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PXD003370

PXD003370 is an original dataset announced via ProteomeXchange.

Dataset Summary
Title1D LC-MS/MS proteomic profiling of MCF10A, HeLa, Hep G2, and HEK293 cells using peptide-based immunoaffinity enrichment and 105 commercially available monoclonal antibodies to proteins associated with the DNA damage response (DDR) network
DescriptionImmunoaffinity enrichment of peptides coupled to multiple reaction monitoring-mass spectrometry (immuno-MRM) enables highly specific, sensitive, and precise quantification of peptides and post-translational modifications. Major obstacles to developing a large number of immuno-MRM assays are the poor availability of monoclonal antibodies (mAbs) validated for immunoaffinity enrichment of peptides and the cost and lead time of developing the antibodies de novo. Although many thousands of mAbs are commercially offered, few have been tested for application to immunoaffinity enrichment of peptides. In this study we tested the success rate of using commercially available mAbs for peptide immuno-MRM assays. We selected 105 commercial mAbs (76 targeting non-modified “pan” epitopes, 29 targeting phosphorylation) to proteins associated with the DNA damage response network. We found that 8 of the 76 pan (11%) and 5 of the 29 phospho-specific mAbs (17%) captured tryptic peptides (detected by LC-MS/MS) of their protein targets from human cell lysates. Seven of these mAbs were successfully used to configure and analytically characterize immuno-MRM assays. By applying selection criteria, the results indicate a success rate up to 24% is possible, establishing the feasibility of screening a large number of catalog antibodies to provide readily-available assay reagents.
HostingRepositoryPRIDE
AnnounceDate2016-04-13
AnnouncementXMLSubmission_2016-04-13_02:10:20.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterJacob Kennedy
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListcarbamoylated residue; phosphorylated residue
InstrumentLTQ Orbitrap Velos
Dataset History
RevisionDatetimeStatusChangeLog Entry
02015-12-17 23:42:54ID requested
12016-04-13 02:10:21announced
Publication List
Dataset with its publication pending
Keyword List
ProteomeXchange project tag: CPTAC Consortium
curator keyword: Biomedical
submitter keyword: Human, monoclonal antibodies, MCF10A, HeLa, HEK293, Hep G2, IR, LC-MS/MS
Contact List
Amanda G Paulovich
contact affiliationFred Hutchinson Cancer Research Center
contact emailapaulovi@fredhutch.org
lab head
Jacob Kennedy
contact affiliationFred Hutchinson Cancer Research Center
contact emailjkennedy@fhcrc.org
dataset submitter
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Dataset FTP location
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