High-throughput analysis of glycosylation

Protein glycosylation - the addition of sugar moieties - plays an important role in determining the stability of proteins and guiding them to specific subcellular compartments. Alterations in glycan structure can modulate the activity of proteins and are strongly linked to diseases such as cancer, diabetes and cardiovascular disorders. In addition, glycoproteins can serve as useful clinical biomarkers in diagnostics as well as targets for disease therapy.

To address the increasing medical need for the characterisation of protein glycosylation, the EU-funded HIGHGLYCAN (Methods for high-throughput (HTP) analysis of protein glycosylation) project is in the process of developing analytical methods for reliably quantifying a multitude of glycan structures in complex biological samples. In this context, they will use ultrahigh-performance liquid chromatography, capillary gel electrophoresis with laser-induced fluorescence detection, and mass spectrometry.

During the first part of the project, researchers focused on the development of suitable sample preparation methods and techniques for the analysis of protein glycosylation. Several improvements were made in measurement techniques and very good progress was achieved with regard to robotisation and automation of glycomic sample preparation methods. This has culminated in several automated profiling workflows of protein glycosylation, which were validated through the analysis of 1 800 samples. Results were further genetically associated with specific single nucleotide polymorphisms, unambiguously establishing the potential of all these methods in high-throughput biomedical research.

Central to the HIGHGLYCAN study is ensuring that the developed technology gets implemented in biotechnological applications. So far, some of the techniques have been incorporated into industrial glycoanalytical services and production of glycoanalytical kits.

Given the structural complexity of glycosylation and the technology gap, the HIGHGLYCAN glycoprofiling system is a reliable solution for the high-throughput identification and quantitation of glycans. Applications include studying glycosylation changes in diseases such as diabetes and identifying diagnostic biomarkers.