title: Efficient Exploitation of Separation Space in Two-Dimensional Liquid Chromatography System for Comprehensive and Efficient Proteomic Analyses.
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 contiguous 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.
authors: Department of Chemistry, Research Institute for Natural Sciences, Korea University , Seoul 136-701, South Korea.
authors: Hangyeore
authors: H
authors: Lee
authors: Department of Chemistry, Research Institute for Natural Sciences, Korea University , Seoul 136-701, South Korea.
authors: Dong-Gi
authors: DG
authors: Mun
authors: Department of Chemistry, Research Institute for Natural Sciences, Korea University , Seoul 136-701, South Korea.
authors: Jeong Eun
authors: JE
authors: So
authors: Department of Chemistry, Research Institute for Natural Sciences, Korea University , Seoul 136-701, South Korea.
authors: Jingi
authors: J
authors: Bae
authors: Department of Chemistry, Research Institute for Natural Sciences, Korea University , Seoul 136-701, South Korea.
authors: Hokeun
authors: H
authors: Kim
authors: Université Grenoble Alpes , F-38402 Saint-Martin-d'Heres, France.|CEA, Institut de Biosciences et de Biotechnologie de Grenoble, Biologie à Grande Echelle, F-38054 Grenoble, France.|INSERM, U1038, F-38054 Grenoble, France.
authors: Christophe
authors: C
authors: Masselon
authors: Department of Chemistry, Research Institute for Natural Sciences, Korea University , Seoul 136-701, South Korea.
authors: Sang-Won
authors: SW
authors: Lee
mesh_terms: D056148:Chromatography, Reverse-Phase / Q000295:instrumentation; D006801:Humans; D009862:Online Systems; D010455:Peptides / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation
chemical_list: D010455:Peptides