Matrix assisted laser desorption ionization imaging mass spectrometry workflow for spatial profiling analysis of N-linked Glycan expression in tissues

Thomas W. Powers, E. Ellen Jones, Lucy R. Betesh, Patrick R. Romano, Peng Gao, John A. Copland, Anand S. Mehta, Richard R. Drake

Research output: Contribution to journalArticlepeer-review

80 Scopus citations

Abstract

A new matrix assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in tissues is described. Application of an endoglycosidase, peptide N-glycosidase F (PNGaseF), directly on tissues followed by incubation releases N-linked glycan species amenable to detection by MALDI-IMS. The method has been designed to simultaneously profile the multiple glycan species released from intracellular organelle and cell surface glycoproteins, while maintaining histopathology compatible preparation workflows. A recombinant PNGaseF enzyme was sprayed uniformly across mouse brain tissue slides, incubated for 2 h, then sprayed with 2,5-dihydroxybenzoic acid matrix for MALDI-IMS analysis. Using this basic approach, global snapshots of major cellular N-linked glycoforms were detected, including their tissue localization and distribution, structure, and relative abundance. Off-tissue extraction and modification of glycans from similarly processed tissues and further mass spectrometry or HPLC analysis was done to assign structural designations. MALDI-IMS has primarily been utilized to spatially profile proteins, lipids, drug, and small molecule metabolites in tissues, but it has not been previously applied to N-linked glycan analysis. The translatable MALDI-IMS glycan profiling workflow described herein can readily be applied to any tissue type of interest. From a clinical diagnostics perspective, the ability to differentially profile N-glycans and correlate their molecular expression to histopathological changes can offer new approaches to identifying novel disease related targets for biomarker and therapeutic applications.

Original languageEnglish (US)
Pages (from-to)9799-9806
Number of pages8
JournalAnalytical Chemistry
Volume85
Issue number20
DOIs
StatePublished - Oct 15 2013

ASJC Scopus subject areas

  • Analytical Chemistry

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