A versatile and scalable two-step ion-exchange chromatography process for the purification of recombinant adeno-associated virus serotypes-2 and -5.
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.
Brument
Nicole
N
INSERM ERM 0105 Laboratoire de Thérapie Génique, CHU Hotel Dieu, Nantes, France.
Morenweiser
Robert
R
Blouin
Véronique
V
Toublanc
Estelle
E
Raimbaud
Isabelle
I
Chérel
Yan
Y
Folliot
Sébastien
S
Gaden
Florence
F
Boulanger
Pierre
P
Kroner-Lux
Gabrielle
G
Moullier
Philippe
P
Rolling
Fabienne
F
Salvetti
Anna
A
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002586:Cesium / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D002852:Chromatography, Ion Exchange / Q000379:methods*; D002853:Chromatography, Liquid; D000229:Dependovirus / Q000235:genetics* / Q000302:isolation & purification*; D005821:Genetic Techniques; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010957:Plasmids / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014283:Triiodobenzoic Acids / Q000494:pharmacology
D002712:Chlorides; D014283:Triiodobenzoic Acids; D002586:Cesium; C028019:cesium chloride; C044834:iodixanol