Glycan analysis by isobaric aldehyde reactive tags and mass spectrometry.

ABSTRACT

Glycans play significant roles in physiological and pathological processes. Therefore, quantitative analysis of glycans from normal and disease specimens can provide insight into disease onset and progression. Relative glycan quantification usually requires modification of the glycans with either chromogenic or fluorogenic tags for optical measurement or isotopic tags for mass spectrometric analysis. Because of rapid advances in mass spectrometry (MS) instruments in resolution, sensitivity, and speed, MS-based methods have become increasingly popular for glycan analysis in the past decade. However, current isotopic tags for glycan labeling are mostly mass-shift tags generating mass differences in precursor ions for quantification, which can complicate mass spectra. In this study, we report the synthesis and characterization of isobaric aldehyde reactive tags (iARTs) for glycan quantification using tandem MS. We applied iARTs to the relative identification and quantification of glycans of gp120, a glycoprotein from human immunodeficiency virus. The results show that iARTs provide strong signals for glycan identification. Although we only show the synthesis and characterization of two iARTs reagents, iARTs can be readily expanded to six-plex tags for quantitative analysis of six samples concurrently.