PXD048565

PXD048565 is an original dataset announced via ProteomeXchange.

Title	Open-Tubular Trap Columns: Towards Simple and Robust Liquid Chromatography Separations for Single-Cell Proteomics
Description	Nanoflow liquid chromatography-mass spectrometry is key to enabling in-depth proteome profiling of trace samples such as single cells, but these separations can lack robustness due to the use of narrow-bore columns that are susceptible to clogging. In the case of single-cell proteomics, offline cleanup steps are generally omitted to avoid losses to additional surfaces, and online solid-phase extraction/trap columns frequently provide the only opportunity to remove salts and insoluble debris before the sample is introduced to the analytical column. Trap columns are traditionally short, packed columns used to load and concentrate analytes at flow rates greater than those employed in analytical columns, and since these first encounter the uncleaned sample mixture, trap columns are also susceptible to clogging. We hypothesized that clogging could be avoided by using large-bore porous layer open tubular trap columns (PLOTrap). The low back pressure ensured that the PLOTraps could also serve as the sample loop, thus allowing sample cleanup and injection with a single 6-port valve. We found that PLOTraps could effectively remove debris to avoid column clogging. We also evaluated multiple stationary phases and PLOTrap diameters to optimize performance in terms of peak widths and sample loading capacities. Optimized PLOTraps were compared to conventional packed trap columns operated in forward and backflush modes, and were found to have similar chromatographic performance of backflushed traps while providing improved debris removal for robust analyses of trace samples.
HostingRepository	PRIDE
AnnounceDate	2024-02-09
AnnouncementXML	Submission_2024-02-09_09:04:34.955.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ryan Kelly
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-01-15 09:34:53	ID requested
⏵ 1	2024-02-09 09:04:35	announced

10.1039/D3MO00249G;

submitter keyword: solid-phase extraction, trap column, Open-tubular,Single-Cell proteomics

Ryan T. Kelly
contact affiliation	Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah, United States of America
contact email	ryan.kelly@byu.edu
lab head
Ryan Kelly
contact affiliation	Brigham Young University
contact email	ryan.kelly@byu.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/02/PXD048565

PRIDE project URI

• PRIDE
  1. PXD048565
     1. Label: PRIDE project
     2. Name: Open-Tubular Trap Columns: Towards Simple and Robust Liquid Chromatography Separations for Single-Cell Proteomics