Release Time:2020-06-13  Hits:

Indexed by: Journal Papers

Date of Publication: 2020-03-15

Journal: ANALYTICA CHIMICA ACTA

Included Journals: SCIE、EI、PubMed

Volume: 1102

Page Number: 130-139

ISSN: 0003-2670

Key Words: Immunoglobulin G; Fc-glycopeptide; HILIC; Purification; Absolute quantitation; Serum

Abstract: Absolute quantitation of IgG-1 Fc-glycosylation, which is crucial for the clinical practice of glycobiomarkers and quality control of biopharmaceuticals, has been hindered by the lack of glycopeptide standards. In this study, eleven high abundant IgG-1 Fc-glycopeptides with definite peptide sequences and glycoforms were purified from commercial IgG protein by using two-dimensional hydrophilic interaction liquid chromatographic system. Based on the acquired glycopeptide standards, an absolute quantitation strategy was developed to determine the concentrations of 11 target IgG-1 glycopeptides from pooled human sera. A wide range of Fc-glycopeptide concentrations from 0.60 to 17.61 nmol mL(-1) was achieved with excellent accuracy and reproducibility from pooled human sera IgG-1. Compared to conventional relative quantitation, this strategy provides more accurate distribution profiles of 11 high abundant Fc-glycopeptides and degree of glycosylation from pooled human sera IgG-1. (C) 2019 Elsevier B.V. All rights reserved.