Recent advances in the MS analysis of glycoproteins: Capillary and microfluidic workflows

Abstract

Recent developments in bioanalytical instrumentation, mass spectrometry (MS) detection and computational data analysis approaches have provided researchers with capabilities for interrogating the complex cellular glycoproteome, to help gain a better insight into the cellular and physiological processes that are associated with a disease, and to facilitate the efforts centered on identifying disease-specific biomarkers. This review describes the progress achieved in the characterization of protein glycosylation by using advanced capillary and microfluidic-MS technologies. The major steps involved in large-scale glycoproteomic analysis approaches are discussed, with special emphasis given to workflows that have evolved around complex MS detection functions. In addition, quantitative analysis strategies are assessed, and the bioinformatics aspects of glycoproteomic data processing are summarized. The developments in commercial and custom fabricated microfluidic front-end platforms to ESI- and MALDI-MS instrumentation, for addressing major challenges in carbohydrate analysis such as sensitivity, throughput and ability to perform structural characterization, are further evaluated and illustrated with relevant examples.

Nanofluidics-Based Mass Spectrometry. Applications for Biomarker Discovery in Lysosomal Storage Diseases

“…Different configurations such as single and triple quadrupole, TOF, IT, and ultra-high-resolution FTICR mass spectrometers were adapted to chipbased ESI and contributed significant benefits for various studies. However, as nicely illustrated already by a number of reviews [31][32][33] the general areas of implementation and applications of chip technologies either silicon-based, glass, or polymer microchips have by far been primarily proteomics, genomics, cell analysis, drug discovery, glycomics and metabolomics.…”

Development of Monolithic Column Materials for the Separation and Analysis of Glycans

“…Nano-ESI-MS improved the detection limits down to attomole level with use of ion-trap MS (IT-MS) with the direct infusion of a sample solution [64,65]. More developments in MS instruments utilized in glycan analysis are summarized in several reviews [66][67][68][69][70]. Though the benefits of glycan analysis in using MS instruments are overwhelming, there are still some limitations such as non-selective ionization and fragmentation of peptides and degradation of the samples during the sequence of excitation, trapping and detection.…”

Analytical glycobiology at high sensitivity: current approaches and directions

“…While the capillary-based glycan separations of biomedical interest have been amply demonstrated in the literature, there is a general trend toward the use of chip-based separatory systems [212-216], which offer substantial gains in terms of faster and more reproducible separations and measurements. As an early example of this trend [213], we show the separation of glycans extracted from a blood serum sample of a breast cancer patient (Figure 11).…”

“…It is currently hard to predict whether the desired improvements will come from the use of different derivatization schemes and separation conditions, any breakthrough developments in the CE-MS interfacing technologies, MS designs, or combined incremental improvements in all these areas. Various advances in CE-MS of glycoconjugates have been the subject of recent reviews [216, 225, 226]. …”