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

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.