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PXD039290-1

PXD039290 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleTargeted quantitative mass spectrometry analysis of protein biomarkers from previously stained single formalin-fixed paraffin-embedded tissue sections
DescriptionFormalin-fixed, paraffin-embedded (FFPE) tissues represent a majority of all biopsy specimens commonly analyzed by histological or immunohistochemical (IHC) staining with adhesive coverslips attached. Mass spectrometry (MS) recently has been employed to precisely quantify proteins in samples consisting of multiple unstained FFPE sections. Here we report a MS method to analyze proteins from a single coverslipped 4-micron section previously stained with hematoxylin/eosin (H&E), Masson’s trichrome, or 3,3’-diaminobenzidine based IHC. We analyzed serial unstained and stained sections from non-small cell lung cancer (NSCLC) specimens for proteins of varying abundance (PD-L1, RB1, CD73, HLA-DRA). Coverslips were removed by soaking in xylene and, after tryptic digestion, peptides were analyzed by targeted high-resolution LC-MS/MS with stable isotope labeled peptide standards. The low abundance proteins RB1 and PD-L1 were quantified in 31 and 35 of 50 total sections analyzed, respectively, whereas higher abundance CD73 and HLA-DRA were quantified in 49 and 50 sections. Inclusion of targeted beta-actin (ACTB) measurement enabled normalization in samples where residual stain interfered with bulk protein quantitation by colorimetric assay. Measurement CVs for 5 replicate slides (H&E vs. unstained) from each block ranged from 3-18% for PD-L1, from 1-36% for RB1, from 3-21% for CD73 and from 4-29% for HLA-DRA. Collectively, these results demonstrate that targeted MS protein quantification can add a valuable data layer to clinical tissue specimens after assessment for standard pathology endpoints.
HostingRepositoryPanoramaPublic
AnnounceDate2026-06-12
AnnouncementXMLSubmission_2026-06-12_10:22:49.057.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterRyan Morrison
SpeciesList scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationListLabel:13C(6)15N(2); Label:13C(6)15N(4)
InstrumentOrbitrap Exploris 480
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-01-05 09:30:39ID requested
12026-06-12 10:22:51announced
Publication List
Ackermann BL, Morrison RD, Hill S, Westfall MD, Butts BD, Soper MD, Fill JA, Schade AE, Liebler DC, Gruver AM, Targeted Quantitative Mass Spectrometry Analysis of Protein Biomarkers From Previously Stained Single Formalin-Fixed Paraffin-Embedded Tissue Sections. Lab Invest, 103(4):100052(2023) [pubmed]
Keyword List
submitter keyword: Formalin-fixed tissue, mass spectrometry, quantitative proteomics, single section
Contact List
Daniel Liebler
contact affiliationProtypia, Inc
contact emaildaniel.liebler@protypia.com
lab head
Ryan Morrison
contact affiliationProtypia, Inc
contact emailryan.morrison@protypia.com
dataset submitter
Full Dataset Link List
Panorama Public dataset URI