オーラルセッション
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1A-O1-1650 PDF
四重極-Orbitrap ハイブリッド質量分析計を用いたハイスループットプロテオーム解析法の構築
Proteomics has become an increasingly important tool in medical and medicinal applications. It is necessary to improve the analytical throughput for these applications, particularly in large-scale drug screening to enable measurement of a large number of samples. In this study, we aimed to establish an ultrafast proteomic method based on a 5-min gradient LC and quadrupole-Orbitrap mass spectrometer (Q-Orbitrap MS). We precisely evaluated data-independent acquisition (DIA) parameters for 5-min gradient LC and reached a depth of >5,000 and 4,700 proteins from 1,000 and 250 ng HEK293T cell digest in a single-shot run, respectively. The throughput of our method enabled the measurement of approximately 80 samples/day, including sample loading, column equilibration, and wash running time. We demonstrated that our method is applicable for the screening of chemical responsivity via a cell stimulation assay. These data show that our method enables the capture of biological alterations in proteomic profiles with high sensitivity, suggesting the possibility of large-scale screening of chemical responsivity.