Quantitative Analysis of the Intra- and Inter-Individual Variability of the Normal Urinary Proteome
Quantitative Analysis of the Intra- and Inter-Individual Variability of the Normal Urinary Proteome
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DOI：
10.1021/pr100835s
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发表时间：
2011-02-01
影响因子：
4.4
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
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作者：
Nagaraj, Nagarjuna;Mann, Matthias
尿液是一种容易且非侵入性获得的体液。基于质谱（MS）的蛋白质组学已经表明，尿液含有数千种蛋白质。尿液是近端和远端组织疾病的生物标志物的潜在来源，但它被认为比更常用的血浆更易变。通过LC-MS/MS分析的LTQ-Orbitrap没有prefractionation，我们的特点是尿蛋白质组的七个正常人捐助者在连续三天。一式三份单次运行的无标记定量涵盖了超过600种蛋白质的尿蛋白质组。技术重复的变异系数（cv）中位数为0.18。日间变异性明显较高，变异系数为0.48，个体间尿蛋白质组的总体变异为0.66。因此，我们的数据中的技术变异性为7.5%，而个人内部变异性贡献了45.5%，人际变异性贡献了47.1%。个体尿蛋白正常波动的测定在生物标志物研究中建立显著性阈值是有用的。我们的数据还允许定义一组常见的和丰富的500种蛋白质，这些蛋白质在所有研究的个体中都很容易检测到。该核心尿蛋白质组具有高比例的分泌、膜和相对高分子量蛋白质。
Urine is a readily and noninvasively obtainable body fluid. Mass spectrometry (MS)-based proteomics has shown that urine contains thousands of proteins. Urine is a potential source of biomarkers for diseases of proximal and distal tissues but it is thought to be more variable than the more commonly used plasma. By LC-MS/MS analysis on an LTQ-Orbitrap without prefractionation we characterized the urinary proteome of seven normal human donors over three consecutive days. Label-free quantification of triplicate single runs covered the urinary proteome to a depth of more than 600 proteins. The median coefficient of variation (cv) of technical replicates was 0.18. Interday variability was markedly higher with a cv of 0.48 and the overall variation of the urinary proteome between individuals was 0.66. Thus technical variability in our data was 7.5%, whereas intrapersonal variability contributed 45.5% and interpersonal variability contributed 47.1% to total variability. Determination of the normal fluctuation of individual urinary proteins should be useful in establishing significance thresholds in biomarker studies. Our data also allowed definition of a common and abundant set of 500 proteins that were readily detectable in all studied individuals. This core urinary proteome has a high proportion of secreted, membrane, and relatively high-molecular weight proteins.