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
| Autor: | Miao He, Amanda M. Ackermann, Eva Morava, Jie Chen, Andrew C. Edmondson, Michael J. Bennett, Kosuke Izumi, Xueli Li, Gail Ditewig Meyers, Can Ficicioglu |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
PNGase F chemistry.chemical_classification Glycan Glycosylation biology Electrospray ionization Biochemistry (medical) Clinical Biochemistry 030105 genetics & heredity Mass spectrometry Glycopeptide 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry Biochemistry Transferrin biology.protein Glycoprotein |
| Zdroj: | Clinical Chemistry. 65:653-663 |
| ISSN: | 1530-8561 0009-9147 |
| Popis: | 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 CONCLUSIONS The clinical specificity and sensitivity of N-glycan analysis was much improved with this method. Additional CDGs can be diagnosed that would be missed by carbohydrate-deficient transferrin analysis. The assay provides novel biomarkers with diagnostic and potentially therapeutic significance. |
| Databáze: | OpenAIRE |
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