Determining Plasma Protein Variation Parameters as a Prerequisite for Biomarker Studies-A TMT-Based LC-MSMS Proteome Investigation.
Determining Plasma Protein Variation Parameters as a Prerequisite for Biomarker Studies-A TMT-Based LC-MSMS Proteome Investigation.
复制标题
DOI：
10.3390/proteomes9040047
复制
发表时间：
2021-12-01
期刊：
影响因子：
3.3
通讯作者：
Beck HC
中科院分区：
文献类型：
--
作者：
Andersen LC;Palstrøm NB;Diederichsen A;Lindholt JS;Rasmussen LM;Beck HC
特定的血浆蛋白质作为各种疾病的有价值的标志物，并且在许多情况下在临床实验室中通过全自动系统进行常规测量。为了使用这些标记物进行安全的诊断和监测，重要的是确保符合临床需求的分析质量。为此目的，在发现和验证新的疾病蛋白质生物标志物的背景下，关于测量的血浆蛋白的分析和生物学变异的信息是重要的，特别是与临床研究中的样本量计算有关。然而，关于大多数中高丰度血浆蛋白的生物学变异的信息在很大程度上是缺乏的。在这项研究中，我们假设，它是可能的，以产生数据的个体间的生物学变异结合分析变异的几百个丰富的血浆蛋白，通过应用LC-MS/MS结合相对定量使用同量异位素标记（10重TMT标记）的血浆样品。使用这种分析蛋白质组学方法，我们分析了42个血浆样本制备的双重，并估计技术，个体间生物，和总变异的265个最丰富的蛋白质存在于人血浆中，从而创造了功率分析和样本量确定在未来的临床蛋白质组学研究的先决条件。我们的结果表明，如果预期丰度的相对变化>1.5倍，则每组仅五个样品可以为大多数分析的蛋白质提供足够的统计功效。17种测量的蛋白质存在于欧洲临床化学和实验室医学联合会（EFLM）生物变异数据库中，并证明与EFLM数据库中列出的相应CV非常相似的生物CV，这表明生成的蛋白质组学确定的变异知识可用于大规模测定血浆蛋白变异。
Specific plasma proteins serve as valuable markers for various diseases and are in many cases routinely measured in clinical laboratories by fully automated systems. For safe diagnostics and monitoring using these markers, it is important to ensure an analytical quality in line with clinical needs. For this purpose, information on the analytical and the biological variation of the measured plasma protein, also in the context of the discovery and validation of novel, disease protein biomarkers, is important, particularly in relation to for sample size calculations in clinical studies. Nevertheless, information on the biological variation of the majority of medium-to-high abundant plasma proteins is largely absent. In this study, we hypothesized that it is possible to generate data on inter-individual biological variation in combination with analytical variation of several hundred abundant plasma proteins, by applying LC-MS/MS in combination with relative quantification using isobaric tagging (10-plex TMT-labeling) to plasma samples. Using this analytical proteomic approach, we analyzed 42 plasma samples prepared in doublets, and estimated the technical, inter-individual biological, and total variation of 265 of the most abundant proteins present in human plasma thereby creating the prerequisites for power analysis and sample size determination in future clinical proteomics studies. Our results demonstrated that only five samples per group may provide sufficient statistical power for most of the analyzed proteins if relative changes in abundances >1.5-fold are expected. Seventeen of the measured proteins are present in the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Biological Variation Database, and demonstrated remarkably similar biological CV’s to the corresponding CV’s listed in the EFLM database suggesting that the generated proteomic determined variation knowledge is useful for large-scale determination of plasma protein variations.