Structural Elucidation of N-Glycans in IgG and Human Serum Glycoprotein Through the Combination of Offline Two-dimensional Chromatography and Exoglycosidase Sequencing

李凤; 张含智; 康经武

doi:doi：10.3969/j.issn.1004-4957.2020.02.005

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Structural Elucidation of N-Glycans in IgG and Human Serum Glycoprotein Through the Combination of Offline Two-dimensional Chromatography and Exoglycosidase Sequencing

更新时间：2023-05-25

- Structural Elucidation of N-Glycans in IgG and Human Serum Glycoprotein Through the Combination of Offline Two-dimensional Chromatography and Exoglycosidase Sequencing
- Vol. 39, Issue 2, Pages: 198-204(2020)
- 作者机构：
  
  1. 西安文理学院化学工程学院
  2. 上海市食品药品检验所
  3. 中国科学院上海有机化学研究所生命有机化学国家重点实验室
- 作者简介：
- 基金信息：
- DOI：doi：10.3969/j.issn.1004-4957.2020.02.005
  CLC：
- Published：2020
- 稿件说明：
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李凤, 张含智, 康经武. Structural Elucidation of N-Glycans in IgG and Human Serum Glycoprotein Through the Combination of Offline Two-dimensional Chromatography and Exoglycosidase Sequencing[J]. 2020, 39(2): 198-204.
DOI：

李凤, 张含智, 康经武. Structural Elucidation of N-Glycans in IgG and Human Serum Glycoprotein Through the Combination of Offline Two-dimensional Chromatography and Exoglycosidase Sequencing[J]. 2020, 39(2): 198-204. DOI： doi：10.3969/j.issn.1004-4957.2020.02.005.

摘要

基于离线二维色谱和糖苷外切酶分析，建立了一种有效、可靠的糖蛋白N 糖链的结构鉴定方法。首先采用弱阴离子交换色谱实现不同唾液酸程度糖链的制备，随后将激光诱导毛细管电泳（CE-LIF）与糖苷外切酶相结合进行糖链结构鉴定。将分离制备后的糖链荧光标记后，进行自上而下的消化(top-down digestion)以及自下而上的结构归属(bottom-up identification)，共检测到IgG中的18种糖型。将上述策略进一步应用于人血清中N-糖链的分析，发现不同唾液酸化程度糖链的相对量为S1∶S2∶S3∶S4=4.7∶20.9∶6∶1。结果表明，人血清中高丰度的中性糖链与IgG的中性糖链结构非常相似，而血清中含量最丰富的是二唾液化两天线的N 糖型FA2G2S2。该方法在糖蛋白N-糖链的结构表征中具有一定的应用潜力。

Abstract

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.

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