Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence

Szekrényes, Ákos and Park, SungAe Suhr and Santos, Marcia and Lew, Clarence and Jones, Aled and Haxo, Ted and Kimzey, Michael and Pourkaveh, Shiva and Szabó, Zoltán and Sosic, Zoran and Feng, Peng and Váradi, Csaba and de l'Escaille, François and Falmagne, Jean-Bernard and Sejwal, Preeti and Niedringhaus, Thomas and Michels, David and Freckleton, Gordon and Hamm, Melissa and Manuilov, Anastasiya and Schwartz, Melissa and Luo, Jiann-Kae and van Dyck, Jonathan and Leung, Pui-King and Olajos, Marcell and Gu, Yingmei and Gao, Kai and Wang, Wenbo and Wegstein, Jo and Tep, Samnang and Guttman, András (2016) Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence. MABS, 8 (1). pp. 56-64. ISSN 1942-0862

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Abstract

An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra highperformance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established.

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