GlycoFASP: A Universal Method to Prepare Complex Mixtures for O-Glycoproteomic Analysis

03 April 2025, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Here we describe a method titled filter-aided preparation of O-glycoproteomic samples (GlycoFASP), which takes advantage of molecular-weight cut-off filters and O-glycoprotease digestion specificity. Using GlycoFASP, preparation for MS analysis of biological samples is completed within one day and only requires one milligram of sample. It is both low-cost and amenable to non-specialists. GlycoFASP regularly achieves depletion of nearly all non-O-glycosylated proteins, with ≥70% of the protein signal originating from O-glycoproteins.

Keywords

mass spectrometry
glycobiology
glycoproteomics
FASP

Supplementary materials

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Supplemental Data
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SI Figures 1-3, SI Tables 1-3
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