%0 Journal Article
%A Li, Mujia Jenny
%A Meyer, Larissa Chiara
%A Meier, Nadine
%A Witte, Jannik
%A Maldacker, Maximilian
%A Seredynska, Adrianna
%A Schueler, Julia
%A Schilling, Oliver
%A Föll, Melanie Christine
%T Spatial Proteomics by Parallel Accumulation-Serial Fragmentation Supported MALDI MS/MS Imaging: A First Glance Into Multiplexed and Spatial Peptide Identification.
%J Rapid communications in mass spectrometry
%V 39
%N 9
%@ 0951-4198
%C New York, NY
%I Wiley Interscience
%M DKFZ-2025-00301
%P e10006
%D 2025
%X In spatial proteomics, matrix-assisted laser desorption/ionization (MALDI) imaging enables rapid and cost-effective peptide measurements. Yet, in situ peptide identification remains challenging. Therefore, this study aims to integrate the trapped ion mobility spectrometry (TIMS)-based parallel accumulation-serial fragmentation (PASEF) into MALDI imaging of tryptic peptides to enable multiplexed MS/MS imaging.An initial MALDI TIMS MS1 survey measurement was performed, followed by a manual generation of a precursor list containing mass over charge values and ion mobility windows. Inside the dual TIMS system, submitted precursors were trapped, separately eluted by their ion mobility and analyzed in a quadrupole time-of-flight device, thereby enabling multiplexed MALDI MS/MS imaging. Finally, precursors were identified by peptide to spectrum matching.This study presents the first multiplexed MALDI TIMS MS/MS imaging (iprm-PASEF) of tryptic peptides. Its applicability was showcased on two histomorphologically distinct tissue specimens in a four-plex and five-plex setup. Precursors were successfully identified by the search engine MASCOT in one single MALDI imaging experiment for each respective tissue. Peptide identifications were corroborated by liquid-chromatography tandem mass spectrometry experiments and fragment colocalization analyses.In this study, we present a novel pipeline, based on iprm-PASEF, that allows the multiplexed and spatial identification of tryptic peptides in MALDI imaging. Hence, it marks a first step towards the integration of MALDI imaging into the emerging field of spatial proteomics.
%K Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization: methods
%K Proteomics: methods
%K Tandem Mass Spectrometry: methods
%K Animals
%K Peptides: chemistry
%K Peptides: analysis
%K Ion Mobility Spectrometry: methods
%K Peptide Fragments: analysis
%K Peptide Fragments: chemistry
%K MALDI imaging (Other)
%K MS/MS imaging (Other)
%K TIMS (Other)
%K iprm‐PASEF (Other)
%K peptide imaging (Other)
%K spatial proteomics (Other)
%K Peptides (NLM Chemicals)
%K Peptide Fragments (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:39910729
%R 10.1002/rcm.10006
%U https://inrepo02.dkfz.de/record/298584