Mass Spectrometry Imaging of N‑Glycans from Formalin-Fixed Paraffin-Embedded Tissue Sections Using a Novel Subatmospheric Pressure Ionization Source
journal contributionposted on 2019-09-26, 11:34 authored by Yatao Shi, Zihui Li, Mildred A. Felder, Qinying Yu, Xudong Shi, Yajing Peng, Qinjingwen Cao, Bin Wang, Luigi Puglielli, Manish S. Patankar, Lingjun Li
N-linked glycosylation, featuring various glycoforms, is one of the most common and complex protein post-translational modifications (PTMs) controlling protein structures and biological functions. It has been revealed that abnormal changes of protein N-glycosylation patterns are associated with many diseases. Hence, unraveling the disease-related alteration of glycosylation, especially the glycoforms, is crucial and beneficial to improving our understanding about the pathogenic mechanisms of various diseases. In past decades, given the capability of in situ mapping of biomolecules and their region-specific localizations, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been widely applied to the discovery of potential biomarkers for many diseases. In this study, we coupled a novel subatmospheric pressure (SubAP)/MALDI source with a Q Exactive HF hybrid quadrupole-orbitrap mass spectrometer for in situ imaging of N-linked glycans from formalin-fixed paraffin-embedded (FFPE) tissue sections. The utility of this new platform for N-glycan imaging analysis was demonstrated with a variety of FFPE tissue sections. A total of 55 N-glycans were successfully characterized and visualized from a FFPE mouse brain section. Furthermore, 29 N-glycans with different spatial distribution patterns could be identified from a FFPE mouse ovarian cancer tissue section. High-mannose N-glycans exhibited elevated expression levels in the tumor region, indicating the potential association of this type of N-glycans with tumor progression.
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tumor progressionprotein N-glycosylation patternsFFPE mouse brain section29 N-glycansFormalin-Fixed Paraffin-Embedded Tissue Sectionsquadrupole-orbitrap mass spectrometerprotein structuresnovel subatmospheric pressureglycoformPTM55 N-glycanscancer tissue sectionQ Exactive HFprotein post-translational modificationsHigh-mannose N-glycansFFPE mouseN-linked glycansMALDI-MSIdisease-related alterationNovel Subatmospheric Pressure Ionization Source N-linked glycosylationexpression levelsdistribution patternsmass Spectrometry ImagingN-glycan imaging analysistumor regionFFPE tissue sectionstissue sectionsregion-specific localizationsformalin-fixed paraffin-embedded