Production of lentiviral vectors.
Production of lentiviral vectors.
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DOI：
10.1038/mtm.2016.17
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Bovolenta C
中科院分区：
文献类型：
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作者：
Merten OW;Hebben M;Bovolenta C
慢病毒载体（LV）作为基因治疗载体用于治疗获得性和遗传性疾病的使用已经显著增加。本文介绍了这些载体生产的最新技术水平，特别强调了其用于临床目的的大规模生产。与使用稳定生产细胞系生产的肿瘤逆转录病毒载体相反，临床级LV在大多数情况下通过瞬时转染在细胞工厂中生长的293或293T细胞产生。然而，最近的发展也倾向于使用中空纤维反应器、悬浮培养方法和稳定生产细胞系的实施。按照生物技术工业的惯例，已经建立了相当复杂的下游处理方案来去除任何不期望的工艺来源的污染物，例如质粒或宿主细胞DNA或宿主细胞蛋白质。本文比较了已发表的LV的大规模生产和纯化工艺，并介绍了它们的工艺性能。此外，将介绍稳定细胞系领域的发展及其用于临床材料生产载体的方法。
Lentiviral vectors (LV) have seen considerably increase in use as gene therapy vectors for the treatment of acquired and inherited diseases. This review presents the state of the art of the production of these vectors with particular emphasis on their large-scale production for clinical purposes. In contrast to oncoretroviral vectors, which are produced using stable producer cell lines, clinical-grade LV are in most of the cases produced by transient transfection of 293 or 293T cells grown in cell factories. However, more recent developments, also, tend to use hollow fiber reactor, suspension culture processes, and the implementation of stable producer cell lines. As is customary for the biotech industry, rather sophisticated downstream processing protocols have been established to remove any undesirable process-derived contaminant, such as plasmid or host cell DNA or host cell proteins. This review compares published large-scale production and purification processes of LV and presents their process performances. Furthermore, developments in the domain of stable cell lines and their way to the use of production vehicles of clinical material will be presented.