Streamlining workflow and automation to accelerate laboratory scale protein production.
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.
Drug Discovery Program, CRUK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, United Kingdom.
Jennifer
J
Konczal
Drug Discovery Program, CRUK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, United Kingdom. Electronic address: c.gray@beatson.gla.ac.uk.
Christopher H
CH
Gray
D057205:Automation, Laboratory / Q000379:methods*; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry*; D006801:Humans; D011994:Recombinant Proteins / Q000302:isolation & purification*; D057188:Workflow
D011994:Recombinant Proteins
Automated protein purification; Higher throughput expression screening; Multidimensional chromatography; Novel vectors and cloning; Parallel purification; Protein conditioning; Strain engineering