A rapid scanning ion mobility MALDI-TOF mass spectrometry platform for state of the art metabolomics, proteomics and molecular imaging

Mass spectrometry (MS) is a critical analytical technology that underpins a significant research portfolio across the School of Biosciences at The University of Sheffield (TUoS). This research involves understanding biochemical pathways at a molecular level through the analysis of proteins and metabolites. The demand for more specialised and increased information from mass spectrometry has led to the need for investment in advanced technologies with enhanced capabilities above our current instrumentation. These limitations include depth of coverage (ability to detect less abundant biomolecules), amount of sample required for analysis, and lack of detailed spatial distribution information. The acquisition of this new instrument will enable a step-change in our ability to undertake sophisticated molecular analysis of complex biological systems aligned to the BBSRC remits of advancing the frontiers of bioscience discovery and tackling strategic challenges1. In particular targeting the priorities of bioscience for sustainable agriculture and food and understanding the rules of life.

The objective of this initiative is to acquire the Bruker timsTOF fleX MALDI-22 mass spectrometer. This cutting-edge instrument provides superior speed and sensitivity compared to our existing equipment which facilitates a substantial boost in sample throughput for analysis. Furthermore, it boasts the capability for mass spectrometry imaging to measure the spatial localisation of metabolites or proteins within a tissue section. This dual functionality of determining both the "what" (identity) and "where" (spatial distribution) of specific metabolites or proteins provides valuable insights into the biochemical and cellular processes underlying biological phenomena.

Existing technological limitations in spatial resolution for imaging mass spectrometry have posed a significant barrier to advancing our understanding of metabolism. The inability to measure the localisation of metabolites at a cellular level has been a major bottleneck. The advances in technology and the introduction of the timsTOF fleX MALDI-2 instrument has made single cell imaging a possibility with the ability to image routinely down to a level of 5µm. This improved resolution opens up a diverse array of research possibilities.

The proposed uses of the instrumentation will serve key strategic BBSRC research priorities by providing advanced analytical technologies to researchers, at all stages of their careers, across TUoS and beyond. For example, a better understanding about biochemical pathways in agriculture allows us to find early biomarkers of disease or develop targeted pesticide or herbicide usage leading to reduced applications of these chemicals. It can also inform us as to the mechanisms of pollinator and pathogen interactions with plants and therefore how this can be managed in a more sustainable way. The advancement in the technology allows us to increase our capabilities in other areas within the BBSRC remit including bioscience for an integrated understanding of health by studying ageing and the immune system using a drosophila (fruit fly) model organism. Securing this equipment is essential for expanding the frontiers of our existing research portfolio within the BBSRC priorities.