Fast and Sensitive Mass Spectrometry

We propose to purchase a high-end mass spectrometer system that is capable of high data acquisition speeds and high sensitivity. This is to support high-throughput and reproducible label-free proteomic, metabolomic, lipidomic, and glycomic, workflows. Samples will be analysed in minutes rather than hours, enabling large-scale experiments that are not possible using our existing technology. The system will include rapid liquid chromatography front-ends, gas phase filtering/ion concentration (ion mobility) incorporated within the mass spectrometer system, coupled with rapid fragmentation and high resolution detection technology. High-density data is generated (on the scale of milliseconds), in the form of ion mass-to-charge ratios over time, that can then be processed by specialist software to identify individual analystes and read-out their quantity. Applications range from understanding how nutritional or medicinal compounds in plants vary over different varieties or field conditions, to understanding how the protein chemistry of globally important processes like photosynthetic carbon dioxide fixation works and can be optimized, to discovering new bacterial or fungal enzymes that can efficiently process waste materials. The system will be located in an efficiently run core facility environment, with maintenance, method development, and project workflows closely coordinated between investigators and dedicated expert core-funded facility staff.

We propose to purchase a TimsTOF pro 2 mass spectrometer system, with dual trapped ion mobility separation (Tims) and fast time of flight (TOF) capabilities, with the ion transmission path controlled by Bruker's Parallel Accumulation Serial Fragmentation (PASEF) technique. This core configuration allows ions to be selected based on their shape and/or charge characteristics, and with minimal losses, further transmitted for co-ordinated fragmentation and detection. This gives the dual advantages that ions can be rapidly (millesconds) resolved over chromatographic (seconds) time scales, but also maintained as packets that can be co-coordinately transmitted and detected, which maintains high sensitivity and data density. This in turn means analytes present in highly complex mixtures can be separated for identification and quantification over short time scales; in many cases, using liquid chromatography based separations in minutes rather than hours, compared to standard instrumentation and techniques. We will implement PASEF and related techniques, for proteomic, metabolomic, lipidomic, and glycomic workflows. BBSRC-funded PIs working across the breadth of the BBSRC remit and strategic priorities will be supported by the new equipment; their research will be facilitated and significantly enhanced by access to the new technologies and functionalities. Higher throughput will be especially beneficial for screening large sets of immunopurified protein partners, and more generally in allowing lower-cost processing of high sample numbers (for statistical robustness in proteomics and metabolomics). Tims, at higher resolution settings, will also enable isobar and isomer resolution, and shape-based selection, which will be of great benefit for our workflows on glycans and glycoproteins, and for the selection of chemically crosslinked peptides derived from protein proximity and interactomics studies.