Journal article

What are we missing by using hydrophilic enrichment? Improving bacterial glycoproteome coverage using total proteome and FAIMS analysis

Ameera Raudah Ahmad Izaham, Ching-Seng Ang, Shuai Nie, Lauren Bird, Nicholas Williamson, Nichollas Scott

Cold Spring Harbor Laboratory | Published : 2020

Abstract

ABSTRACT Hydrophilic Interaction Liquid Chromatography (HILIC) glycopeptide enrichment is an indispensable tool for the high-throughput characterisation of glycoproteomes. Despite its utility, HILIC enrichment is associated with a number of short comings including requiring large amounts of starting material, potentially introducing chemical artefacts such as formylation, and biasing/under-sampling specific classes of glycopeptides. Here we investigate HILIC enrichment independent approaches for the study of bacterial glycoproteomes. Using three Burkholderia species ( B. cenocepacia, B. dolosa and B. ubonensis ) we demonstrate that short aliphatic O -linked glycopeptides are typically absent..

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