N-Glycan matrix-assisted laser desorption/ionization mass spectrometry imaging protocol for formalin-fixed paraffin-embedded tissues

Matthew T Briggs; Yin Ying Ho; Gurjeet Kaur; Martin K Oehler; Arun V Everest-Dass; Nicolle H Packer; Peter Hoffmann

doi:10.1002/rcm.7845

N-Glycan matrix-assisted laser desorption/ionization mass spectrometry imaging protocol for formalin-fixed paraffin-embedded tissues

Rapid Commun Mass Spectrom. 2017 May 30;31(10):825-841. doi: 10.1002/rcm.7845.

Authors

Matthew T Briggs^{1

2}, Yin Ying Ho¹, Gurjeet Kaur³, Martin K Oehler^{4

5}, Arun V Everest-Dass^{6

7}, Nicolle H Packer^{6

7}, Peter Hoffmann^{1

2}

Affiliations

¹ Adelaide Proteomics Centre, School of Biological Sciences, University of Adelaide, Adelaide, Australia, 5005.
² Institute for Photonics and Advanced Sensing (IPAS), University of Adelaide, Adelaide, Australia, 5005.
³ Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Pulau Pinang, Malaysia.
⁴ Department of Gynaecological Oncology, Royal Adelaide Hospital, Adelaide, South Australia, 5005, Australia.
⁵ Robinson Institute, University of Adelaide, Adelaide, Australia, 5005.
⁶ ARC Centre for Nanoscale BioPhotonics (CNBP), University of Adelaide, Adelaide, Australia, 5005.
⁷ Biomolecular Frontiers Research Centre, Macquarie University, Sydney, Australia, 2109.

PMID: 28271569
DOI: 10.1002/rcm.7845

Abstract

Rationale: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) of the proteome of a tissue has been an established technique for the past decade. In the last few years, MALDI-MSI of the N-glycome has emerged as a novel MALDI-MSI technique. To assess the accuracy and clinical significance of the N-linked glycan spatial distribution, we have developed a method that utilises MALDI-MSI followed by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS) in order to assign glycan structures to the differentiating MALDI-MSI glycan masses released from the tissue glycoproteins.

Methods and results: Our workflow presents a comprehensive list of instructions on how to (i) apply MALDI-MSI to spatially map the N-glycome across formalin-fixed paraffin-embedded (FFPE) clinical samples, (ii) structurally characterise N-glycans extracted from consecutive FFPE tissue sections by LC/MS/MS, and (iii) match relevant N-glycan masses from MALDI-MSI with confirmed N-glycan structures determined by LC/MS/MS.

Conclusions: Our protocol provides groups that are new to this technique with instructions how to establish N-glycan MALDI-MSI in their laboratory. Furthermore, the method assigns N-glycan structural detail to the masses obtained in the MALDI-MS image. Copyright © 2017 John Wiley & Sons, Ltd.