JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST001513/files/C1_LPA_1uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C3_Sph_1uM_03.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_10uM_03.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_01uM_01.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_10uM_01.rawhttps://storage.jpostdb.org/JPST001513/files/C3_Sph_1uM_01.rawhttps://storage.jpostdb.org/JPST001513/files/C3_Sph_01uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_1uM_03.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_01uM_03.rawhttps://storage.jpostdb.org/JPST001513/files/C1_LPA_10uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C1_LPA_1uM_03.rawhttps://storage.jpostdb.org/JPST001513/files/C1_LPA_10uM_01.rawhttps://storage.jpostdb.org/JPST001513/files/C3_Sph_1uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_10uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C3_Sph_01uM_03.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_1uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_01uM_02.rawhttps://storage.jpostdb.org/JPST001513/files/C2_S1P_1uM_01.rawhttps://storage.jpostdb.org/JPST001513/files/C3_Sph_01uM_01.rawhttps://storage.jpostdb.org/JPST001513/files/C1_LPA_10uM_03.rawprimaryOK200ProteomicsYusuke KawashimaHomo Sapiens (human)https://repository.jpostdb.org/entry/JPST001513Kazusa DNA InstitutejPOSTProteomics 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 5-min gradient LC and quadrupole-Orbitrap mass spectrometer (Q-Orbitrap MS). We precisely optimized data-independent acquisition (DIA) parameters for 5-min gradient LC and reached a depth of >5000 and 4200 proteins from 1000 and 31.25 ng of 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.Optimization of Ultrafast Proteomics Using an LC-Quadrupole-Orbitrap Mass Spectrometer with Data-Independent Acquisition.Ishikawa Masaki M, Konno Ryo R, Nakajima Daisuke D, Gotoh Mari M, Fukasawa Keiko K, Sato Hironori H, Nakamura Ren R, Ohara Osamu O, Kawashima Yusuke YHEK293T, Hek 293T, Human Embryonic Kidney 293T, 293-T, HEK-293T, 293tsA1609neo, ATP:Fas-activated serine/threonine protein phosphotransferase activity, determination, chemical analysis, STK10, total expressed protein, Cell., HEK 293 T, assay, FASTK, Proteomes, 293 T, FAST, Fast, HEK-293-T, 293Tl(2)k04204, 293 T cell, scale tissue, CDF, Product, determination, leg, Mbp1, 6330543G17Rik, Long Term, l(1)AA33, dmTAF[[II]]230, Techniques, POF, Method, responsivity, Pharmaceutical Product, 1, 2, Min, AGAMOUS-like 61, myd, Effect, DmelCG42273, DiA, HEK293T, l(2)k07135, TFIID TAF250, cel, DmelCG13176, plant peltate hair, Su(Mir)1, min, Mbp-1, ORF19, mAPC, DIASP, DIA, Dia, AI047805, Hek 293T, allergic reaction, 4-(4-dihexadecylaminostyryl)N-methylpyridium iodide, Methodological Studies, column, l(1)B2/13.1, medicine, RUN, Run, Pharmaceutical, scientific observation, sample, 38E.16, l(2)CA4, Prx3, Long-Term Effects, dTAF[[II]]230, l(1)LB9, HEK293T cell, gyltl1b-b, Dmel_CG7826, Longterm Effect, TAF200, Procedure, TAFII-250, TAF250/230, AA33, shortstop/kakapo, ms(2)04138, Desmoplastic infantile astrocytoma, TAFII250, Dm1, MDDGA6, mKIAA0609, DmelCG18076, Pharmaceutic, 1110049F14Rik, WASH, KIAA0609, Dmel_CG7835, Mnb, MNB, fg, gyltl1b, Grv, POF2, F27C12_24, mdc1d, F27C12.24, expanded, Methodological, AW124434, CG17603, Methodological Study, 293 T, TAF[[II]], SHOT, 293T cell, MDC1D, enr, enlarged, Taf250, Specificity and Sensitivity, grv, SR3-5, DIANA, 42/4, TAF230, 293-T cell, big, CG1849, OG12X, d230, Procedures, 293tsA1609neo, Peptidomics, Effects, peltate hair, number, Gene, dTAFII250, EfW1, froggy, CG7826, Gyltl1a, 293T, l(2)k05821, large, method, l(2)k15606, dmTAF1, Taf230, sensitive, p63, method used in an experiment, DmelCG1768, p65, Studies, CG7835, HILDA, Gene Products, CG42273, Technique, sensitivity, Shot, TAF250, Drugs, study, reactivity, Taf200, l(2)k10821, FAM39E, dTAF[[II]]250, 293-T, Longterm, cell, MDDGB6, Runt, Taf1p, LARGE, Long-Term, Study, BPFD#36, dTAF250, drugs, HEK-293T, great, Sensitivity, Long-Term Effect, Preparation, DIA2, TAF, LB5, Pharmaceuticals, Products, measuring, Human Embryonic Kidney 293T, DYRK1, TAF[[II]]250, kop, drug, Desmoplastic astrocytoma of infancy, Proteins, DmelCG1849, CG18076, HEK 293 T, l(3)84Ab, responsivity., BG:DS00004.13, CG1768, CG18637, Rnt, DRF2, Cell, dTAF230, lLB5, Wash2, Wash1, p230, MLPLI, Protein, Long Term Effects, chemical analysis, TAF[[II]]250/230, TFIID, Dyrk1, scales, Dias, CG13176, AU020952, Taf[[II]]250, l(2)k05434, TAF[[II]]230, scale, Pharmaceutic Preparations, l(2)k03010, CC1, Specificity, kak, l(1)19Ea, TAF[II]250, OG12, sample population, Longterm Effects, ME-IV, Drug, plan specification, Protein Gene Products, P235, Gene Proteins, Preparations, DmelCG17603, cardinality, assay, response, Og12x, Pharmaceutical Products, Runnings, HEK-293-T, Pharmaceutical Preparation, TAF1DIA., PeptidomicsMnb, MNB, AU020952, DYRK1, Procedures, CC1, Dmel_CG7826, min, mAPC, Methodological, Procedure, AW124434, Methodological Study, CG7826, plan specification, AI047805, Study, method, Techniques, Methodological Studies, Dm1, Method, ME-IV., method used in an experiment, Studies, CG7835, CG42273, Min, Dyrk1, Technique, Dmel_CG7835, DmelCG42273falseUltra-fast proteome analysis of HEK293T cells using Q-Orbitrap MSWe established an ultrafast proteomic method based on a 5-min gradient LC and quadrupole-Orbitrap mass spectrometer.Thu Jul 28 00:00:00 BST 2022PXD034227960635914019