Quantitative Glycomics Strategies

Abstract

The correlations between protein glycosylation and many biological processes and diseases are increasing the demand for quantitative glycomics strategies enabling sensitive monitoring of changes in the abundance and structure of glycans. This is currently attained through multiple strategies employing several analytical techniques such as capillary electrophoresis, liquid chromatography, and mass spectrometry. The detection and quantification of glycans often involve labeling with ionic and/or hydrophobic reagents. This step is needed in order to enhance detection in spectroscopic and mass spectrometric measurements. Recently, labeling with stable isotopic reagents has also been presented as a very viable strategy enabling relative quantitation. The different strategies available for reliable and sensitive quantitative glycomics are herein described and discussed. Molecular & Cellular Proteomics

Evaporative fluorophore labeling of carbohydrates via reductive amination

“…Glycosylation is one of the major and most complicated forms of post-translational modifications that plays essential roles in the activity and effector function of monoclonal antibodies [3] demanding improved glycoanalytical methods. Frequently 3/13 used techniques for carbohydrate analysis are mass spectrometry (MS) [4], liquid chromatography (LC) [5,6], capillary electrophoresis (CE) [7][8][9], some combination of them [10][11][12] and if necessary, nuclear magnetic resonance spectroscopy (NMR) [13,14].…”

Mass spectrometry-based N-linked glycomic profiling as a means for tracking pancreatic cancer metastasis

“…3 Mass spectrometry (MS), on the other hand, offers an alternative with its high sensitivity and peak resolution for carbohydrate identification; 4 MALDI-TOF MS, in particular, is being developed as a platform for the qualitative and quantitative analysis of glycoconjugates. 5 Herein, we propose to utilize a MS-based analytical method to investigate the surface-bound-N-glycan profiles for three pancreatic cancer cell linesdrepresentative of different pancreatic cancer locationsdand a normal pancreatic cell line. The membrane fractionations of pancreatic cancer and normal pancreatic cells are enzymatically deglycosylated and subsequently permethylated via a quantitative solid-phase method to obtain corresponding N-glycans.…”

Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry: Mechanistic Studies and Methods for Improving the Structural Identification of Carbohydrates

“…Various methods for derivatization have been developed, such as uses of hydrazines 129) and other labeling techniques. 130) Although derivatization provides advantages, conventional derivatization typically requires harsh conditions (strong base) and long reaction times. Most derivatization methods require clean-up procedures for the removal of chemicals, resulting in a loss of samples, which especially leads to a serious problem for trace analysis.…”