DOI | Resolve DOI: https://doi.org/10.1016/j.ymthe.2005.07.058 |
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Author | Search for: Durocher, Yves1; Search for: Pham, Phuong Lan1; Search for: St-Laurent, Gilles1; Search for: Jacob, Danielle1; Search for: Cass, Brian1; Search for: Nalbantoglu, Josephine; 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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Abstract | Recombinant 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. |
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Publication date | 2005-05 |
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Publisher | Elsevier |
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In | |
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Language | English |
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Peer reviewed | Yes |
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NPARC number | 23001981 |
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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 | 8786f6cb-d0bd-42a4-b52c-3a4ac0e9808f |
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Record created | 2017-07-12 |
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Record modified | 2020-04-07 |
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