Work

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

Year: 2026

Type: article

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

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104336

Authors: Kevan T. Knizner, Darren T. Gass, Yasuyuki Matsumoto, Uriel Ortega-Rodriguez, John Q. Bettinger, Tongzhong Ju

Institutions: United States Food and Drug Administration, Center for Drug Evaluation and Research

Language: English

Topic: Glycosylation and Glycoproteins Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550