Brument Nicole, Morenweiser Robert, Blouin Véronique, Toublanc Estelle, Raimbaud Isabelle, Chérel Yan, Folliot Sébastien, Gaden Florence, Boulanger Pierre, Kroner-Lux Gabrielle, Moullier Philippe, Rolling Fabienne, Salvetti Anna
INSERM ERM 0105 Laboratoire de Thérapie Génique, CHU Hotel Dieu, Nantes, France.
Mol Ther. 2002 Nov;6(5):678-86. doi: 10.1006/mthe.2002.0719.
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.
在此,我们描述了一种基于离子交换树脂的两步色谱法用于纯化重组腺相关病毒2型和5型(rAAV)的方法。体外和体内实验结果表明,该方法无需对细胞裂解物进行任何预纯化步骤,可用于获得高纯度的rAAV2和rAAV5病毒原液。因此,该方法可轻松转移至载体核心实验室并进行放大,从而能够在大型动物模型中直接比较这两种以及其他潜在的AAV血清型。