High Throughput Triage of Enzymatic libraries using membrane and micropatterning

Project ID: 2228cd1262 (You will need this ID for your application)

Under Offer

Research Theme: Manufacturing The Future

UCL Lead department: Biochemical Engineering

Project Summary:

Industrial biocatalysts are essential components in the global push towards “Green Chemistry” methodologies. A well-known challenge in the pipeline of identifying new enzymes is the ability to screen vast landscapes of substrate reactivity/selectivity, reaction conditions, stabilities, and ease of expression for equally large libraries of enzymatic sequences. Cell-free protein synthesis (CFPS) lends itself to the small-scale expression of enzymes by reducing the time taken, simplifying the work up and reducing costs compared to classical E. coli outgrowth on the 96-well scale. This makes CFPS at the small scale a tractable route towards rapidly and repeatedly screening thousands to 10’s of thousands of enzymes, applicable to large scale (meta)genomic libraries as well as mutagenesis libraries. A limit to this approach is the ability to vary multiple reaction conditions e.g. pH, substrate, organic solvent in a single plate for a whole library. This project aims to reconfigure the format of classical plate-based approaches with reagents in microwells, into porous matrix (paper supports) environments. These will act to localise reactions by patterning using pico/nanolitre droplets of expressed proteins to produce arrays for subsequent testing. Methods to localise enzymes, and assess expression, reactivity, selectivity and optimal reaction conditions will be developed. This will enable high throughput techniques to be envisaged to rapidly screen enzymatic libraries for function but could also enable high throughput reaction parameter sweeps, for large scale enzyme libraries. The question here is how high throughput can we go, and how do we do it for membranes?

The student will work in the Dept. of Biochemical Engineering & London Centre for Nanotechnology with co-supervisory team comprised of Dr. Jack Jeffries Michael & Dr. Thomas.