An advanced integrative mass spectrometer - the essential tool for in-depth analysis of diverse biological research.

Lead Research Organisation: University of St Andrews
Department Name: Biology

Abstract

Mass spectrometry is an analysis technique that measures the molecular weight of a molecule, and gives information about its elemental composition. This technique became applicable to biomolecules by the introduction of soft ionisation techniques. Here the intact molecule can be measured and then subsequently fragmented into its component parts. The molecular weight of those parts can be determined, which gives further information about the molecular structure, and the data collectively allows the full identification of the molecule to help answer important biological questions. The development and application of mass spectrometry in the biological field has expanded rapidly over the last few decades and mass spectrometry is now established as a cornerstone of biological analysis. The growth in applications has stemmed from ever increasing gains in resolution and accuracy. A step-change in resolution and accuracy was enabled by the development of the orbitrap mass analyser, which has become an essential instrument in the modern bioanalytical environment.

The Biomedical Sciences Research Complex is a multidisciplinary centre that carries out internationally recognised science at the interface of biology, chemistry, physics and medicine. The mass spectrometry core facility underpins much of this research. However, the current instrumentation is limited and unfortunately some experiments cannot be conducted due to these limitations. This hinders the research and leaves some scientific questions unanswered. We are therefore applying for a Fusion Lumos Orbitrap mass spectrometer with Vanquish UHPLC.

This high end, versatile mass spectrometer will transform a wide range of BBSRC-funded projects in St Andrews and across the UK. The special configuration of the instrument with 3 mass analysers and multiple ways of fragmenting the molecules, alongside the exceptional resolution, will revolutionalise our ability to analyse biomolecules and have a massive impact on the research that we can support, far beyond the boundaries of what we can currently offer with our instrumentation. Researchers from universities and research institutions from across the UK access the facility, and the facility additionally collaborates with industry supporting commercial research, so the benefits will be far reaching, ultimately strengthening our biotechnology and biological research.

The research in this proposal is diverse and encompasses all areas of the BBSRC remit. The applications of the mass spectrometer, and the numerous ways it can benefit the various research projects are equally diverse and impacts all areas of high-end life science research. The versatility of the technique in analysing, identifying and even quantifying a diverse range of molecules from large biomolecules such as proteins and DNA, to small peptides and metabolites, has seen the demand for mass spectrometry analysis expand rapidly. The information derived from a mass spectrometric analysis can be used to define differences in composition and structure of two states of biomolecules and can lead to novel insights into the molecules' role and function. Understanding these aspects of biomolecules is fundamental to research into disease, the ageing process, food security, nutrition, cancer, pathogens, anti-microbial resistance, fundamental biological processes and synthetic biology; several of which are key strategic areas priorities for the BBSRC.

Technical Summary

Mass spectrometry is in increasing demand as an analytical technique for biomolecules. The invention and subsequent commercialisation of orbitrap mass analysers, along with improvements in computing power and software, has revolutionised the range of analysis that can be achieved and biological questions that can be answered. It is essential that the facility is able to take advantage of these developments to enable analysis that will answer the wide range of biological questions that our collaborators - both here in St Andrews and at other universities and industries across the UK - require answers to.

The Fusion Lumos with Vanquish LC offers a unique hardware combination that provides superior analytical performance. Its Tribrid architecture of quadrupole, iontrap and orbitrap allows maximal flexibility of analysis due to the parallel isolation and detection mechanisms, a truly versatile instrument. The multiple fragmentation techniques and MSn capability and its complementary use are essential for greater analysis of biomolecules particularly of post-translational modifications, particularly glycosylation. Targeted and non targeted quantitation techniques will enable quantitation of both proteins and other biomolecules such as carbohydrates, nucleotides and natural products. These non-peptidic biomolecules will additionally benefit from the wide range of column chemistries available at standard flow, enabling analysis in the most challenging of matrices. The dual standard/microflow capabilities will also ensure that proteomics benefits from high resolution with increased analytical selectivity and reduction of the uncertainty associated with the detection of false positives and will allow for a faster turnaround of analysis.

These technical abilities will transform a wide range of projects underpinning research across the BBSRC remit, including nutrition, the ageing process, antimicrobial resistance, pathogens, viruses and synthetic biology.

Planned Impact

Investment in this instrumentation will have far-reaching impacts in the areas of health & disease, nutrition, agriculture, food security, and industrial biotechnology. The revolution in mass spectrometry techniques that the new instrument will bring to the mass spectrometry core facility at the University of St Andrews will have benefits to a wide range of projects, which in turn will have profound effects on knowledge, the economy and society. The impact can be grouped in several areas:

Furthering knowledge:
The research underpinned by bioanalysis carried out on the new instrumentation is likely to have significant downstream impact on a wide range of research into the structure and function of biomolecules. The fundamental research and discoveries will have implications for health, disease, nutrition, agriculture, and biotechnology. Based on the track record of the applicants the enabled research will be presented at international conferences, published in high impact research journals, and where appropriate filed as patent applications. Furthermore, it will be used to attract further research investment to the University of St Andrews.

Industry:
Bioscience in areas such as agrochemicals, pharmaceuticals and the biotechnology industries is vital to the UK economy. Access to cutting-edge mass spectrometry analysis is crucial for small and medium sized research companies, which do not necessarily have the resources or experience to achieve this analysis independently. The BSRC mass spectrometry facility has close links with many companies (eg AlmacDiscovery, Eurofins, WuxiApptec, Crelux, Evotec, NuHana and Pneumagen) advising and assisting in their mass spectrometry analysis, impacting on their research and ultimately having a strong positive effects on the economy. The new instrument will enable further interactions with current collaborators and to form new collaborations with the biosciences industry.

Training and education:
All researchers accessing the facility and the instrument will benefit from the exposure to the latest technologies and results from cutting-edge instrumentation. Underpinned by lectures in the technical theory, undergraduates will gain skills, insights and understanding into the latest mass spectrometry techniques that will be carried with them into their future careers. Hands on access to the instrument will be encouraged for research staff and students where appropriate.

General public and outreach:
Bioscience research and its impact on society can be seen as a remote concept in the general public. The University of St Andrews has committed to engaging with the public and demystifying the role research plays in modern life. The University of St Andrews public engagement team is based in the BSRC building and help researchers plan, develop and evaluate public engagement activity. Since it is not feasible to transport mass spectrometers, we regularly give tours of the lab, including demonstrations to school children and visitors. This includes applicants to the university who, along with their parents, are engaged in a wide variety of public engagement activities, showcasing the mass spectrometry facility and acknowledging the relevant instrument funding bodies.

Publications

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