High-Throughput Expression Screening and Purification of Recombinant Proteins in E. coli

Abstract

The protocols outlined in this chapter allow for the small-scale test expression of a single or multiple proteins concurrently using several expression conditions to identify optimal strategies for producing soluble, stable proteins. The protocols can be performed manually without the need for specialized equipment, or can be translated to robotic platforms. The high-throughput protocols begin with transformation in a 96-well format, followed by small-scale test expression using auto-induction medium in a 24-well format, finishing with purification in a 96-well format. Even from such a small scale, there is the potential to use the purified proteins for characterization in pilot studies, for sensitive micro-assays, or for the quick detection of and differentiation of the expected size and oxidation state of the protein by mass spectrometry.