We develop and use efficient parallel DNA cloning and protein production systems using bacterial or eukaryotic cells for making the proteins.
The Protein Production Small Research Facility (SRF) sits within the Centre for Medicines Discovery (CMD) and provides a high-quality protein production service for both academics and industrial scientists. Our research is based on 16 years of experience in protein production in the Structural Genomics Consortium (SGC), where we used cutting-edge technologies and high-throughput (HTP) platforms to express a plethora of very challenging human proteins for structural and functional analyses. These include integral membrane proteins, intrinsically disordered proteins, and extracellular secreted proteins. We have devised methodologies for HTP cloning in 96-well format to increase the speed of generation of constructs for testing, and miniaturised and streamlined expression platforms for E. coli, insect and mammalian cell cultures for both soluble and membrane proteins. These technologies, tailored by my team enabled the SGC in Oxford to solve more than 2000 human protein structures and fulfil the goals of our awarded grants.
In August 2020, our Protein Production SRF was set up to support internal protein requirements within Oxford, external academic research and industry needs. The range of activities that we offer is provided below. Quality control by intact mass analysis is done on every protein generated via our internal MS SRF (see below).
Protein Production activities offered:
- Generation of high-throughput cloning and expression screening protocols enabling screening of >100,000 96-well plates of constructs in 16 years [1-6].
- Establishment of protocols and protein engineering to produce an array of epigenetic proteins including bromodomains and lysine demethylases [7-13].
- Development of expression systems and reproducible protocols to enable screening and production of integral membrane proteins [14-22].
- Creation of a toolkit of vectors for screening and production of proteins and strategies for identifying protein-protein interactions (Published shortly) .
- Production of a highly reproducible Spike antigen for use in a serology assay platform, developed at the University of Oxford, for identifying antibodies against SARS-CoV-2 [24, 25].