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@ARTICLE{Distler:281523,
author = {Distler, Ute and Yoo, Han Byul and Kardell, Oliver and
Hein, Dana and Sielaff, Malte and Scherer, Marian and
Jozefowicz, Anna M and Leps, Christian and Gomez-Zepeda,
David and von Toerne, Christine and Merl-Pham, Juliane and
Barth, Teresa K and Tüshaus, Johanna and Giesbertz, Pieter
and Müller, Torsten and Kliewer, Georg and Aljakouch, Karim
and Helm, Barbara and Unger, Henry and Frey, Dario L and
Helm, Dominic and Schwarzmüller, Luisa and Popp, Oliver and
Qin, Di and Wudy, Susanne I and Sinn, Ludwig Roman and
Mergner, Julia and Ludwig, Christina and Imhof, Axel and
Kuster, Bernhard and Lichtenthaler, Stefan F and Krijgsveld,
Jeroen and Klingmüller, Ursula and Mertins, Philipp and
Coscia, Fabian and Ralser, Markus and Mülleder, Michael and
Hauck, Stefanie M and Tenzer, Stefan},
title = {{M}ulticenter evaluation of label-free quantification in
human plasma on a high dynamic range benchmark set.},
journal = {Nature Communications},
volume = {16},
number = {1},
issn = {2041-1723},
address = {[London]},
publisher = {Springer Nature},
reportid = {DZNE-2025-01141},
pages = {8774},
year = {2025},
abstract = {Human plasma is routinely collected during clinical care
and constitutes a rich source of biomarkers for diagnostics
and patient stratification. Liquid chromatography-mass
spectrometry (LC-MS)-based proteomics is a key method for
plasma biomarker discovery, but the high dynamic range of
plasma proteins poses significant challenges for MS analysis
and data processing. To benchmark the quantitative
performance of neat plasma analysis, we introduce a
multispecies sample set based on a human tryptic plasma
digest containing varying low level spike-ins of yeast and
E. coli tryptic proteome digests, termed PYE. By analysing
the sample set on state-of-the-art LC-MS platforms across
twelve different sites in data-dependent (DDA) and
data-independent acquisition (DIA) modes, we provide a data
resource comprising a total of 1116 individual LC-MS runs.
Centralized data analysis shows that DIA methods outperform
DDA-based approaches regarding identifications, data
completeness, accuracy, and precision. DIA achieves
excellent technical reproducibility, as demonstrated by
coefficients of variation (CVs) between $3.3\%$ and $9.8\%$
at protein level. Comparative analysis of different setups
clearly shows a high overlap in identified proteins and
proves that accurate and precise quantitative measurements
are feasible across multiple sites, even in a complex matrix
such as plasma, using state-of-the-art instrumentation. The
collected dataset, including the PYE sample set and strategy
presented, serves as a valuable resource for optimizing the
accuracy and reproducibility of LC-MS and bioinformatic
workflows for clinical plasma proteome analysis.},
keywords = {Humans / Proteomics: methods / Chromatography, Liquid:
methods / Blood Proteins: analysis / Blood Proteins:
metabolism / Proteome: analysis / Benchmarking /
Reproducibility of Results / Mass Spectrometry: methods /
Biomarkers: blood / Escherichia coli: metabolism / Plasma:
chemistry / Blood Proteins (NLM Chemicals) / Proteome (NLM
Chemicals) / Biomarkers (NLM Chemicals)},
cin = {AG Lichtenthaler},
ddc = {500},
cid = {I:(DE-2719)1110006},
pnm = {352 - Disease Mechanisms (POF4-352)},
pid = {G:(DE-HGF)POF4-352},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:41038884},
pmc = {pmc:PMC12491457},
doi = {10.1038/s41467-025-64501-z},
url = {https://pub.dzne.de/record/281523},
}