| Development of a suspension serum-free helper-dependent adenovirus production system and assessment of co-infection conditions. | |
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MedLine Citation:
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PMID: 18079009 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Helper-dependent adenovirus (HDAd), deleted in all viral protein-coding sequences has been designed to reduce immune response and favor long-term 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 x 10(9) viral particles (VP)/mL or 3.5 x 10(8) infectious units (IU)/mL of HDAd was obtained. |
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Authors:
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Angélica Meneses-Acosta; Edwige Dormond; Danielle Jacob; Roseanne Tom; Alice Bernier; Sylvie Perret; Gilles St-Laurent; Yves Durocher; Rénald Gilbert; Amine Kamen |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of virological methods Volume: 148 ISSN: 0166-0934 ISO Abbreviation: J. Virol. Methods Publication Date: 2008 Mar |
Date Detail:
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Created Date: 2008-02-22 Completed Date: 2008-05-13 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8005839 Medline TA: J Virol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 106-14 Citation Subset: IM |
Affiliation:
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Animal Cell Technology Group, Biotechnology Research Institute, National Research Council, 6100 Royalmount Avenue, Montreal, Quebec, Canada H4P 2R2. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adenoviridae
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growth & development* Adenovirus E1 Proteins / genetics Cell Count Cell Line Cell Survival Culture Media, Serum-Free Genetic Vectors Helper Viruses / physiology Humans Transduction, Genetic Virus Cultivation / methods* |
| Chemical | |
Reg. No./Substance:
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0/Adenovirus E1 Proteins; 0/Culture Media, Serum-Free |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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