Konczal Jennifer, Gray Christopher H
Drug Discovery Program, CRUK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, United Kingdom.
Drug Discovery Program, CRUK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, United Kingdom.
Protein Expr Purif. 2017 May;133:160-169. doi: 10.1016/j.pep.2017.03.016. Epub 2017 Mar 19.
Protein production facilities are often required to produce diverse arrays of proteins for demanding methodologies including crystallography, NMR, ITC and other reagent intensive techniques. It is common for these teams to find themselves a bottleneck in the pipeline of ambitious projects. This pressure to deliver has resulted in the evolution of many novel methods to increase capacity and throughput at all stages in the pipeline for generation of recombinant proteins. This review aims to describe current and emerging options to accelerate the success of protein production in Escherichia coli. We emphasize technologies that have been evaluated and implemented in our laboratory, including innovative molecular biology and expression vectors, small-scale expression screening strategies and the automation of parallel and multidimensional chromatography.
蛋白质生产设施通常需要为包括晶体学、核磁共振、等温滴定量热法及其他试剂密集型技术等严格的方法生产各种蛋白质。这些团队往往会发现自己成为雄心勃勃的项目流程中的一个瓶颈。这种交付压力促使许多新方法不断发展,以提高重组蛋白生成流程各阶段的产能和通量。本综述旨在描述当前及新兴的加速大肠杆菌中蛋白质生产成功的方法。我们着重介绍已在我们实验室评估和实施的技术,包括创新的分子生物学和表达载体、小规模表达筛选策略以及平行和多维色谱的自动化。