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CROSS-LABORATORY VALIDATION OF GLYCOSPHINGOLIPIDS PROFILING IN PDAC USING HIGH-RESOLUTION ION MOBILITY MASS SPECTROMETRY

Authors: Šubrtová Veronika | Giang Vo Huong | Jirásko Robert | Peterka Ondřej | Vošmik Milan | Bindila Laura | Holčapek Michal

Year: 2025

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	CROSS-LABORATORY VALIDATION OF GLYCOSPHINGOLIPIDS PROFILING IN PDAC USING HIGH-RESOLUTION ION MOBILITY MASS SPECTROMETRY	Glycosphingolipids (GSLs) are membrane-bound molecules involved in cell signaling, immune responses, and host–pathogen interactions. Their structural alterations are linked to various diseases, such as lysosomal disorders, neurodegeneration, and cancer. Accurate structural characterization and quantification of GSLs are essential for understanding their biological functions, but their amphiphilic nature and low abundance in complex biological matrices pose comprehensive analysis technically demanding. Mass spectrometry has significantly advanced GSL analysis by improving sensitivity and structural elucidation, but untargeted lipidomics often fails to detect low-abundance species due to insufficient enrichment and complex sample preparation. Recent developments, including ion mobility spectrometry (IMS-MS) and multidimensional techniques like LC-TIMS-PASEF, have shown promise in resolving structural isomers and improving throughput. However, GSL profiling still requires optimized extraction protocols and high-resolution methods to achieve comprehensive coverage, particularly in clinical samples. This approach, developed by Huong Giang Vo et al.1, has been applied in clinical studies of Parkinson’s disease. The aim of this work was to apply this workflow to analyze glycosphingolipids in serum samples from patients with pancreatic ductal adenocarcinoma (PDAC) and to perform a cross-laboratory comparison between two research groups. Initial sample preparation and measurements were conducted in the laboratory of Dr. Laura Bindila (Germany) using reversed-phase liquid chromatography (RP-LC) coupled with high-resolution trapped ion mobility mass spectrometry (timsTOF Pro, Bruker) in negative ion mode. Identical extracts were subsequently analyzed in the laboratory of Prof. Michal Holčapek (Czech Republic) using a newer TIMS instrument (timsTOF Ultra, Bruker). A total of 100 serum samples were examined, including 50 PDAC cases and 50 controls (25 females and 25 males in ea	CROSS-LABORATORY VALIDATION; GLYCOSPHINGOLIPIDS; PROFILING PDAC HIGH-RESOLUTION; ION MOBILITY; MASS SPECTROMETRY