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Mol Cell Proteomics. 2017 Apr;16(4):524-536. doi: 10.1074/mcp.M116.062414. Epub 2017 Jan 25.

Unlocking Cancer Glycomes from Histopathological Formalin-fixed and Paraffin-embedded (FFPE) Tissue Microdissections.

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From the ‡Max Planck Institute of Colloids and Interfaces, Department of Biomolecular Systems, 14424 Potsdam, Germany.
§Freie Universität Berlin, Department of Biology, Chemistry, Pharmacy, Institute of Chemistry and Biochemistry, 14195 Berlin, Germany.
¶Faculty of Science, Department of Biology, Division of Molecular Biology, University of Zagreb, Zagreb, Croatia.
‖Institute for Pathology and Cytology, University Hospital Merkur, Zagreb, Croatia.
**Department of Pathology, Medical School Zagreb, University of Zagreb, Zagreb, Croatia.
From the ‡Max Planck Institute of Colloids and Interfaces, Department of Biomolecular Systems, 14424 Potsdam, Germany;


N- and O-glycans are attractive clinical biomarkers as glycosylation changes in response to diseases. The limited availability of defined clinical specimens impedes glyco-biomarker identification and validation in large patient cohorts. Formalin-fixed paraffin-embedded (FFPE) clinical specimens are the common form of sample preservation in clinical pathology, but qualitative and quantitative N- and O-glycomics of such samples has not been feasible to date. Here, we report a highly sensitive and glycan isomer selective method for simultaneous N- and O-glycomics from histopathological slides. As few as 2000 cells isolated from FFPE tissue sections by laser capture microdissection were sufficient for in-depth histopathology-glycomics using porous graphitized carbon nanoLC ESI-MS/MS. N- and O-glycan profiles were similar between unstained and hematoxylin and eosin stained FFPE samples but differed slightly compared with fresh tissue. This method provides the key to unlock glyco-biomarker information from FFPE histopathological tissues archived in pathology laboratories worldwide.

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