| DOI | Resolve DOI: https://doi.org/10.1002/biot.201500176 |
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| Author | Search for: Emmerling, Verena V.; Search for: Pegel, Antje; Search for: Milian, Ernest G.1; Search for: Venereo-Sanchez, Alina; Search for: Kunz, Marion; Search for: Wegele, Jessica; Search for: Kamen, Amine A.; Search for: Kochanek, Stefan; Search for: Hoerer, Markus |
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| Affiliation | - National Research Council of Canada
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| Format | Text, Article |
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| Subject | Bioreactors; Body fluids; DNA; Gene therapy; Mammals; Manufacture; Productivity; Viruses; Adeno-associated virus; Bioprocess optimization; HEK293T; ICELLis; Manufacturing strategy; Recombinant adeno-associated virus; Stirred tank bioreactors; Volumetric productivity; Vectors |
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| Abstract | Viral vectors used for gene and oncolytic therapy belong to the most promising biological products for future therapeutics. Clinical success of recombinant adeno-associated virus (rAAV) based therapies raises considerable demand for viral vectors, which cannot be met by current manufacturing strategies. Addressing existing bottlenecks, we improved a plasmid system termed rep/cap split packaging and designed a minimal plasmid encoding adenoviral helper function. Plasmid modifications led to a 12-fold increase in rAAV vector titers compared to the widely used pDG standard system. Evaluation of different production approaches revealed superiority of processes based on anchorage- and serum-dependent HEK293T cells, exhibiting about 15-fold higher specific and volumetric productivity compared to well-established suspension cells cultivated in serum-free medium. As for most other viral vectors, classical stirred-tank bioreactor production is thus still not capable of providing drug product of sufficient amount. We show that manufacturing strategies employing classical surface-providing culture systems can be successfully transferred to the new fully-controlled, single-use bioreactor system IntegrityTM iCELLisTM. In summary, we demonstrate substantial bioprocess optimizations leading to more efficient and scalable production processes suggesting a promising way for flexible large-scale rAAV manufacturing. |
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| Publication date | 2015-09-24 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 21277505 |
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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 | 155978b4-5776-493c-9910-871683b2cd98 |
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| Record created | 2016-03-09 |
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| Record modified | 2020-04-22 |
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