High throughput protein analysis in complex matrices using ultrafast 2D-IR spectroscopy

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Background: The continuous development of ultrafast laser technology over the last decade has delivered the ability to acquire information-rich 2D-IR spectroscopy datasets at ever increasing rates. Currently, one 2D-IR spectrum can be acquired in less than a minute using 100 kHz repetition rate laser systems, while the advent of next generation technology at national facilities is expected to decrease this further. However, whilst the efficiency of data collection has increased enormously, the barrier to 2D-IR generating impact in biological and biomedical arenas is the efficient and effective handling of large amounts of spectral data. This challenge is twofold 1) First and foremost is the need to obtain, process and present data at a rate that is comparable to the measurement time (building on our recent work J. Chem. Phys. 157, 205102 (2022)) 2) There is a now a realisation that advanced computational 'big-data' methodologies hold the key to unlocking the complexities of 2D-IR datasets. Solving these two issues will require not only automated sampling systems to record data libraries but also the application of machine Learning or Artificial Intelligence-driven data analysis routines.Another development that we seek to capitalise on is our innovative work that has enabled us to move away from the conventional methods of obtaining 2D-IR spectra in D2O to avoid natural water interference that obscures protein spectral signatures. We have taken a world-lead in the analysis of protein structure and dynamics in native solvents (e.g. Chemical Science, 10, 6448-6456 (2019)). This has led to proof-of-concept applications of 2D-IR for high throughput protein-drug screening and molecular analysis of blood serum samples, including drug binding to the serum albumin protein in blood serum (Analyst, 147, 3464-3469 (2022).ObjectivesBuilding on the recent installation of world-leading UKRI-funded laser technology at the University of York and the development of automated sample that is handling currently underway, the aim is to establish automated data processing, visualisation and high-level analysis methods for 2D-IR spectroscopy. Testing and development phases at York in collaboration with the Central Laser Facility (CLF) will then progress to technology transfer to the CLF for the benefit of the user community. Scientifically, we will focus on the development of biomolecular (protein and nucleic acid) analysis in complex biological matrices (biofluids, soft tissue and bone) in order to meet varying challenges in sample presentation, data acquisition and analysis. Access to the York instrument will confer the ability to acquire large data libraries to support ML/AI data analysis methods. The student will provide a key state of readiness to optimise uptake of the new CLF technology.