Scalable Production of AAV Vectors in Orbitally Shaken HEK293 Cells.

Détails

ID Serval
serval:BIB_29064F6BCC17
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Scalable Production of AAV Vectors in Orbitally Shaken HEK293 Cells.
Périodique
Molecular therapy. Methods & clinical development
Auteur(s)
Blessing D., Vachey G., Pythoud C., Rey M., Padrun V., Wurm F.M., Schneider B.L., Déglon N.
ISSN
2329-0501 (Print)
ISSN-L
2329-0501
Statut éditorial
Publié
Date de publication
14/06/2019
Peer-reviewed
Oui
Volume
13
Pages
14-26
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: epublish
Résumé
Adeno-associated virus (AAV) vectors are currently among the most commonly applied for <i>in vivo</i> gene therapy approaches. The evaluation of vectors during clinical development requires the production of considerable amounts of highly pure and potent vectors. Here, we set up a scalable process for AAV production, using orbitally shaken bioreactors and a fully characterized suspension-adapted cell line, HEKExpress. We conducted a proof-of-concept production of AAV2/8 and AAV2/9 vectors using HEKExpress cells. Furthermore, we compared the production of AAV2/9 vectors using this suspension cell line to classical protocols based on adherent HEK293 cells to demonstrate bioequivalence <i>in vitro</i> and <i>in vivo</i> . Following upstream processing, we purified vectors via gradient centrifugation and immunoaffinity chromatography. The <i>in vitro</i> characterization revealed differences due to the purification method, as well as the transfection protocol and the corresponding HEK293 cell line. The purification method and cell line used also affected <i>in vivo</i> transduction efficiency after bilateral injection of AAV2/9 vectors expressing a GFP reporter fused with a nuclear localization signal (AAV2/9-CBA-nlsGFP) into the striatum of adult mice. These results show that AAV vectors deriving from suspension HEKExpress cells are bioequivalent and may exhibit higher potency than vectors produced with adherent HEK293 cells.
Mots-clé
AAV8, AAV9, CNS transduction, adeno-associated virus, immune affinity chromatography, orbitally shaken bioreactors, suspension HEK293 cells, transient transfection
Pubmed
Open Access
Oui
Création de la notice
23/01/2019 12:06
Dernière modification de la notice
08/05/2019 16:10
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