Ion Chromatography coupled Mass Spectrometry (IC-MS) for Advanced Structural Characterisation of Carbohydrates

Carbohydrates (or sugars) play myriad roles in nature from energy storage and transport to structural components of cells and tissues to key cell signalling and recognition receptors. Despite the importance of complex carbohydrates (also known as glycans) in all areas of biology, the study of glycans has lagged behind that of the other major biomolecules, especially protein and nucleic acids, due to the vast diversity of structures that exist in nature. This diversity has limited analytical techniques for glycan characterisation. Ion exchange chromatography (IC) with electrochemical detection is one of the core techniques used to separate, identify and quantify complex carbohydrate structures as it provides excellent separation and sensitivity and can be used to detect carbohydrates without having to modify them in any way. However this methodology has been restricted in the past due to its reliance on defined sugar standards for identification. This limitation could be circumvented by linking the IC to a mass spectrometer (MS) for advanced sugar identification and characterisation, but until recently this technology was not possible. Recent exciting advances by the bioscience company ThermoFisher Ltd have enabled the direct linking of the IC to the MS to create a rapid and robust pipeline for advanced carbohydrate analysis. The IC-MS would enable quantification and characterisation of a wide range of complex carbohydrates from human, animal, plant and microbial sources and thus has great potential to transform our ability to analyse and understand the vast diversity of glycan structures that exist and decipher their roles in biology.

This proposal seeks funding for an Ion chromatography (IC) system coupled to an Orbitrap Mass spectrometer (MS). The instrument will provide a state-of-the-art platform for the separation, quantification and structural analysis of complex carbohydrates. High-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) is a sensitive, fast and reliable chromatography method for the separation and quantification of complex carbohydrates, but to date has been limited in its ability to identify and structurally characterise unknown molecules due to the inability to link the IC component to high end MS for structural analysis. Recent exciting technological advances by ThermoFisher have enabled the direct coupling of HPAEC-PAD to a Mass Spectrometer and this new technology offers a transformative platform for the separation, accurate detection and identification of complex carbohydrates; specifically those without robust standards available. The IC-MS will provide transformative technologies to answer research questions aligned to the BBSRC delivery plan including 'Advancing the frontiers of bioscience discovery: Understanding the rules of life and Transformative technologies, Tackling strategic challenges: Bioscience for sustainable agriculture and food and Bioscience for an integrated understanding of health.' This instrument and technology would be the first of its kind within the UK and would transform the analytical capabilities of carbohydrate research both for internal users, but also external users based across the UK. Significantly, the instrument provides a complementary method to the state-of-the-art Carbohydrate Microarray technology already established at Newcastle University. Together the IC-MS and Microarray technologies would establish Newcastle University as a national leader for carbohydrate structural analysis.