期刊
PROTEOMICS CLINICAL APPLICATIONS
卷 12, 期 2, 页码 -出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/prca.201700039
关键词
multiple reaction monitoring-mass spectrometry; N-15-labeled recombinant protein; quantitation; salivary proteins; variability assessment
资金
- Ministry of Education, Taiwan
- Ministry of Health and Welfare, Taiwan [PMRPD1B0102, PM-RPD1B0103, PMRPD1B0104]
- Ministry of Science and Technology [MOST 104-2325-B-182-003, MOST105-2325-B-182-001, MOST 103-2632-B-182-001, 104-2632-B-182-001, 105-2632-B-182-001]
- Chang Gung Memorial Hospital [CIRPD3B0013, CLRPD190017, CIRPD3E0013]
PurposeSaliva is an attractive sample source for the biomarker-based testing of several diseases, especially oral cancer. Here, we sought to apply multiplexed LC-MRM-MS to precisely quantify 90 disease-related proteins and assess their intra- and interindividual variability in saliva samples from healthy donors. Experimental designWe developed two multiplexed LC-MRM-MS assays for 122 surrogate peptides representing a set of disease-related proteins. Saliva samples were collected from 10 healthy volunteers at three different time points (Day 1 morning and afternoon, and Day 2 morning). Each sample was spiked with a constant amount of a N-15-labeled protein and analyzed by MRM-MS in triplicate. Quantitative results from LC-MRM-MS were calculated by single-point quantification with reference to a known amount of internal standard (heavy peptide). ResultsThe CVs for assay reproducibility and technical variation were 13 and 11%, respectively. The average concentrations of the 99 successfully quantified proteins ranged from 0.280.58ng mL(-1) for profilin-2 (PFN2) to 8.55 +/- 8.96 g mL(-1) for calprotectin (S100A8). For the 90 proteins detectable in >50% of samples, the average CVs for intraday, interday, intraindividual, and interindividual samples were 38%, 43%, 45%, and 69%, respectively. The fluctuations of most target proteins in individual subjects were found to be within +/- twofold. Conclusions and clinical relevanceOur study elucidated the intra- and interindividual variability of 90 disease-related proteins in saliva samples from healthy donors. The findings may facilitate the further development of salivary biomarkers for oral and systemic diseases.
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