通过nLC-MRM / MS在人血浆中定量分析87种蛋白质：用于生物样本库的大规模分析的工作流程,Journal of Proteome Research

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通过nLC-MRM / MS在人血浆中定量分析87种蛋白质：用于生物样本库的大规模分析的工作流程
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2017-08-08 00:00:00 , DOI: 10.1021/acs.jproteome.7b00235
Melinda Rezeli ₁ , Karin Sjödin ₂ , Henrik Lindberg ₁ , Olof Gidlöf ₃ , Bertil Lindahl ₄ , Tomas Jernberg ₅ , Jonas Spaak ₅ , David Erlinge ₃ , György Marko-Varga ₁

Affiliation

开发了一种多反应监测（MRM）分析方法，可使用纳米级液相色谱分离和稳定的同位素稀释策略对87种血浆蛋白进行精确定量，包括与心血管疾病相关的三种载脂蛋白E（APOE）亚型。对测定的分析性能进行了评估，我们发现整个血浆消化液在4-5个数量级的动态范围内（约0.2 mg / L至4.5 g / L），平均技术差异为4.7％。在这里，我们报告了完整的工作流程，包括适用于96孔板格式的样品处理以及用于大规模研究的归一化策略。为了进一步研究基于MS的定量，还通过常规使用的临床化学测定法测量了六种选定蛋白质的量，并且这两种方法显示出对这六种蛋白质的高度相关性（p值<10e-5），具有极好的相关性。对于心血管预测因子，APOB：APOA1比率（r = 0.969，p值<10e-5）。此外，我们利用发达的检测方法筛查了患有心肌梗塞的患者的生物库样品，并对STEMI（ST段抬高型心肌梗塞），NSTEMI（非ST段抬高型心肌梗塞）和2型患者进行了比较分析。 AMI（2型心肌梗塞）患者。

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Quantitation of 87 Proteins by nLC-MRM/MS in Human Plasma: Workflow for Large-Scale Analysis of Biobank Samples

A multiple reaction monitoring (MRM) assay was developed for precise quantitation of 87 plasma proteins including the three isoforms of apolipoprotein E (APOE) associated with cardiovascular diseases using nanoscale liquid chromatography separation and stable isotope dilution strategy. The analytical performance of the assay was evaluated and we found an average technical variation of 4.7% in 4–5 orders of magnitude dynamic range (≈0.2 mg/L to 4.5 g/L) from whole plasma digest. Here, we report a complete workflow, including sample processing adapted to 96-well plate format and normalization strategy for large-scale studies. To further investigate the MS-based quantitation the amount of six selected proteins was measured by routinely used clinical chemistry assays as well and the two methods showed excellent correlation with high significance (p-value < 10e-5) for the six proteins, in addition for the cardiovascular predictor factor, APOB: APOA1 ratio (r = 0.969, p-value < 10e-5). Moreover, we utilized the developed assay for screening of biobank samples from patients with myocardial infarction and performed the comparative analysis of patient groups with STEMI (ST- segment elevation myocardial infarction), NSTEMI (non ST- segment elevation myocardial infarction) and type-2 AMI (type-2 myocardial infarction) patients.

更新日期：2017-08-09

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