Production of adeno-associated virus (AAV) serotypes by transient transfection of HEK293 cell suspension cultures for gene delivery

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DOIResolve DOI: http://doi.org/10.1016/j.jviromet.2013.10.038
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TypeArticle
Journal titleJournal of Virological Methods
ISSN0166-0934
Volume196
Pages163173; # of pages: 11
Subjectadenovirus vector; butyric acid; plasmid DNA; Adeno associated virus; article; bioreactor; controlled study; culture medium; embryo; human; human cell; nonhuman; priority journal; scale up; serotype; suspension cell culture; viral gene delivery system; virogenesis; virus genome; virus infectivity
AbstractAdeno-associated virus (AAV) is being used successfully in gene therapy. Different serotypes of AAV target specific organs and tissues with high efficiency. There exists an increasing demand to manufacture various AAV serotypes in large quantities for pre-clinical and clinical trials. A generic and scalable method has been described in this study to efficiently produce AAV serotypes (AAV1-9) by transfection of a fully characterized cGMP HEK293SF cell line grown in suspension and serum-free medium. First, the production parameters were evaluated using AAV2 as a model serotype. Second, all nine AAV serotypes were produced successfully with yields of 1013Vg/L cell culture. Subsequently, AAV2 and AAV6 serotypes were produced in 3-L controlled bioreactors where productions yielded up to 1013Vg/L similar to the yields obtained in shake-flasks. For example, for AAV2 1013Vg/L cell culture (6.8×1011IVP/L) were measured between 48 and 64h post transfection (hpt). During this period, the average cell specific AAV2 yields of 6800Vg per cell and 460IVP per cell were obtained with a Vg to IVP ratio of less than 20. Successful operations in bioreactors demonstrated the potential for scale-up and industrialization of this generic process for manufacturing AAV serotypes efficiently. © 2013.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); Human Health Therapeutics (HHT-TSH)
Peer reviewedYes
NPARC number21270779
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Record identifierf1fac518-73bf-4f56-a8b0-ff253ef69920
Record created2014-02-17
Record modified2016-05-09
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