Koehn Jim, Hunt Ian
Protein Structure Unit, Centre for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
Methods Mol Biol. 2009;498:1-18. doi: 10.1007/978-1-59745-196-3_1.
Recombinant protein production plays a crucial role in the drug discovery process, contributing to several key stages of the pathway. These include exploratory research, target validation, high-throughput screening (HTS), selectivity screens, and structural biology studies. Therefore the quick and rapid production of high-quality recombinant proteins is a critical component of the successful development of therapeutic small molecule inhibitors. This chapter will therefore attempt to provide an overview of some of the current "best-in-class" cloning, expression, and purification strategies currently available that enhance protein production capabilities and enable greater throughput. As such the chapter should also enable a reader with limited understanding of the high-throughput protein production (HTPP) process with the necessary information to set up and equip a laboratory for multiparallel protein production.
重组蛋白生产在药物发现过程中起着至关重要的作用,对该途径的几个关键阶段都有贡献。这些阶段包括探索性研究、靶点验证、高通量筛选(HTS)、选择性筛选和结构生物学研究。因此,快速生产高质量的重组蛋白是治疗性小分子抑制剂成功开发的关键组成部分。本章将试图概述一些目前可用的“同类最佳”克隆、表达和纯化策略,这些策略可提高蛋白生产能力并实现更高的通量。因此,本章还应能为对高通量蛋白生产(HTPP)过程了解有限的读者提供必要信息,以便建立和装备一个用于多平行蛋白生产的实验室。