Structural Elucidation of N-Glycans in IgG and Human Serum Glycoprotein Through the Combination of Offline Two-dimensional Chromatography and Exoglycosidase Sequencing
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KeyWord:offline two-dimensional chromatography  capillary electrophoresis  immunoglobulin G(IgG)  N-glycan  human serum
LI Feng,ZHANG Han-zhi,KANG Jing-wu 1.School of Chemical Engineering,Xi'an University;2.Shanghai Institute for Food and Drug Control;3.State Key Laboratory of Bioorganic and Natural Products Chemistry,Shanghai Institute of Organic Chemistry,Chinese Academy of Sciences
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      A reliable and efficient method for the determination of N-glycan structures in glycoprotein was developed by the combination of two dimensional offline liquid chromatography and exoglycosidase sequencing.Firstly, the N-glycans at different sialylation degrees were prepared by weak anion exchange chromatography,then the structures of analytes were identified by capillary electrophoresis-laser induced fluorescence detection(CE-LIF) combined with exoglycosidase digestion.After fluorescent labellling,the structural annotation was performed by the top-down digestion and the bottom-up identification,and a total of 18 N-glycan structures in Immunoglobulin G(IgG) were identified.Finally,the same method was applid in the analysis of N-glycans in human serum samples.The ratio for monosialylated,disialylated,trisialylated and tetrasialylated degree was 4.7∶20.9∶6∶1Results showed that high abundance neutral sugar chains in human serum were very similar to those in IgG in structure,and the disialylated N-glycan structure FA2G2S2 was the most abaundent in serum N-glycan pool.The method had an application potential in N-glycosylation analysis.