/10.1016/j.jviromet.2007.10.017 |
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| Author | Search for: Meneses-Acosta, Angélica1; Search for: Dormond, Edwige1; Search for: Jacob, Danielle1; Search for: Tom, Roseanne1; Search for: Bernier, Alice1; Search for: Perret, Sylvie1; Search for: St-Laurent, Gilles1; Search for: Durocher, Yves1; Search for: Gilbert, Rénald1; Search for: Kamen, Amine1 |
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| Affiliation | - National Research Council of Canada. NRC Biotechnology Research Institute
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| Format | Text, Article |
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| Subject | biotechnology; cell line; cell; gene; kidney; serum-free medium; adenovirus; helper-dependent adenovirus; gutless adenovirus; bioreactor production; serum-free suspension culture; MOI; gene therapy |
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| Abstract | Helper-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. |
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| Publication date | 2008-03 |
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| In | |
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| Language | English |
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| NRC number | NRCC 47817 |
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| NPARC number | 12400997 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 3062de35-c152-4bc7-8a0f-3feaedcf4e17 |
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| Record created | 2009-10-26 |
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| Record modified | 2020-04-15 |
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