Ion mobility mass spectrometer for proteomics facility

Mass spectrometry is a powerful analytical method that is used to identify and quantify compounds from a huge variety of materials, for example, identify enzymes engaged with degradation of waste in soil, or to understand how a cell can divide into two. Mass spectrometry can be used on any type of molecule that can carry a positive or negative charge and is stable in the gas phase at high vacuum. In the Proteomics Research Technology Platform at the University of Warwick we use mass spectrometry to study proteins, which we usually break into small pieces called peptides to make high throughput, large-scale analysis feasible. Proteins provide many vital functions in our bodies from making structures like collagen to working as enzymes to digest our food. In this proposal, we request a new type of mass spectrometer that can separate charged molecules (ions) by their 'shape' (really cross-collisional dimension) as well as separating ions by mass and charge, as all mass spectrometers do.

This extra ability, called ion-mobility, would provide us with a very flexible and exciting instrument, ideal for measuring and quantifying peptides from liquid solutions. Ion mobility means that we would be able to distinguish between very similar peptides that have the same mass, but have, for example, a different order of amino acids, like PEPTIDE and PPEETID. This would be a great improvement over our current instrumentation, where we sometimes see evidence that we have accidently fragmented two different peptides at the same time, which makes the result (the scan) more difficult to interpret.

If funded, we would prioritise time for researchers in BBSRC science areas, such as meeting challenges in Food Security, by deciphering how protein signalling helps plants defend themselves against pathogens. Or by understanding fundamental challenges in how life works, for example in discovering the molecular details of how a yeast cell divides into two using a contractile ring.

We have already developed innovative tools to help teach students and explain mass spectrometry to the public. For example, our animation and "ping-pong ball mass spectrometer" are very popular at open days and can be found on youtube. If funded, we would create new tools to demonstrate ion mobility and invite you to visit us on one of our open days in 2021.

The Proteomics Research Technology Platform (RTP) at Warwick was established in 2014 and currently uses two instruments (an Orbitrap Fusion and Quantiva from Thermo Scientific). Areas of specialism are in peptide-based quantitative proteomics and the analysis of post-translational modifications, and in particular phosphorylation. We wish to provide additional capabilities to academics at Warwick University and external customers by incorporating ion mobility separations. Ion mobility spectrometry coupled to mass spectrometry allows access to a unique dimension for characterisation and separation in complex mixtures. The use of ion mobility separation allows separation of precursors of the same m/z and liquid chromatography retention time which otherwise would be indistinguishable. Additionally, by allowing focussing of ions in time improved signal-to-noise ratios can be obtained for all precursors.

We seek support for an innovative ion mobility mass spectrometry (IM-MS) system. The proposal is based on the timsTOF Pro (Bruker) which that utilises a novel arrangement of two trapped ion mobility separation (TIMS) devices working in parallel. This configuration allows trapped ions to be transferred to the mass spectrometer from the second device whilst the first is being filled with ions allowing use of the full duty cycle. Together with high resolution and ppm mass accuracy this instrument will be transformative in the capabilities that it provides.

If successful the instrument purchase would be taken to public tender and all comparable technologies assessed through a full evaluation process.

We collaborate with industry partners through Warwick Scientific Services on a regular basis and participate at Academic-Industry days, both to showcase technology and also as active scientific researchers.

Other impactful outputs form the purchase of a new mass spectrometry system will be;

- Improved and expanded analytical services in proteomics will be offered to our commercial clients through our existing collaborations.
- New outreach demonstrations will be developed and used to communicate with the public and open days and science fairs.
- We will collaborate with mass spectrometry manufactures in specific analytical challenges (such as the annual international American Society for Mass Spectrometry)
- by hosting one day meetings and training workshops (cost covered by sponsoring companies) and through collaborative PhD studentships.
- We will share training, instrument time and job shadowing with our regional Midlands Innovate partners and local schools
- We will continue to improve our communication of science to the public through interactive demonstrations, videos and aminations and attendance at science fairs and open days. Specifically, we will develop at least one new demonstration tool.
- We will offer increase support for young scientists through mentoring, work experience and career development.