Mass Spectrometry Approaches to Glycomic and Glycoproteomic Analyses

Abstract

Glycomic and glycoproteomic analyses involve the characterization of oligosaccharides (glycans) conjugated to proteins. Glycans are produced through a complicated nontemplate driven process involving the competition of enzymes that extend the nascent chain. The large diversity of structures, the variations in polarity of the individual saccharide residues, and the poor ionization efficiencies of glycans all conspire to make the analysis arguably much more difficult than any other biopolymer. Furthermore, the large number of glycoforms associated with a specific protein site makes it more difficult to characterize than any post-translational modification. Nonetheless, there have been significant progress, and advanced separation and mass spectrometry methods have been at its center and the main reason for the progress. While glycomic and glycoproteomic analyses are still typically available only through highly specialized laboratories, new software and workflow is making it more accessible. This review focuses on the role of mass spectrometry and separation methods in advancing glycomic and glycoproteomic analyses. It describes the current state of the field and progress toward making it more available to the larger scientific community.

Global and site‐specific analysis of protein glycosylation in complex biological systems with Mass Spectrometry

“…The applications of glycoproteomics in clinical samples are far more than what we described in this section, there are many comprehensive reviews covering the importance of glycoproteomic technologies in biomedical research (Kim and Varki, ; Tian and Zhang, ; Plomp et al, ; Ruhaak et al, ; Zhang et al, ). Due to the irreplaceable roles of glycoproteins playing in various diseases, high‐throughput glycoproteomic technologies will continue to be developed to enable large‐scale profiling of glycopeptides and glycan structures.…”

Advances in mass spectrometry‐based glycomics

“…N‐ glycans can be efficiently released using several enzymes, such as peptide N‐ glycosidase F (PNGase F), peptide N‐ glycosidase A (PNGase A) and endo ‐β‐ N‐ acetylglucosaminidase H ( Endo H) . Although the most commonly used PNGase F enzyme is not capable of releasing core α1,3 fucosylated glycans, PNGase A can be used instead to achieve the universal release of N‐ glycans.…”

Antibody Panel Based N‐Glycan Imaging for N‐Glycoprotein Biomarker Discovery

“…The discovery of N ‐glycan biomarkers in serum could lead to clinical implementation for disease detection. The majority of serum N ‐glycan methods focus on the analysis of a pool of N ‐glycans released from all proteins in the serum (Gornik et al., ; Kirmiz et al., ; Novokmet et al., ; Reiding et al., ; Ruhaak et al., ; Ruhaak, Xu, Li, Goonatilleke, & Lebrilla, ), or the analysis of one target protein's N ‐glycan profile (Comunale et al., ; Pompach et al., ; Ruhaak et al., , ; Shubhakar et al., ; Simunovic et al., ; Šimurina et al., ; Theodoratou et al., ; Zhang et al., ). Pooled serum analyses have shown trends in overall N ‐glycan changes in the presence of cancer, such as increased fucosylation, branching, and bisects (Gebrehiwot et al., ; Hecht et al., ; Snyder et al., ; Vučković et al., ).…”