Chambers Stephen P, Fulghum John R, Austen Douglas A, Lu Fan, Swalley Susanne E
Gene Expression, Vertex Pharmaceuticals, Cambridge, MA, USA.
Methods Mol Biol. 2009;498:143-56. doi: 10.1007/978-1-59745-196-3_10.
The production of recombinant proteins usually involves the exploration of a wide variety of expression and purification methodologies in the pursuit of a strategy tailored to a particular protein. The methods applied are reliant on exploiting individual differences between expression systems or the variations in specific protein properties. These bespoke strategies have not lent themselves to high-throughput methodologies. Ultimately the development of robust generic methods capable of simplifying and stabilizing the process, allowing automation, was necessary to increase throughput. This chapter describes a series of high-throughput methods used to express, purify, and quantify recombinant protein produced in E. coli or insect cells.
重组蛋白的生产通常需要探索各种各样的表达和纯化方法，以寻求适合特定蛋白的定制策略。所应用的方法依赖于利用表达系统之间的个体差异或特定蛋白质特性的变化。这些定制策略并不适用于高通量方法。最终，开发能够简化和稳定过程、实现自动化的强大通用方法对于提高通量是必要的。本章描述了一系列用于表达、纯化和定量在大肠杆菌或昆虫细胞中产生的重组蛋白的高通量方法。
