Accurate assessment of endogenous protein from human colon FFPE tissues by LC-MS/MS bioanalysis
Soumya Kandi, Ph.D.
Monday, June 16, 2025
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Chapters
Introduction
Additional authors and Acknowledgements
Mixed Linkage Kinase domain like (Mlkl) protein is a mediator of cell death in the necroptotic pathway
Protein bioanalysis from Formalin Fixed Paraffin Embedded (FFPE) Tissues
Analytical workflow for sample preparation and analysis
Instrumentation and LC/MS parameters for targeted protein analysis
FFPE tissue processing workflow
Endogenous Mlkl signature peptide detected from FFPE colon tissue
Determination of Mlkl protein expression from Formalin Fixed Paraffin Embedded (FFPE) colon resection and biopsy tissues
Proteomics analysis of digests from FFPE tissues
Several proteins are differentially expressed in UC resection vs. NH resection
Protein expression changes in UC grouped by biological processes
Endogenous Mlkl sig peptide is detectable with as low as 25-50mm2 FFPE tissue
Higher endogenous Mlkl signature peptide recovery from UC resection compared to Biopsy tissues; opposite trend observed with Biopsy samples
MLKL IHC: Stain Total Area And Intensity Are Greater in UC Colon
Endogenous Mlkl sig peptide is detectable with as low as 25-50mm2 FFPE tissue (< 10mm2 mucosal area)
Normalizing to mucosa serves as a biologically relevant normalization allowing for better data interpretation.
Summary
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