Label-free GAG disaccharide analysis by HILIC-MS/MS for studying diverse biological sample types

Abstract The extracellular matrix (ECM) and cell surface glycocalyx are key components of biology and play crucial roles in development and tissue function, as well as disease. Proteoglycans, and their glycosaminoglycan (GAG) side chains, are critical components of the ECM and the glycocalyx. GAGs can bind to many different proteins, such as chemokines, and form hydrated barriers around cells. Existing and new methods are helping us to uncover more about the roles of GAGs in biology. Here, we expand on existing technologies and provide streamlined, standardised and well-documented methods that can be easily adopted in standard analytical facilities. We provide extensive detailed step-by-step guides describing sample disruption, GAG disaccharide preparation from biological tissues and their analysis by HILIC-MS/MS. In addition, we demonstrate utility of this method when using a range of different samples as biological sources. This method will sit alongside existing and new techniques to help improve access to GAG analysis, and thereby further the field of understanding GAG function in complex biological contexts.