Schnödt Maria, Büning Hildegard
1 Center for Molecular Medicine Cologne, University of Cologne , Cologne, Germany .
2 German Center for Infection Research , Bonn-Cologne and Hannover-Braunschweig (partner sites), Germany .
Hum Gene Ther Methods. 2017 Jun;28(3):101-108. doi: 10.1089/hgtb.2016.188. Epub 2017 Apr 17.
Adeno-associated viral (AAV) vectors have emerged as one of the most popular gene transfer systems in both research and clinical gene therapy. As AAV vectors are derived from a stealth, nonpathogenic virus and lack active integrase activity, these vectors are frequently applied for in vivo gene therapy of liver, muscle, and other postmitotic tissues. Although long-term transgene expression from AAV vector episomes is reported from these tissues, the episomal nature of AAV-once regarded as disadvantage-has become an attractive feature for gene-editing approaches targeting proliferating cells. In response to the high demand, AAV vector production is receiving special attention. Besides particle yields and biological activity, the most important concern is improving vector purity. The most difficult task in this regard is removal of defective particles, that is, capsids that are either empty or contain DNA other than the full-length vector genomes. Herein, we characterize and discuss these so-called product-related impurities, methods for their detection, as well as strategies to avoid or reduce their formation.
腺相关病毒（AAV）载体已成为研究和临床基因治疗中最受欢迎的基因转移系统之一。由于AAV载体源自一种隐匿的、非致病性病毒且缺乏活性整合酶活性，这些载体经常被用于肝脏、肌肉和其他终末分化组织的体内基因治疗。尽管从这些组织中报道了AAV附加体的长期转基因表达，但AAV的附加体性质——曾被视为缺点——已成为针对增殖细胞的基因编辑方法的一个有吸引力的特征。为满足高需求，AAV载体生产正受到特别关注。除了颗粒产量和生物学活性外，最重要的关注点是提高载体纯度。在这方面最困难的任务是去除缺陷颗粒，即要么是空的衣壳，要么包含除全长载体基因组之外的DNA的衣壳。在此，我们对这些所谓的与产品相关的杂质进行表征和讨论，介绍其检测方法以及避免或减少其形成的策略。
