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@ARTICLE{Wacker:278342,
author = {M. Wacker and J. Bauer and L. Wessling and M. Dubbelaar and
A. Nelde and H.-G. Rammensee$^*$ and J. S. Walz$^*$},
title = {{I}mmunoprecipitation methods impact the peptide repertoire
in immunopeptidomics.},
journal = {Frontiers in immunology},
volume = {14},
issn = {1664-3224},
address = {Lausanne},
publisher = {Frontiers Media},
reportid = {DKFZ-2023-01599},
pages = {1219720},
year = {2023},
abstract = {Mass spectrometry-based immunopeptidomics is the only
unbiased method to identify naturally presented HLA ligands,
which is an indispensable prerequisite for characterizing
novel tumor antigens for immunotherapeutic approaches. In
recent years, improvements based on devices and methodology
have been made to optimize sensitivity and throughput in
immunopeptidomics. However, developments in ligand
isolation, mass spectrometric analysis, and subsequent data
processing can have a marked impact on the quality and
quantity of immunopeptidomics data.In this work, we compared
the immunopeptidome composition in terms of peptide yields,
spectra quality, hydrophobicity, retention time, and
immunogenicity of two established immunoprecipitation
methods (column-based and 96-well-based) using cell lines as
well as primary solid and hematological tumor
samples.Although, we identified comparable overall peptide
yields, large proportions of method-exclusive peptides were
detected with significantly higher hydrophobicity for the
column-based method with potential implications for the
identification of immunogenic tumor antigens. We showed that
column preparation does not lose hydrophilic peptides in the
hydrophilic washing step. In contrast, an additional $50\%$
acetonitrile elution could partially regain lost hydrophobic
peptides during 96-well preparation, suggesting a reduction
of the bias towards the column-based method but not
completely equalizing it.Together, this work showed how
different immunoprecipitation methods and their adaptions
can impact the peptide repertoire of immunopeptidomic
analysis and therefore the identification of potential
tumor-associated antigens.},
keywords = {HLA peptides (Other) / hydrophobicity (Other) /
immunogenicity (Other) / immunopeptidomics (Other) /
immunoprecipitation (Other) / immunotherapies (Other) / mass
spectrometry (Other)},
cin = {TU01},
ddc = {610},
cid = {I:(DE-He78)TU01-20160331},
pnm = {899 - ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:37545538},
pmc = {pmc:PMC10400765},
doi = {10.3389/fimmu.2023.1219720},
url = {https://inrepo02.dkfz.de/record/278342},
}