1. Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence
- Author
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Jo Wegstein, Preeti Sejwal, François de l’Escaille, Jonathan van Dyck, Ákos Szekrényes, András Guttman, Anastasiya Manuilov, Kai Gao, Gordon Freckleton, Wenbo Wang, David A. Michels, Marcia Santos, Clarence Lew, Zoltan Szabo, Ted Haxo, Michael Kimzey, Peng Feng, Jean Bernard Falmagne, Samnang Tep, Pui King Leung, Aled Jones, Jiann Kae Luo, Yingmei Gu, Csaba Váradi, Melissa Hamm, Thomas Niedringhaus, Melissa Schwartz, Zoran Sosic, Sung Ae Suhr Park, Marcell Olajos, and Shiva Pourkaveh
- Subjects
Glycan ,Immunology ,Analytical chemistry ,N-glycans ,02 engineering and technology ,01 natural sciences ,Fluorescence ,Capillary electrophoresis ,Biotherapeutics ,Polysaccharides ,Report ,Humans ,Immunology and Allergy ,Sample preparation ,Laser-induced fluorescence ,Chromatography, High Pressure Liquid ,Mapping study ,Peak area ,Reproducibility ,Chromatography ,biology ,Chemistry ,Lasers ,010401 analytical chemistry ,Multi site ,Electrophoresis, Capillary ,Intercompany study ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,biology.protein ,0210 nano-technology - 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 high-performance 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.
- Published
- 2015
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