DIA-MS proteome analysis of formalin-fixed paraffin-embedded glioblastoma tissues

Kenneth Weke, Sachin Kote*, Jakub Faktor, Sofian Al Shboul, Naomi Uwugiaren, Paul M. Brennan, David R. Goodlett, Ted R. Hupp, Irena Dapic*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Developments in quantitative proteomics and data-independent acquisition (DIA) methodology is enabling quantification of proteins in biological samples. Currently, there are a few reports on DIA mass spectrometry (MS) approaches for proteome analysis of formalin-fixed paraffin-embedded (FFPE) tissues. Therefore, to facilitate detection and quantification of immune- and glioblastoma (GBM)-relevant proteins from FFPE patient materials, we established a simple and precise DIA-MS workflow. We first evaluated different lysis buffers for their efficiency in protein extractions from FFPE GBM tissues. Our results showed that more than 1700 proteins were detected and over 1400 proteins were quantified from GBM FFPE tissue microdissections. GBM-relevant proteins (e.g., GFAP, FN1, VIM, and MBP) were quantified with high precision (median coefficient of variation <12%). In addition, immune-related proteins (e.g., ILF2, MIF, and CD38) were consistently detected and quantified. The strategy holds great potential for routinizing protein quantification in FFPE tissue samples.

Original languageEnglish
Article number339695
JournalAnalytica Chimica Acta
Early online date12 Mar 2022
Publication statusPublished - 29 Apr 2022

Keywords / Materials (for Non-textual outputs)

  • Data-independent acquisition
  • Quantitative proteomics
  • Formalin-fixed paraffin-embedded
  • Glioblastoma
  • Mass spectrometry


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