Phenyl‐bonded monolithic silica capillary column liquid chromatographic separation and detection of fluorogenic derivatized intact proteins
作者： Hiroo Wada , Hiroshi Kobayashi , Kazuhiro Imai
DOI: 10.1002/BMC.5078
关键词: Yeast 、 Proteomics 、 Chromatography 、 Mass spectrum 、 Reagent 、 Proteome 、 Molar mass distribution 、 Mass spectrometry 、 High-performance liquid chromatography 、 Chemistry
摘要: Prior to the identification of proteins for proteomics analysis in human cells, separation fluorogenic derivatized with a reagent, 7-chloro-N-[2-(dimethylamino)ethyl]-2,1,3-benzoxadiazole-4-sulfonamide, has typically been performed by using conventional reversed-phase HPLC column. However, number cells (HepaRG) that are separated this approach is limited approximately 500. In study, nanoflow liquid chromatography system an evaluated phenyl-bonded monolithic silica capillary column (0.1 mm i.d., 700 length) was used increase proteins. This separate cell (K562) and yeast (Saccharomyces cerevisiae) as model proteomes. More than 1,300 protein peaks were separated/detected from both We present straightforward comparison multiple profiles novel chromatogram display approach, termed "spiderweb" chromatogram. addition, validate detected derived proteins, mass spectrometer connected deconvolution obtained spectra performed. Furthermore, different molecular weight distribution expressed observed between two
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