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Case study: Protein production of Test Protein 1

Through our Centre for Automated Protein Research and Innovation (CAPRI), we successfully expressed and purified Test Protein 1 in both E. coli and insect cell systems for Dr Emma Fairweather at Sulantrix Ltd. Using our optimised expression and purification protocols, we delivered large-scale quantities of high-quality protein, enabling downstream structural and functional studies.

Research challenge and facility involvement

Efficient expression and purification of recombinant proteins can be challenging, often requiring tailored strategies for each target. CAPRI actively collaborated with the research team to determine the most suitable conditions for each protein production.

Test Protein 1 was produced at a 4 L culture scale in E. coli and insect cells, with flexible volumes available upon request. Purification strategies included affinity, ion-exchange, and size-exclusion chromatography, performed using the AKTA Pure system.

For Test Protein 1, a two-step process of nickel-affinity chromatography followed by size-exclusion chromatography successfully yielded highly purified protein from both expression systems.

Outcome and impact

The optimised workflow enabled the successful production of Test Protein 1 in two different expression systems.

E. coli system: 138.2 mg of purified protein obtained from 4L culture

Images of SDS-PAGE gels, stained with coomassie blue, highlighting protein bands in dark blue with lighter blue for the gel background. The left-hand side of the image contains a protein ladder marker on the gel, and multiple bands before protein purification, the right-hand side of the image contains fewer bands and only shows the protein of interest which has been purified.

Insect cell system: 1.35 mg of purified protein obtained from 4 L culture

Images of SDS-PAGE gels, stained with coomassie blue, highlighting protein bands in dark blue with lighter blue for the gel background. The left-hand side of the image contains a protein ladder marker on the gel, and multiple bands before protein purification, the right-hand side of the image contains fewer bands and only shows the protein of interest which has been purified.

These results demonstrate the value of CAPRI’s tailored support and highlight the facility’s capacity to accelerate diverse protein production projects.

Quick details

Principal Investigator: Professor Patrick Eyers
Shared Research Facility used: Centre for Automated Protein Research and Innovation (CAPRI)
Instruments: AKTA Pure
Contact for facility access: capri@liverpool.ac.uk