PXD053720

PXD053720 is an original dataset announced via ProteomeXchange.

Title	A fast, high-sensitivity 96-well plate based MICROFASP method for processing low microgram proteomics sample within 1.5 hours
Description	We report a fast, high-sensitivity MICROFASP method that capable of completing sample preparation in less than 1.5 h which improved the throughput over 13 times compared to the previous reports. In this method, protein digestion time was dramatically shortened from 17 h to 20 min in the limited reaction volume. To further decrease the sample preparation time, we used tris(2-carboxyethyl)phosphine and Iodoacetamide (IAA) for the reduction of disulfide bonds and the alkylation of the resulted free thiol groups simultaneously instead of using dithiothreitol and IAA which has to be carried out the reactions in two separate steps. Label free quantitation (LFQ) intensities of the proteins identified from the fast MICROFASP method and the conventional MICROFASP method were highly correlated (the pearson correlation coefficient r=0.907) which confirmed the reliability of the fast MICROFASP method. We also developed 96-well plate based fast-MICROFASP method to process 8 brain tissue samples from APP/PS1 transgenic mice in parallel. Amyloid-beta protein, a pathological protein of Alzheimer’s disease, was successfully identified as the high expressed protein in the brain of APP/PS1 transgenic mice, suggesting the diagnositic potential of our fast-MICROFASP method. We anticipate the fast, high-sensitivity 96-well plate based MICROFASP method has wide application in high-throughput and rapid preparation of large cohorts of low microgram samples (e.g., clinical biopsy) for comprehensive proteome profiling.
HostingRepository	PRIDE
AnnounceDate	2025-05-07
AnnouncementXML	Submission_2025-05-06_21:16:06.080.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Zhenbin Zhang
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; deaminated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-07-07 03:19:07	ID requested
⏵ 1	2025-05-06 21:16:06	announced

Ying G, He Y, Yang M, Lu G, Li Y, Cui W, Hu Z, Zhang Z, A Fast, High-Sensitivity 96-Well Plate-Based MICROFASP Method for Processing Low Microgram Proteomics Sample within 1.5 h. Anal Chem, 97(4):2111-2119(2025) [pubmed]

10.1021/acs.analchem.4c04857;

submitter keyword: fast sample preparation

proteomics analysis

high-sensitivity

high-throughput

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. PXD053720
     1. Label: PRIDE project
     2. Name: A fast, high-sensitivity 96-well plate based MICROFASP method for processing low microgram proteomics sample within 1.5 hours