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Glycoqueuing: Isomer-Specific Quantification for Sialylation-Focused Glycomics.
Analytical Chemistry
|July 23, 2019
Summary
A new glycoqueuing method enables precise quantification of sialylglycan isomers using mass spectrometry. This technique advances disease biomarker discovery by accurately analyzing complex glycan structures.
Area of Science:
- Glycomics
- Analytical Chemistry
- Biomarker Discovery
Background:
- Sialylglycans, specifically α-2,3- and α-2,6-linked sialic acids (SAs), are implicated in various diseases.
- Accurate isomeric quantification of sialylglycans is challenging due to their complexity, instability, and low MS detection sensitivity.
Purpose of the Study:
- To develop a novel analytical strategy for high-sensitivity, isomer-specific quantification of sialylglycans.
- To validate the method using fetal bovine serum (FBS) N-linked sialylglycans and apply it to disease biomarker research.
Main Methods:
- Introduced 'glycoqueuing,' a sequential chromatographic elution strategy.
- Utilized isotopic labeling followed by online reversed-phase high-performance liquid chromatography coupling with mass spectrometry (RP-HPLC-MS).
- Validated with detailed structural identification and quantification of FBS N-linked sialylglycan isomers, including branching isomers and Neu5Gc-containing glycans.
Main Results:
- Successfully differentiated numerous branching sialylglycan isomers.
- Analyzed 28 sialylglycan compositions containing Neu5Gc residues.
- Revealed a 55% increase in α-2,6-sialylated fucosylated N-glycans in hepatocellular carcinoma serum samples.
Conclusions:
- The glycoqueuing method provides high-sensitivity, isomer-specific quantification of sialylglycans.
- Demonstrated applicability in comparing N-glycans between different species (bovine vs. rabbit IgG).
- Highlights potential clinical utility for identifying serum sialylglycan biomarker candidates for diseases like hepatocellular carcinoma.
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