518. production of recombinant type 2 adeno-associated virus in bioreactors

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DOIResolve DOI: http://doi.org/10.1016/j.ymthe.2005.07.058
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TypeArticle
Journal titleMolecular Therapy
ISSN1525-0016
1525-0024
Volume11
IssueSupplement 1
PagesS200S201
AbstractRecombinant adeno-associated viruses (rAAV) represent a promising gene therapy vector that may be used in the treatment of diverse human diseases. The major obstacle to broaden their usage is the availability of a production process able to provide sufficient quantities for preclinical and human trials. We present here a successful process for rAAV-2 production in low-serum and serum-free medium performed in a 3L stirred-tank bioreactor. The process is based on the triple transfection of suspension-growing HEK293 cells employing polyethylenimine (PEI) as the DNA carrier. Production of AAV-GFP in a 3L serum-free medium bioreactor yielded titers of 5.1 x 1011 infectious viral particles (IVP), corresponding to 2 x 1012 viral genome (VG) or 6.8 x 1012 viral particles (VP). The cell-specific and total viral titers obtained in suspension culture were about three-fold higher to those obtained with adherent cells. The process is very simple and robust as it does not require a medium exchange, either before or after transfection, making it particularly attractive for the generation of large and homogeneous stocks of rAAV vectors.
Publication date
PublisherElsevier
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedYes
NPARC number23001981
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Record identifier8786f6cb-d0bd-42a4-b52c-3a4ac0e9808f
Record created2017-07-12
Record modified2017-07-12
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