Lin Chiann-Tso, Moore Priscilla A, Kery Vladimir
Molecular Biosciences, Pacific Northwest National Laboratory, Richland, WA, USA.
Methods Mol Biol. 2009;498:129-41. doi: 10.1007/978-1-59745-196-3_9.
Functional genomics and the application of high-throughput (HT) approaches to solve biological and medical questions are the main drivers behind the increasing need for HT parallel expression and purification of recombinant proteins. Automation is necessary to facilitate this complex multistep process. We describe, in detail, an HT-automated purification of hexahistidine-tagged recombinant proteins using MagneHis Ni-Particles and the Biomek FX robot. This procedure is universally applicable to hexa-histidine-tagged recombinant proteins with the tag positioned at either the N- or C-terminus. With minor modifications, the automated protein purification protocol presented in this chapter could be adapted to purify recombinant proteins bearing other tags than hexahistidine and/or other expression systems than E. coli.
功能基因组学以及应用高通量（HT）方法解决生物学和医学问题是对重组蛋白进行高通量平行表达和纯化需求不断增加的主要驱动因素。自动化对于促进这一复杂的多步骤过程是必要的。我们详细描述了使用MagneHis Ni颗粒和Biomek FX机器人对六组氨酸标签重组蛋白进行高通量自动化纯化。该程序普遍适用于标签位于N端或C端的六组氨酸标签重组蛋白。只需进行少量修改，本章介绍的自动化蛋白纯化方案就可适用于纯化带有除六组氨酸以外其他标签的重组蛋白和/或除大肠杆菌以外的其他表达系统。
Zotero 插件