Protocol for simultaneous profiling of N- and O-glycans on glycoproteins using a one-pot format

STAR Protocols 2026 Open access

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Authors: Kevan T. Knizner, Darren T. Gass, Yasuyuki Matsumoto, Uriel Ortega-Rodriguez, John Q. Bettinger, Tongzhong Ju
DOI: 10.1016/j.xpro.2025.104336
Cited by: 1
References: 25

Abstract

Here, we present a protocol to simultaneously release and profile N - and O -glycans from glycoproteins in a one-pot format. We describe steps for protein reduction, alkylation, N -glycan release, Proteinase K digestion, HILIC enrichment, N -glycan reduction, N- / O -glycan permethylation while releasing O -glycans, and analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). For complete details on the use and execution of this protocol, please refer to Ortega-Rodriguez et al. 1

Fields of Study & Subjects

Glycosylation and Glycoproteins Research Advanced Proteomics Techniques and Applications Carbohydrate Chemistry and Synthesis

Publication details

Journal / source
STAR Protocols
Publisher
Elsevier BV
Volume
7
Issue
1
Pages
104336–104336
Publication date
5 February 2026
Language
EN