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Comparative Analysis of Different N -glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry by Yudong Guan; Min Zhang; Jigang Wang; Hartmut Schlüter is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Comparative Analysis of Different N -glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry about?

Protein N-glycosylation characterization is challenging due to structural micro-and macro-heterogeneity. Although various N-glycan preparation strategies, including purification and derivatization, have been previously developed prior to mass spectrometric analysis, systematic evaluation still needs to be performed. This study compared the different N-glycan purification strategies, including filter-aided sample preparation, de-N-glycosylated protein precipitation, and trypsin digestion followed by reversed phase-based solid-phase extraction, and derivatization approaches, such as solid-phase permethylation, reductive amination, and reduction. With the comparative analysis, an optimized solid-phase permethylation (OSPP) workflow was developed for mass spectrometric N-glycomics, showing simplified analysis for Nglycan compositions and high yields using etanercept. The N-glycan samples released from trastuzumab and adalimumab were utilized to test OSPP to obtain their N-glycan profiles using mass spectrometry. Based on different standard procedures across laboratories, this study provides the reference for analysts to select an appropriate N-glycan preparation method with their resea

Who reads Comparative Analysis of Different N -glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry?

Author

Yudong Guan; Min Zhang; Jigang Wang; Hartmut Schlüter

Publisher

American Chemical Society (ACS)

Published

2021

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)