High throughput 2D-IR protein screening - new technology for drug design and diagnostics

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Two-dimensional infrared (2D-IR) is an ultrafast laser spectroscopy method that provides a sensitive 'map' of the 3D structures and intermolecular interactions of biomacromolecules, such as proteins and DNA. 2D-IR complements well-established structure-based biophysical tools like crystallography, cryo-EM or NMR by operating in the solution phase at room temperature and by providing access to ultrafast dynamic motions of molecular structures, which influence ligand binding and so are targets for next-generation drug molecules. This means that 2D-IR has potential as a tool for use in drug design or in measuring protein content in complex mixtures such as biofluids (e.g. blood serum). However, a major barrier to the commercial uptake of 2D-IR has existed in the strong absorption of water (H2O) in the infrared region of the spectrum, which obscured important signatures from proteins and nucleic acids. This overlap of signals from solvent and biomolecule necessitated expensive and labour-intensive isotopic replacement of the solvent with D2O before measurements, making 2D-IR non-viable as a commercial tool for the pharmaceutical industry and incapable of measuring biofluids.This proposal builds on world leading STFC-funded research at the Central Laser Facility that enables the laser pulse sequence used to measure 2D-IR to suppress H2O signals for the first time (Chem Sci 10, 6448 (2019); Editors' Pick of the Week and HOT article collections). This means that the signals from proteins can be cleanly measured without solvent substitution; the protein spectroscopy equivalent of locating the needle in the haystack. Our invention, with established IP (PCT/GB2019/051585) means that we can perform 2D-IR spectroscopy measurements of protein structure and dynamics in physiologically relevant (H2O-based) solvents and biofluids, which removes the major obstacle to commercial implementation of 2D-IR and so opens up the power of 2D-IR to the pharmaceutical and biomedical sectors. To realise this commercial potential, we have formed a partnership with global biopharmaceutical company UCB Pharma and the STFC Central Laser Facility to create the world's first high throughput 2D-IR screening facility. By overcoming the next set of challenges facing commercial 2D-IR implementation, such as rapid, accurate sample delivery and user-friendly data visualisation, the facility will progress 2D-IR to higher TRL levels (3-7), ultimately being able to measure a standard 96-well plate of sub-microlitre volume drug-protein samples and deliver analysis-ready spectra in less than two hours at the press of a button.This facility will deliver straightforward access for end-users to the structural and dynamic details of interactions between biomolecule target and drug candidate and bring a vital new perspective to drug design. In parallel with our primary aim of developing a screening platform for drug design, we will develop the technology further, working with other partners, such as SME ClinSpec DX, to advance 2D-IR screening of biofluids, delivering quantitative protein analysis of biofluids for disease diagnosis and healthcare applications.Successful completion of the project will lay the groundwork for a commercially viable fee-per-sample 2D-IR screening service at the CLF and establish the optimum route to market for 2D-IR analysis.