Advances in mass spectrometry‐based glycomics

Abstract

The diversification of the chemical properties and biological functions of proteins is attained through posttranslational modifications, such as glycosylation. Glycans, which are covalently attached to proteins, play a vital role in cell activities. The microheterogeneity and complexity of glycan structures associated with proteins make comprehensive glycomic analysis challenging. However, recent advancements in mass spectrometry (MS), separation techniques, and sample preparation methods have primarily facilitated structural elucidation and quantitation of glycans. This review focuses on describing recent advances in MS-based techniques used for glycomic analysis (2012-2018), including ionization, tandem MS, and separation techniques coupled with MS. Progress in glycomics workflow involving glycan release, purification, derivatization, and separation will also be highlighted here. Additionally, the recent development of quantitative glycomics through comparative and multiplex approaches will also be described.

Structures and Metabolic Properties of Bovine Milk Oligosaccharides and Their Potential in the Development of Novel Therapeutics

“…Some examples include reducing-end derivatization of glycan sample pairs with a heavy/light label pair (23, 24) or with a series of isobaric reagents (Figure 1B) (25, 26). Further details of these relative quantification techniques and derivatization strategies have been described in a recent book chapter by Orlando (27) and in a review by Dong et al (28), Robinson (29).…”

Characterization of glycan isomers using magnetic carbon nanoparticles as a MALDI co-matrix

“…The utility of MCNPs was then investigated in the glycomic study of N -glycans derived from human blood serum. Human blood serum contains proteins with sophisticated glycosylation, and its profiling plays an essential role in biomarker discovery, and disease diagnosis and prognosis 7,16,18,80,81. Isomers of reduced glycans were first separated by a PGC-nanoLC system and then analyzed with MALDI using MCNPs as a co-matrix.…”

“…Among the variety of analytical methods that have been reported for glycan analysis, mass spectrometry (MS) is considered one of the most prominent methods for glycan profiling 18,19. When coupled with different separation techniques, including ion mobility mass spectrometry,20–23 capillary electrophoresis,24–26 porous graphitized carbon chromatography (PGC),17,27–32 hydrophilic liquid interaction chromatography (HILIC),33,34 and reverse phase chromatography,35 MS is a robust tool for the analysis of isomeric glycan.…”

Glycomics and glycoproteomics: Approaches to address isomeric separation of glycans and glycopeptides

“…The most successful liquid‐phase approaches for the isomeric separation of glycans and glycopeptides have been achieved on nano‐LC systems with porous graphitic carbon (PGC) and alkyl‐bonded (C18) stationary phases using high temperatures or through HILIC with amide embedded groups [3,11]. An additional analytical strategy used in combination with the aforementioned methods is the derivatization of glycans and glycopeptides, in which their physicochemical characteristics are modified to increase the analyte‐stationary phase interactions, enhancing resolution and increasing the analyte ionization efficiency by promoting their detectability [2,12]. The most common derivatization techniques that can be applied for almost all types of isomeric glycan structures are reducing end labeling and permethylation [11,13,14].…”