PXD028316

PXD028316 is an original dataset announced via ProteomeXchange.

Title	Middle-throughput LC-MS-based platelet proteomics with minute sample amounts using semi-automated positive pressure FASP in 384-well format (PF384)
Description	Comprehensive characterization of platelets involves various different functional assays and analysis techniques, each requiring individual aliquots of sample. Consequently, the available sample material is often limited rendering more effective sample use highly important. Our re-cently introduced proteomics workflow for semi-automated sample preparation, involving pos-itive pressure filter-aided sample preparation (FASP) in 96-well format (PF96), allows for highly robust platelet sample preparation but displays substantial analyte loss when processing minute protein amounts in the lower µg-range. Here we describe a downscaling of PF96 to 384-well format (PF384) in terms of required starting material. We determined meaningful sample loads and highlight advantages when processing only 3 µg purified platelet protein from 22 healthy donors, namely: (I) improved identification and analyte recovery with signal intensity gains of +130 % and +107 % (peptide and protein level, respectively) (II) substantial intensity gains for key-players in platelet activation including the membrane receptors PAR4, P2X1, GPVI, GPV, GPIX and the downstream mediators AKT, PKA, Rap1, Lyn (III) Reduction of variance when de-tecting disease markers by 6 %. Hence, these advantages, in conjunction with excellent through-put capabilities, render PF384 a promising future in clinical research and might even pave the way of platelet proteomics into molecular diagnostics.
HostingRepository	PRIDE
AnnounceDate	2025-02-14
AnnouncementXML	Submission_2025-02-14_05:04:49.297.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Stefan Loroch
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	carbamoylated residue; monohydroxylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-09-09 08:21:47	ID requested
⏵ 1	2025-02-14 05:04:49	announced

10.1055/A-2516-1812;

submitter keyword: 96-well format, High-Throughput, Platelet Signaling, PF384,Platelet Proteomics, PF96, Filter-Aided Sample Preparation, FASP, Automation, Platelet Disease Markers, 384 well format

Albert Sickmann
contact affiliation	Leibniz-Institut für Analytische Wissenschaften (ISAS), Dortmund, Germany
contact email	sickmann@isas.de
lab head
Stefan Loroch
contact affiliation	Medizinisches Proteome-Center der Ruhr Universität Bochum
contact email	stefan@loroch.email
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD028316
     1. Label: PRIDE project
     2. Name: Middle-throughput LC-MS-based platelet proteomics with minute sample amounts using semi-automated positive pressure FASP in 384-well format (PF384)