| Summary: | 碩士 === 國立暨南國際大學 === 應用化學系 === 101 === Selective isolation and concentration of phosphopeptides is a prerequisite step prior to MS analysis. Currently titanium oxide affinity chromatography has gained more and more popularity among the common phosphopeptide isolation methods. Here, we present a new hybrid material, TiO2-coated nanodiamonds (TiO2-NDs), for selective enrichment of phosphopeptides from complex peptide mixtures. The results showed that TiO2-NDs outperformed the commercial TiO2 beads for higher enrichment capacity and specificity toward phosphopeptides. We identified 932 unique phosphopeptides and 643 phosphorylation site from 100 μg of HeLa cells, which represented a 1.5-fold increase in phosphopeptide number and a 1.5-fold increase in phosphosites in comparison with the results obtained using commercial TiO2 beads. This new hybrid material (TiO2-NDs) can be easily incorporated as an effective alternative for TiO2 beads into existing protocols for phosphopeptide isolation, and can facilitate more comprehensive profiling of phosphoproteome.
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