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Accueil > Archives > Articles Scientifiques > Spectrométrie de Masse

Mass spectrometric quantification of N-linked glycans by reference to exogenous standards

par Emmanuel Maes - publié le , mis à jour le

Voici un article qui nous est recommandé à la lecture par Mr Y Guérardel. Il est paru dans Journal of proteome Research par Mehta et al du CCRC
Il s’agit de quantifier les N-Glycans grâce à des standards polyhexoses permethylés avec de l’iodure de methyl marqué 13C.

Mehta N. et al, 2016, J. Proteome Res., 15, 2969-80
Mehta N. et al, 2016, J. Proteome Res., 15, 2969-80

Voici l’abstract :
" Environmental and metabolic processes shape the profile of glycoprotein glycans expressed by cells, whether in culture, developing tissues, or mature organisms. Quantitative characterization of glycomic changes associated with these conditions has been achieved historically by reductive coupling of oligosaccharides to various fluorophores following release from glycoprotein and subsequent HPLC or capillary electrophoretic separation. Such labeling-based approaches provide a robust means of quantifying glycan amount based on fluorescence yield. Mass spectrometry, on the other hand, has generally been limited to relative quantification in which the contribution of the signal intensity for an individual glycan is expressed as a percent of the signal intensity summed over the total profile. Relative quantification has been valuable for highlighting changes in glycan expression between samples ; sensitivity is high, and structural information can be derived by fragmentation. We have investigated whether MS-based glycomics is amenable to absolute quantification by referencing signal intensities to well-characterized oligosaccharide standards. We report the qualification of a set of N-linked oligosaccharide standards by NMR, HPLC, and MS. We also demonstrate the dynamic range, sensitivity, and recovery from complex biological matrices for these standards in their permethylated form. Our results indicate that absolute quantification for MS-based glycomic analysis is reproducible and robust utilizing currently available glycan standards.
KEYWORDS : quantification, N-glycan, permethylation, standard, mass spectrometry""

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