Advancing 'omics analysis with a Sciex ZenoToF 7600 mass spectrometer

Understanding the complexity of biology, and engineering biology for our benefit, requires the analysis of a wide range of biomolecules with high accuracy and precision. Mass spectrometry is one of the few techniques capable of providing an insight into this biological complexity, and the data we can generate with mass spectrometry can be used to increase our understanding of how biological systems work. However, the range of molecules important in biology is bewildering. They can be very small, for example the metabolites involved in the conversion of glucose to energy in the cell, to very large, for example the enzymes that catalyse the chemical reactions that make all the other biomolecules. They can also be very stable, such as the collagen that provides the support for the cells in our body or some of the components that make up fats, or very fragile, such as many of the structures added to proteins to control their activity or localization in the cell.

This project is to provide a new generation mass spectrometer that will allow us to perform the complex analysis required to confidently identify the structures of, and measure the amount of many of the molecules that are present in complex biological samples. This is based on improvements in the way that molecules are fragmented in the mass spectrometer to provide information on their structure, enable discrimination between molecules that are only subtly different in structure and provide accurate quantification. The equipment is the first commercial time of flight mass spectrometer that has electron activated dissociation (EAD), a way of fragmenting molecules that is very fast and so gives much more information on structure than other methods, and is able to vary the energy of this fragmentation. Low energy EAD is especially valuable for molecules which have labile chemical groups, as the speed of fragmentation ensures it is not just these that are seen, and high energy EAD allows fragmentation of very stable molecules. It also has improvements that allows small to large fragments to be measured sensitively at the same time, which is critical for large biomolecules and for fast and accurate quantification of specific molecules in complex mixtures.

This instrument will provide benefit to many research programs at the University of Manchester (UoM) in important strategic areas for the country, and in collaboration with a number of major pharmaceutical and biotechnology companies. This includes substantial research in biotechnology, for example in biofuels production and bio-feedstock utilization, and in the development of clean biological catalysts for sustainable chemical production. It also benefits bioscience for health, for example in understanding how the utilization of fats and lipids changes in Parkinson's disease and how inflammation adversely affects biomolecules, and bioscience for sustainable agriculture and food, for example in identifying and understanding the generation of food-based allergens during food processing. The new instrument will generate world-class and underpinning bioscience, for example in the development of analytical technologies and methodologies to understand better the way enzymes work and how biological systems function, and engineering biological processes to use microorganisms to make new chemicals.

Separation and ionization methods coupled to mass spectrometry for the analysis of biomolecules are rapidly improving in development and design, but substantial further progress is needed to identify and quantify the full range of biomolecular species across a range of masses and physiochemistry in a robust manner at medium to high throughput.

This proposal is for a new generation QToF mass spectrometer with significantly improved speed, sensitivity and quality of thermodynamic (collisionally induced) fragmentation, and novel tuneable energy kinetic (electron activated) fragmentation. Current equipment struggles to provide the comprehensive structural identification and quantification across the range of small to large, and labile to highly stable molecules, in complex samples, necessary for understanding and engineering biology. The advances in the SCIEX Zeno TOF 7600 ion optics, provides high speed coupled to sensitivity across the full mass range in fragmentation spectra, and electron activated decomposition, allowing non-ergodic fragmentation of structurally robust singly charged species to highly labile multiply charged biomolecules. This provides a step change in capabilities for robust, sustainable, easy to use mass spectrometers. The instrument will be integrated into the Michael Barber Centre for Collaborative Mass Spectrometry to ensure best use of the instrument's capabilities and sustainability of operation.

This instrument will underpin and enable research in Industrial Biotechnology and Bioenergy, for example in biofuels production and bio-feedstock utilization, Bioscience for Health, for example in understanding role of lipid metabolism in Parkinson's, Bioscience for Sustainable Agriculture and Food in identifying and understanding food-based allergens, and World-class and Underpinning Bioscience, for example in the development of analytical technologies and methodologies, understanding of enzyme mechanisms and systems and syntheti