Universal Method for the Purification of Recombinant AAV Vectors of Differing Serotypes.
Universal Method for the Purification of Recombinant AAV Vectors of Differing Serotypes.
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DOI：
10.1016/j.omtm.2017.12.004
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发表时间：
2018-06-15
期刊：
影响因子：
--
通讯作者：
O'Riordan CR
中科院分区：
文献类型：
--
作者：
Nass SA;Mattingly MA;Woodcock DA;Burnham BL;Ardinger JA;Osmond SE;Frederick AM;Scaria A;Cheng SH;O'Riordan CR
临床良好生产规范（GMP）级腺相关病毒（AAV）载体的产生需要支持产生高纯度载体并且表现出良好的安全性和功效特征的纯化策略。迄今为止，大多数报道的纯化方案是血清型依赖性的，需要为每种AAV基因治疗产品开发方法。在此，我们描述了与多种AAV血清型的纯化相容的平台纯化方法。该方法产生高纯度的载体制剂，其富集具有完整载体基因组的衣壳，并且使空衣壳的分数含量最小化。双柱纯化方法，亲和和离子交换色谱的组合，与通过瞬时三重转染方法或更可扩展的生产细胞系平台产生的一系列AAV血清型相容。总之，所述的适应性纯化方法可用于生产适合在疾病动物模型中进行临床前测试的多种高质量AAV载体。
The generation of clinical good manufacturing practices (GMP)-grade adeno-associated virus (AAV) vectors requires purification strategies that support the generation of vectors of high purity, and that exhibit a good safety and efficacy profile. To date, most reported purification schemas are serotype dependent, requiring method development for each AAV gene therapy product. Here, we describe a platform purification process that is compatible with the purification of multiple AAV serotypes. The method generates vector preparations of high purity that are enriched for capsids with full vector genomes, and that minimizes the fractional content of empty capsids. The two-column purification method, a combination of affinity and ion exchange chromatographies, is compatible with a range of AAV serotypes generated by either the transient triple transfection method or the more scalable producer cell line platform. In summary, the adaptable purification method described can be used for the production of a variety of high-quality AAV vectors suitable for preclinical testing in animal models of diseases.
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DOI：
10.1038/mt.2009.277
发表时间：
2010-03
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
通讯作者：
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影响因子：
3.1
作者：
Davidoff, AM;Ng, CYC;Nathwani, AC
通讯作者：
Nathwani, AC
影响因子：
4.7
作者：
Wang, Qiang;Lock, Martin;Wilson, James M.
通讯作者：
Wilson, James M.
影响因子：
4.2
作者：
Hauswirth, William W.;Aleman, Tomas S.;Jacobson, Samuel G.
通讯作者：
Jacobson, Samuel G.
影响因子：
5.4
作者：
Grimm, Dirk;Lee, Joyce S.;Kay, Mark A.
通讯作者：
Kay, Mark A.