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Author Giorgetti, Jeremie ♦ D’Atri, Valentina ♦ Canonge, Julie ♦ Lechner, Antony ♦ Guillarme, Davy ♦ Colas, Olivier ♦ Wagner-Rousset, Elsa ♦ Beck, Alain ♦ Leize-Wagner, Emmanuelle ♦ François, Yannis-Nicolas
Source Hyper Articles en Ligne (HAL)
Content type Text
File Format PDF
Language English
Subject Keyword chim ♦ Chemical Sciences/Analytical chemistry
Abstract Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). As a consequence, glycosylation profiling must be deeply characterized. For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. However, no CE-ESI-MS approach has been assessed and validated. Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of N-glycosylation of mAbs at the glycopeptide level. Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labeled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision.
ISSN 00399140
Educational Use Research
Learning Resource Type Article
Publisher Date 2018-02-01
Journal Talanta
Volume Number 178
Page Count 8
Starting Page 530
Ending Page 537