PXD049248

PXD049248 is an original dataset announced via ProteomeXchange.

Title	Multicolumn Nanoflow Liquid Chromatography System with Accelerated Offline Gradient Generation for Robust and Sensitive Single-Cell Proteome Profiling
Description	Peptide separations that combine high sensitivity, robustness, peak capacity and throughput are essential for extending bottom-up proteomics to smaller samples including single cells. To this end, we have developed a multicolumn nanoLC system with accelerated offline gradient generation. One binary pump generates gradients in an accelerated manner to support multiple analytical columns, and a single trap column interfaces with all analytical columns to reduce required maintenance and simplify troubleshooting. A high degree of parallelization is possible, as one sample undergoes separation while the next sample plus its corresponding mobile phase gradient are transferred into the storage loop and a third sample is loaded into a sample loop. Selective offline elution from the trap column into the sample loop prevents salts and hydrophobic species from entering the analytical column, thus greatly enhancing column lifetime and system robustness. With this design, samples can be analyzed as fast as every 20 min at a flow rate of just 40 nL/min with close to 100% MS utilization time, and continuously for as long as several months without column replacement. We utilized the system to analyze the proteomes of single cells from a multiple myeloma cell line upon treatment with the immunomodulatory imide drug lenalidomide.
HostingRepository	PRIDE
AnnounceDate	2025-05-06
AnnouncementXML	Submission_2025-05-06_13:40:18.887.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	2024-02-07 09:36:06	ID requested
⏵ 1	2025-05-06 13:40:19	announced

Xie X, Truong T, Huang S, Johnston SM, Hovanski S, Robinson A, Webber KGI, Lin HL, Mun DG, Pandey A, Kelly RT, Multicolumn Nanoflow Liquid Chromatography with Accelerated Offline Gradient Generation for Robust and Sensitive Single-Cell Proteome Profiling. Anal Chem, 96(26):10534-10542(2024) [pubmed]

10.1021/acs.analchem.4c00878;

submitter keyword: High throughput, proteomics, LC-MS, storage loop, single cell proteomics, multiple columns

Ryan T. Kelly
contact affiliation	Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 84602
contact email	ryan.kelly@micromicstech.com
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/2025/05/PXD049248

PRIDE project URI

• PRIDE
  1. PXD049248
     1. Label: PRIDE project
     2. Name: Multicolumn Nanoflow Liquid Chromatography System with Accelerated Offline Gradient Generation for Robust and Sensitive Single-Cell Proteome Profiling