Simultaneous Analysis of Sulfated and Phosphorylated Glycans by Serotonin-Immobilized Column Enrichment and Hydrophilic Interaction Chromatography

Changes in the structures and quantities of sulfated glycans play important roles in inflammatory and neurological diseases and cancer. Therefore, sulfated glycans are expected to become diagnostic markers for a variety of diseases such as Alzheimer’s disease and cancer. On the other hand, structura...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2018-07, Vol.90 (14), p.8387-8395
Main Authors: Yamada, Keita, Kayahara, Haruna, Kinoshita, Mitsuhiro, Suzuki, Shigeo
Format: Article
Language:English
Subjects:
Abnormalities
Cancer
Chemistry
Chromatography
Desalination
Diagnostic systems
Gastric cancer
Lysosomal enzymes
N-glycans
Neurological diseases
Neurological disorders
Ovalbumin
Phosphorylation
Polysaccharides
Proteins
Quantitative analysis
Serotonin
Sulfates
Thyroglobulin
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Summary:Changes in the structures and quantities of sulfated glycans play important roles in inflammatory and neurological diseases and cancer. Therefore, sulfated glycans are expected to become diagnostic markers for a variety of diseases such as Alzheimer’s disease and cancer. On the other hand, structural abnormalities in the phosphorylated glycans on lysosomal enzymes cause a number of lysosomal diseases, while novel phosphorylated glycans have been found in other proteins. As with sulfated glycans, structural and quantitative changes in these phosphorylated glycans and their associations with disease are also of interest. In this article, we introduce a new method for the simultaneous analysis of sulfated and phosphorylated glycans. We first employ a serotonin-immobilized column to enrich these glycans. Glycans obtained in this manner were sequentially subjected to other analytical techniques without desalting. We employed hydrophilic interaction chromatography to distinguish the sulfate and phosphate groups of the glycans and were able to analyze sulfated and phosphorylated N-glycans expressed on thyroglobulin, ovalbumin, and myozyme. We showed that our method not only analyzes sulfated and phosphorylated glycans, but also glycans containing the GlcNAc-HPO3 residue. We comparatively analyzed sulfated glycans, phosphorylated glycans, and GlcNAc-HPO3-residue-containing glycans expressed on MKN7 cells (well-differentiated gastric cancer cells) and MKN45 cells (poorly differentiated gastric cancer cells). To the best of our knowledge, this is the first report of a method for the simultaneous and quantitative analysis of sulfated and phosphorylated glycans.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.8b00714