Increased Clinical Sensitivity and Specificity of Plasma Protein N-Glycan Profiling for Diagnosing Congenital Disorders of Glycosylation by Use of Flow Injection-Electrospray Ionization-Quadrupole Time-of-Flight Mass Spectrometry
Increased Clinical Sensitivity and Specificity of Plasma Protein N-Glycan Profiling for Diagnosing Congenital Disorders of Glycosylation by Use of Flow Injection-Electrospray Ionization-Quadrupole Time-of-Flight Mass Spectrometry
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DOI：
10.1373/clinchem.2018.296780
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发表时间：
2019-05-01
影响因子：
9.3
通讯作者：
He, Miao
中科院分区：
文献类型：
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作者：
Chen, Jie;Li, Xueli;He, Miao
背景技术背景：先天性糖基化障碍（CDG）是迄今为止发现的最大的代谢障碍类型之一，有超过130种亚型。大多数CDG亚型是N-连接糖基化的病症，其中碳水化合物残基，即N-聚糖，在肽中与天冬酰胺分子后连接。为了提高CDG的诊断能力，我们开发并验证了使用流动注射-电喷雾电离-四极杆飞行时间质谱法的血浆N-聚糖测定。在PNGase F消化血浆糖蛋白后，使用还原末端的瞬时胺基将N-聚糖连接至喹诺酮，通过亲水相互作用色谱柱分离，结果：该方法与其他N-聚糖谱分析方法不同，因为它不受循环中游离聚糖的污染，是半定量的。检测的二唾液酸双触角N-聚糖检测范围下限为63 nmol/L。正常血浆中的大多数N-聚糖
BACKGROUND: Congenital disorders of glycosylation (CDG) represent 1 of the largest groups of metabolic disorders with >130 subtypes identified to date. The majority of CDG subtypes are disorders of N-linked glycosylation, in which carbohydrate residues, namely, N-glycans, are posttranslationally linked to asparagine molecules in peptides. To improve the diagnostic capability for CDG, we developed and validated a plasma N-glycan assay using flow injection-electrospray ionization-quadrupole time-of-flight mass spectrometry.METHODS: After PNGase F digestion of plasma glycoproteins, N-glycans were linked to a quinolone using a transient amine group at the reducing end, isolated by a hydrophilic interaction chromatography column, and then identified by accurate mass and quantified using a stable isotope-labeled glycopeptide as the internal standard.RESULTS: This assay differed from other N-glycan profiling methods because it was free of any contamination from circulating free glycans and was semiquantitative. The low end of the detection range tested was at 63 nmol/L for disialo-biantennary N-glycan. The majority of N-glycans in normal plasma had