| DOI | Trouver le DOI : https://doi.org/10.1002/biot.201500176 |
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| Auteur | Rechercher : Emmerling, Verena V.; Rechercher : Pegel, Antje; Rechercher : Milian, Ernest G.1; Rechercher : Venereo-Sanchez, Alina; Rechercher : Kunz, Marion; Rechercher : Wegele, Jessica; Rechercher : Kamen, Amine A.; Rechercher : Kochanek, Stefan; Rechercher : Hoerer, Markus |
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| Affiliation | - Conseil national de recherches du Canada
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| Format | Texte, Article |
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| Sujet | 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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| Résumé | 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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| Date de publication | 2015-09-24 |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Numéro NPARC | 21277505 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | 155978b4-5776-493c-9910-871683b2cd98 |
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| Enregistrement créé | 2016-03-09 |
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| Enregistrement modifié | 2020-04-22 |
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