The Use of LC-MS Based Methods for Glycomic Analysis.

Abstract

Increasing interest into the function of glycans in disease and pharmaceuticals means that new and improved methods are required for their analysis. This project has aimed to identify and develop key methods for the analysis of glycans and monosaccharides, which will be of major benefit in further research. The workflow for glycan analysis entails the use of an endoglycosidase, namely PNGase F to cleave N-glycans. This is followed by tagging with a fluorescent label, such as, Rapi-Fluor MS or 2-aminobenzamide for detection by fluorescence using ultra-performance liquid chromatography in combination with mass spectrometry. The analysis of monosaccharides has been carried out using anthranilic acid as a fluorescent tag and 1,2-diamino-4,5-methylenedioxybenzene was used in the analysis of sialic acids. The variety of methods investigated provides the opportunity to select an appropriate method for the sample of interest. Methods of sample preparation and analysis have been established and there is also the possibility of further developing methods for monosaccharide quantification. The described methods will be utilised in ongoing projects and further research.

Date of Award	1 Aug 2019
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Sabine Flitsch (Supervisor), Richard Grencis (Supervisor) & Perdita Barran (Supervisor)