ABSTRACT
Porous Graphitized Carbon nano-liquid chromatography tandem mass spectrometry (PGC-nLC-MS/MS) is a glycomics technique with the unique capacity to differentiate isobaric glycans. The lack of suitable software tools integrating chromatography and MS-information delivered by PGC-nLC-MS/MS has been limiting fast and robust glycan identification and quantitation. We report a LC-system-independent strategy called GlycoRRT that combines relative retention time (RRT) and negative ion fragment spectra analyses for isobaric structure-specific glycomics of PGC-nLC-MS/MS data. The GlycoRRT toolset is fully customizable and easily adaptable enabling semi-automated high-throughput structural assignments. The current library contains over 200 entries and their individual meta-data (MS instrumentation, experimental conditions, retention times, fragmentation profiles and glycan structural diagnostic ion features) relevant for reliable data analyses. The GlycoRRT workflow was employed to map the N- and O-glycome in blood group matched human plasma and urine as well as decipher Immunoglobulin (IgG) glycosylation features from 13 different animal species. We have also developed visualization tools to enable a consistent, reliable, and reproducible analysis of large sets of multidimensional PGC-nLC-MS/MS glycomics data. This comprehensive glycan resource provides the glycan map of human and animal species, will serve as a reference in dissecting the role of glycans in host pathogen interaction and zoonotic disease transmission.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
↵# Novo Nordisk, Novo Nordisk Park 2760 Måløv, Denmark
CREDIT AUTHOR STATEMENT
Kathirvel Alagesan – Conceptualization, Methodology, Software, Validation, Investigation, Formal analysis, data curation, Writing – Original draft, Writing – Review & Editing, Visualization, Project administration
Falko Schirmeister – Investigation, Software, Validation, Writing – Review & Editing, Visualization
Uwe Möginger – Investigation, Validation, Writing – Review & Editing, Visualization
Arun Everest-Dass - Validation, Writing – Review & Editing, Visualization
Friedrich Altmann – Resources, Writing – Review & Editing
Peter H Seeberger – Resources, Writing – Review & Editing
Mark von Itzstein – Resources, Writing – Review & Editing
Nicolle H Packer – Resources, Writing – Review & Editing
Daniel Kolarich – Conceptualization, Methodology, Resources, Visualization, Writing – Review & Editing, Funding acquisition