Document Detail


Development of a suspension serum-free helper-dependent adenovirus production system and assessment of co-infection conditions.
MedLine Citation:
PMID:  18079009     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
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:
Type:  Journal Article    
Journal Detail:
Title:  Journal of virological methods     Volume:  148     ISSN:  0166-0934     ISO Abbreviation:  J. Virol. Methods     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-02-22     Completed Date:  2008-05-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8005839     Medline TA:  J Virol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  106-14     Citation Subset:  IM    
Affiliation:
Animal Cell Technology Group, Biotechnology Research Institute, National Research Council, 6100 Royalmount Avenue, Montreal, Quebec, Canada H4P 2R2.
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MeSH Terms
Descriptor/Qualifier:
Adenoviridae / 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:
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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