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    High-throughput microbore ultrahigh-performance liquid chromatography-ion mobility-enabled-mass spectrometry-based proteomics methodology for the exploratory analysis of serum samples from large cohort studies

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    Authors
    Lennon, S.
    Hughes, C. J.
    Muazzam, Ammara
    Townsend, Paul A
    Gethings, L. A.
    Wilson, I. D.
    Plumb, R. S.
    Affiliation
    Waters Corporation, Stamford Avenue, Wilmslow SK9 4AX, U.K.
    Issue Date
    2021
    
    Metadata
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    Abstract
    The deployment of proteomic analysis in clinical studies represents a significant opportunity to detect and validate biomarkers in translational medicine, improve disease understanding, and provide baseline information on population health. However, comprehensive proteome studies usually employ nanoscale chromatography and often require several hours of analysis/sample. Here, we describe a high-throughput liquid chromatography tandem mass spectrometry (LC/MS/MS) methodology using 1 mm scale chromatography requiring only 15 min/sample, coupled to ion mobility-enabled mass spectrometry. The short run time effected a 6-fold increase in productivity compared with nanoscale LC/MS. The method demonstrated excellent reproducibility with retention time coefficient of variations of less than 0.05% and peak area reproducibility ranging from 5 to 15%. The 1 mm system produced similar chromatographic peak capacity values to the nanoscale miniaturized system, detecting 90% of the Escherichia coli proteins identified by the 75 μm LC/MS system (albeit based on only 75% of the peptides found by the latter). Application to the analysis of serum samples from a human prostate cancer study group resulted in the identification of a total of 533 proteins revealing the differential expression of proteins linked to patients receiving hormone-radiotherapy or undergoing surgery.
    Citation
    Lennon S, Hughes CJ, Muazzam A, Townsend PA, Gethings LA, Wilson ID, et al. High-Throughput Microbore Ultrahigh-Performance Liquid Chromatography-Ion Mobility-Enabled-Mass Spectrometry-Based Proteomics Methodology for the Exploratory Analysis of Serum Samples from Large Cohort Studies. J Proteome Res. 2021;20(3):1705-15.
    Journal
    Journal of Proteome Research
    URI
    http://hdl.handle.net/10541/623843
    DOI
    10.1021/acs.jproteome.0c00821
    PubMed ID
    33566619
    Additional Links
    https://dx.doi.org/10.1021/acs.jproteome.0c00821
    Type
    Article
    Language
    en
    ae974a485f413a2113503eed53cd6c53
    10.1021/acs.jproteome.0c00821
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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