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A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations

Author:
Song, Ting, Aldredge, Danielle, Lebrilla, Carlito B.
Source:
Analytical chemistry 2015 v.87 no.15 pp. 7754-7762
ISSN:
1520-6882
Subject:
biomarkers, blood serum, genome, glycoproteins, glycosylation, humans, liquid chromatography, mass spectrometry, oligosaccharides, polysaccharides, post-translational modification, therapeutics, vaccines
Abstract:
Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. Even now, the exact structures of many abundant N-glycans in serum are not known. Structural elucidation of oligosaccharides remains difficult and time-consuming. Here, we introduce a means of rapidly identifying released N-glycan structures using their accurate masses and retention times based on a glycan library. This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. The method employs primarily nanoflow liquid chromatography and accurate mass spectrometry. The method allows us to readily profile N-glycans in biological fluids with deep structural analysis. This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood.
Agid:
5300027