Voss Carsten
Fermentation Engineering, Faculty of Technology, Bielefeld University, Germany.
Biotechnol Annu Rev. 2007;13:201-22. doi: 10.1016/S1387-2656(07)13008-8.
The concept of curing diseases at the genetic level was already introduced in the 1970s, but only the evolution of molecular biology and tools for genetic manipulation brought the idea into labs and clinics during the last 16 years. Viral and non-viral vectors and delivery systems were developed to transfer therapeutic genes into the target cells. In the case of non-viral approaches plasmid DNA has become a very promising gene delivery vector because it can easily be genetically manipulated and produced by cultivation of plasmid harbouring Escherichia coli and subsequent downstream processing, thus making production easy in comparison to other gene delivery vectors. Another advantage in using plasmid DNA is the low risk of immunogenic reactions and oncogen activation that can arise while using viral vectors. This review describes the recent development in plasmid manufacturing ranging from bacterial cultivation in batch and fedbatch mode to produce plasmid-bearing E. coli over cell lysis and subsequent purification to storage, application, and process and quality control.
在基因水平上治疗疾病的概念早在20世纪70年代就已提出，但直到过去16年，分子生物学和基因操作工具的发展才使这一想法进入实验室和临床。人们开发了病毒和非病毒载体及递送系统，将治疗性基因导入靶细胞。在非病毒方法中，质粒DNA已成为一种非常有前景的基因递送载体，因为它可以很容易地进行基因操作，并通过培养携带质粒的大肠杆菌及后续下游加工来生产，因此与其他基因递送载体相比，生产更容易。使用质粒DNA的另一个优点是，与使用病毒载体时可能出现的免疫原性反应和致癌基因激活风险较低。本综述描述了质粒生产的最新进展，从分批和补料分批模式培养细菌以生产携带质粒的大肠杆菌，到细胞裂解及随后的纯化、储存、应用以及过程和质量控制。
