临床级载体产品的复制型慢病毒分析。
Replication-competent lentivirus analysis of clinical grade vector products.
机构信息
Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.
出版信息
Mol Ther. 2011 Mar;19(3):557-66. doi: 10.1038/mt.2010.278. Epub 2010 Dec 21.
Abstract
Lentiviral vectors are now in clinical trials for a variety of inherited and acquired disorders. A challenge for moving any viral vector into the clinic is the ability to screen the vector product for the presence of replication-competent virus. Assay development for replication-competent lentivirus (RCL) is particularly challenging because recombination of vector packaging plasmids and cellular DNA leading to RCL has not been reported with the current viral vector systems. Therefore, the genomic structure of a RCL remains theoretical. In this report, we describe a highly sensitive RCL assay suitable for screening vector product and have screened large-scale vector supernatant, cells used in vector production, and cells transduced with clinical grade vector. We discuss the limitations and challenges of the current assay, and suggest modifications that may improve the suitability of this assay for screening US Food and Drug Administration (US FDA)-licensed products.
摘要
慢病毒载体目前正在进行各种遗传性和获得性疾病的临床试验。将任何病毒载体推向临床的一个挑战是筛选载体产品中是否存在复制型病毒的能力。复制型慢病毒（RCL）检测方法的开发尤其具有挑战性，因为目前的病毒载体系统尚未报道载体包装质粒和细胞 DNA 重组导致 RCL 的情况。因此，RCL 的基因组结构仍然是理论上的。在本报告中，我们描述了一种适用于筛选载体产品的高灵敏度 RCL 检测方法，并筛选了大规模的载体上清液、用于载体生产的细胞以及用临床级载体转导的细胞。我们讨论了当前检测方法的局限性和挑战，并提出了可能改进该检测方法用于筛选美国食品和药物管理局（US FDA）许可产品的适用性的修改建议。
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