A versatile and scalable two-step ion-exchange chromatography process for the purification of recombinant adeno-associated virus serotypes 2 and 5.
2006
N Morenweiser R Blouin V Toublanc E Raimbaud I Cherel y Folliot S Gaden F Boulanger P Kroner-Lux G Moullier P Rolling F Salvetti A., Brument | Virologie et pathogenèse virale (VPV) ; Université Claude Bernard Lyon 1 (UCBL) ; Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS) | Laboratoire de Thérapie Génique (1INSERM ERM 0105) ; CHU Hotel Dieu | Munzingerstrasse 9 (AMERSHAM BIOSCIENCES EUROPE GMBH) ; Munzingerstrasse 9 | Physiopathologie Animale et bioThérapie du muscle et du système nerveux (PAnTher) ; École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Progen Biotechnik GmbH (MAASSTRASSE 30PROGEN BIOTECHNIK GMBH) ; Maasstrasse 30 | This work was supported by Amersham Biosciences, the Association Française contre les Myopathies, Vaincre les Maladies Lysosomales, the Association Nantaise de Thérapie Génique, and the Fondation pour la Thérapie Génique en Pays de la Loire. F.G. was a recipient of a doctoral fellowship from the French Association Vaincre la Mucoviscidose.
Here we describe the development of a two-step chromatography process based on the use of ion-exchange resins for the purification of recombinant adeno-associated virus (rAAV) serotypes-2 and-5. In vitro and in vivo results demonstrate that this method, which does not require any prepurification step of the cell lysate, can be applied to obtain highly pure rAAV2 and rAAV5 stocks. As such,this procedure can be easily transferred in vector cores and also scaled up, allowing the direct comparison of these two, and potentially other, AAV serotypes in large animal models.
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