Efficient exploitation of separation space in two-dimensional liquid chromatography system for comprehensive and efficient proteomic analyses

Hangyeore Lee, Dong Gi Mun, Jeong Eun So, Jingi Bae, Hokeun Kim, Christophe Masselon, Sang Won Lee

    Research output: Contribution to journalArticlepeer-review

    16 Citations (Scopus)

    Abstract

    Proteomics aims to achieve complete profiling of the protein content and protein modifications in cells, tissues, and biofluids and to quantitatively determine changes in their abundances. This information serves to elucidate cellular processes and signaling pathways and to identify candidate protein biomarkers and/or therapeutic targets. Analyses must therefore be both comprehensive and efficient. Here, we present a novel online two-dimensional reverse-phase/reverse-phase liquid chromatography separation platform, which is based on a newly developed online noncontiguous fractionating and concatenating device (NCFC fractionator). In bottom-up proteomics analyses of a complex proteome, this system provided significantly improved exploitation of the separation space of the two RPs, considerably increasing the numbers of peptides identified compared to a conti uous 2D-RP/RPLC method. The fully automated online 2D-NCFC-RP/RPLC system bypassed a number of labor-intensive manual processes required with the previously described offline 2D-NCFC RP/RPLC method, and thus, it offers minimal sample loss in a context of highly reproducible 2D-RP/RPLC experiments.

    Original languageEnglish
    Pages (from-to)11734-11741
    Number of pages8
    JournalAnalytical chemistry
    Volume88
    Issue number23
    DOIs
    Publication statusPublished - 2016 Dec 6

    Bibliographical note

    Funding Information:
    This work was supported by the Multiomics Research Program (NRF-2012M3A9B9036675), Brain Research Program (NRF-2014M3C7A1046047), and International Research & Development Program (NRF-2015K1A3A1A21000273) funded by the Korean Ministry of Science, ICT & Future Planning. S.L. acknowledges LG Yonam Foundation.

    Publisher Copyright:
    © 2016 American Chemical Society.

    ASJC Scopus subject areas

    • Analytical Chemistry

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