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

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

Abstract read

In one paragraph

Article in STAR protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

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4 · The record

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5 · Who and what money

Authors and funding

6 authors.

Kevan T KniznerUnited States Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Silver Spring MD 20993, USA.

Darren T GassUnited States Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Silver Spring MD 20993, USA.

Yasuyuki MatsumotoUnited States Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Silver Spring MD 20993, USA.

Uriel Ortega-RodriguezUnited States Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Silver Spring MD 20993, USA.

John Q BettingerUnited States Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Silver Spring MD 20993, USA.

Tongzhong JuUnited States Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Silver Spring MD 20993, USA. Electronic address: tongzhong.ju@fda.hhs.gov.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

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.

Indexed as

GlycoproteinsPolysaccharidesSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationGlycosylationGlycoproteinsPolysaccharidesChemistryClassification DescriptionMass CytometryProtein Biochemistry

Identifiers

PMID41649929

DOI10.1016/j.xpro.2025.104336

PMCPMC12907076

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.