Combining glucose units, m/z, and collision cross section values

multiattribute data for increased accuracy in automated glycosphingolipid glycan identifications and its application in triple negative breast cancer

Katherine Wongtrakul-Kish*, Ian Walsh, Lyn Chiin Sim, Amelia Mak, Brian Liau, Vanessa Ding, Noor Hayati, Han Wang, Andre Choo, Pauline M. Rudd, Terry Nguyen-Khuong

*Corresponding author for this work

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Glycan head-groups attached to glycosphingolipids (GSLs) found in the cell membrane bilayer can alter in response to external stimuli and disease, making them potential markers and/or targets for cellular disease states. To identify such markers, comprehensive analyses of glycan structures must be undertaken. Conventional analyses of fluorescently labeled glycans using hydrophilic interaction high-performance liquid chromatography (HILIC) coupled with mass spectrometry (MS) provides relative quantitation and has the ability to perform automated glycan assignments using glucose unit (GU) and mass matching. The use of ion mobility (IM) as an additional level of separation can aid the characterization of closely related or isomeric structures through the generation of glycan collision cross section (CCS) identifiers. Here, we present a workflow for the analysis of procainamide-labeled GSL glycans using HILIC-IM-MS and a new, automated glycan identification strategy whereby multiple glycan attributes are combined to increase accuracy in automated structural assignments. For glycan matching and identification, an experimental reference database of GSL glycans containing GU, mass, and CCS values for each glycan was created. To assess the accuracy of glycan assignments, a distance-based confidence metric was used. The assignment accuracy was significantly better compared to conventional HILIC-MS approaches (using mass and GU only). This workflow was applied to the study of two Triple Negative Breast Cancer (TNBC) cell lines and revealed potential GSL glycosylation signatures characteristic of different TNBC subtypes.

Original languageEnglish
Pages (from-to)9078-9085
Number of pages8
JournalAnalytical Chemistry
Volume91
Issue number14
DOIs
Publication statusPublished - 16 Jul 2019
Externally publishedYes

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