PXD052735-2

PXD052735 is an original dataset announced via ProteomeXchange.

Title	Integrated MICROFASP method with CZE based fractionation technique and nanoRPLC-ESI-MS/MS for comprehensive proteomics analysis of submicrogram sample
Description	We report a loss-less two dimensional (2D) separation platform that integrated capillary zone electrophoresis (CZE) fractionation and nanoRPLC-ESI-MS/MS for comprehensive proteomics analysis of submicrogram sample. Protein digest was injected into the linear polyacrylamide coated capillary followed by CZE separation. The schemes to collecting the fractions were carefully optimized to maximize the protein coverage. The peptide fractions were directly eluted into the autosampler insert vials followed by nanoRPLC-ESI-MS/MS analysis without lyophilization and redissolution, thus dramatically minimized sample loss and potential contamination. The integrated platform generated 30,845 unique peptides and 5,231 protein groups from 500 ng of a HeLa protein digest within 11.5 hours (90 min CZE fractionation plus 10 hours LC-MS analysis). Finally, the developed platform was used to analysis the protein digest prepared by MICROFASP method with 1 µg cell lysate as starting material. 3796 (N=2, RSD=4.95%) protein groups and 20,577(N=2, RSD=7.89%) peptides were identified from only 200 ng of the resulted tryptic digest within 5.5 hours. The results indicated that combination of MICROFASP method and the developed CZE/nanoRPLC-MS/MS 2D separation platform enabled comprehensive proteome profiling of submicrogram biological sample.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_06:54:36.921.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Zhenbin Zhang
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-05-30 19:20:07	ID requested
1	2024-08-10 16:16:23	announced
⏵ 2	2024-10-22 06:54:41	announced	2024-10-22: Updated project metadata.

10.1021/acs.jproteome.4c00396;

Lu G, Ying G, He Y, Li Y, Zhang Z, Integrated MICROFASP Method with CZE-Based Fractionation Technique and NanoRPLC-ESI-MS/MS for a Comprehensive Proteomics Analysis of a Submicrogram Sample. J Proteome Res, 23(8):3696-3703(2024) [pubmed]

submitter keyword: submicrogram sample, two dimensional separation, capillary zone electrophoresis, MICROFASP method,proteome profiling

Zhenbin Zhang
contact affiliation	Institute of Drug Discovery Technology, Ningbo University, Ningbo, Zhejiang, 315211 China
contact email	zhangzhenbin@nbu.edu.cn
lab head
Zhenbin Zhang
contact affiliation	Ningbo University
contact email	zhangzhenbin@nbu.edu.cn
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD052735
     1. Label: PRIDE project
     2. Name: Integrated MICROFASP method with CZE based fractionation technique and nanoRPLC-ESI-MS/MS for comprehensive proteomics analysis of submicrogram sample