A versatile and scalable two-step ion-exchange chromatography process for the purification of recombinant adeno-associated virus serotypes-2 and -5.

N. Brument (1) , R. Morenweiser (2) , V. Blouin (1) , E. Toublanc (1) , I. Raimbaud (1) , Y. Cherel (3) , S. Folliot (1) , F. Gaden (4) , P. Boulanger (4) , G. Kroner-Lux (5) , P. Moullier (1) , F. Rolling (1) , A. Salvetti (1)

1 Vecteurs viraux et transfert des gènes in vivo
2 Amersham Biosciences Europe GmbH
3 PAnTher - Physiopathologie Animale et bioThérapie du muscle et du système nerveux
4 VPV - Virologie et pathogenèse virale
5 Progen Biotechnik GmbH

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.

Mots clés: Animals; Blotting; Western; Cell Line; Cesium/pharmacology; Chlorides/pharmacology; Chromatography; Ion Exchange/*methods; Liquid; Dependovirus/*genetics/*isolation & purification; Genetic Techniques; Hela Cells; Humans; Mice; Inbred BALB C; Microscopy; Electron; Plasmids/metabolism; Rats; Wistar; Triiodobenzoic Acids/pharmacology

Domaines: Virologie

N. Brument, R. Morenweiser, V. Blouin, E. Toublanc, I. Raimbaud, et al.. A versatile and scalable two-step ion-exchange chromatography process for the purification of recombinant adeno-associated virus serotypes-2 and -5.. 2007. ⟨hal-00128279⟩