Nass Shelley A, Mattingly Maryellen A, Woodcock Denise A, Burnham Brenda L, Ardinger Jeffrey A, Osmond Shayla E, Frederick Amy M, Scaria Abraham, Cheng Seng H, O'Riordan Catherine R
Gene Therapy, Sanofi, 49 New York Avenue, Framingham, MA 01701, USA.
Mol Ther Methods Clin Dev. 2017 Dec 22;9:33-46. doi: 10.1016/j.omtm.2017.12.004. eCollection 2018 Jun 15.
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.
临床药品生产质量管理规范（GMP）级腺相关病毒（AAV）载体的生产需要能够支持高纯度载体生产且具有良好安全性和有效性的纯化策略。迄今为止，大多数报道的纯化方案都依赖于血清型，每种AAV基因治疗产品都需要进行方法开发。在此，我们描述了一种适用于多种AAV血清型纯化的平台纯化工艺。该方法可生成高纯度的载体制剂，这些制剂富含具有完整载体基因组的衣壳，并将空衣壳的比例降至最低。这种双柱纯化方法是亲和色谱和离子交换色谱的组合，适用于通过瞬时三重转染法或更具扩展性的生产细胞系平台产生的一系列AAV血清型。总之，所描述的这种适应性强的纯化方法可用于生产各种适用于疾病动物模型临床前测试的高质量AAV载体。
