ArticleAnalytical chemistry2016
Structural Characterization of Serum N-Glycans by Methylamidation, Fluorescent Labeling, and Analysis by Microchip Electrophoresis.
Article in Analytical chemistry, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 49 citations in OpenAlex.
- Autoimmune hepatitis displays distinctively high multi-antennary sialylation on plasma N-glycans compared to other liver diseases.Journal of translational medicine · 2024Article
- Structure and function of microbial α-l-fucosidases: a mini review.Essays in biochemistry · 2023Review
- The cancer glycocode as a family of diagnostic biomarkers, exemplified by tumor-associated gangliosides.Frontiers in oncology · 2023Review
- Quantitative clinical glycomics strategies: A guide for selecting the best analysis approach.Mass spectrometry reviews · 2022Review
- Fractionation and characterization of sialyl linkage isomers of serum N-glycans by CE-MS.Journal of separation science · 2022Article
- High-throughput N-glycan screening method for therapeutic antibodies using a microchip-based DNA analyzer: a promising methodology for monitoring monoclonal antibody N-glycosylation.Analytical and bioanalytical chemistry · 2021Article
- Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis.Experientia supplementum (2012) · 2021Article
- Exosome-Mediated Crosstalk between Keratinocytes and Macrophages in Cutaneous Wound Healing.ACS nano · 2020Article
- In-Depth Compositional and Structural Characterization of N-Glycans Derived from Human Urinary Exosomes.Analytical chemistry · 2019Article
- Expanding the Reaction Space of Linkage-Specific Sialic Acid Derivatization.Molecules (Basel, Switzerland) · 2019Article
- Vitronectins produced by human cirrhotic liver and CClFEBS open bio · 2019Article
- Profiling the N-Glycan Composition of IgG with Lectins and Capillary Nanogel Electrophoresis.Analytical chemistry · 2019Article
- Measurement of Neutral and Sialylated IgG N-Glycome at Asn-297 by CE-LIF to Assess Hypogalactosylation in Rheumatoid Arthritis.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Capillary Electrophoresis Separations of Glycans.Chemical reviews · 2018Review
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- Article
- Capillary electrophoresis with stationary nanogel zones of galactosidase and Erythrina cristagalli lectin for the determination of β(1-3)-linked galactose in glycans.Journal of chromatography. A · 2017Article
- Capillary electrophoresis-mass spectrometry for direct structural identification of serum N-glycans.Journal of chromatography. A · 2017Article
- Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes.Analytical chemistry · 2017Article
- Recent Advances in the Analysis of Complex Glycoproteins.Analytical chemistry · 2017Review
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
To characterize the structures of N-glycans derived from human serum, we report a strategy that combines microchip electrophoresis, standard addition, enzymatic digestion, and matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). We compared (i) electrophoretic mobilities of known N-glycans from well-characterized (standard) glycoproteins through standard addition, (ii) the electrophoretic mobilities of N-glycans with their molecular weights determined by MALDI-MS, and (iii) electrophoretic profiles of N-glycans enzymatically treated with fucosidase. The key step to identify the sialylated N-glycans was to quantitatively neutralize the negative charge on both α2,3- and α2,6-linked sialic acids by covalent derivatization with methylamine. Both neutralized and nonsialylated N-glycans from these samples were then reacted with 8-aminopyrene-1,3,6-trisulfonic acid (APTS) to provide a fluorescent label and a triple-negative charge, separated by microchip electrophoresis, and detected by laser-induced fluorescence. The methylamidation step leads to a 24% increase in the peak capacity of the separation and direct correlation of electrophoretic and MALDI-MS results. In total, 37 unique N-glycan structures were assigned to 52 different peaks recorded in the electropherograms of the serum samples. This strategy ensures the needed separation efficiency and detectability, easily resolves linkage and positional glycan isomers, and is highly reproducible.
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