Public Funding for Douglas Instruments Limited

In roughly the last five years it has become possible to determine the structures of macromolecules such as protein complexes and membrane proteins at near-atomic resolution using single particle cryogenic electron microscopy (cryoEM). These developments have increased the need to explore chemical space in order to get samples ready for cryoEM. Biophysical methods such as dynamic light scattering (DLS) and thermal shift assays can be used for this exploration. The company is now seeking to market its robotics -- currently used as automated crystallization drop-setters -- as dispensers for high throughput biophysical analysis of samples for cryoEM, using new software. However, recommendations for the exploration of chemical space will also be required, and these are, so far, lacking. This A4I proposal aims to help by listing one or more sets of solutions that can act as "screens". A screen will comprise a set of (e.g., 48) chemical solutions that have given favourable results in the past. In use, each solution would be mixed with a new sample, following a constant workflow for all samples. The objective is to identify reagents that alter the behaviour of the target in favourable ways, usually by breaking up aggregation. (This is similar to screening in protein crystallization, but the chemical space explored and the assay used will be different.) The objective is to find suitable starting conditions for protein structure determination. This new work-flow will dramatically reduce the number of trials needed for protein structure determination especially by cryoEM, and increase the throughput of structural biology labs. It will thus open a new market for the company's robotic platforms in the context of sample preparation for single particle cryoEM.