Accurate Quantification of Cardiovascular Biomarkers in Serum Using Protein Standard Absolute Quantification (PSAQ™) and Selected Reaction Monitoring
Accurate Quantification of Cardiovascular Biomarkers in Serum Using Protein Standard Absolute Quantification (PSAQ™) and Selected Reaction Monitoring
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DOI：
10.1074/mcp.m111.008235
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发表时间：
2012-02-01
影响因子：
7
通讯作者：
Brun, Virginie
中科院分区：
文献类型：
--
作者：
Huillet, Celine;Adrait, Annie;Brun, Virginie
新的生物标志物的开发需要大大加快，以改善诊断，预后和毒性监测以及治疗随访。生物标志物评价是这一发展过程中的主要瓶颈。选择反应监测（SRM）结合稳定同位素稀释已成为一个有前途的选择，以加快这一步，特别是因为它的多路复用能力。然而，由于上游样品处理或不完全胰蛋白酶消化而导致的分析变异性仍需要解决。2007年，我们开发了PSAQ（TM）方法（蛋白质标准品绝对定量），该方法使用全长同位素标记的蛋白质标准品来定量靶蛋白。在本研究中，我们使用临床验证的心血管生物标志物（LDH-B、CKMB、肌红蛋白和肌钙蛋白I）来证明PSAQ和SRM（PSAQ-SRM）的组合可以高度准确地定量血清样本中的生物标志物。多重PSAQ-SRM测定用于定量来自心肌梗死患者的临床样品中的这些生物标志物。发现PSAQ-SRM和ELISA测定结果之间具有良好的相关性，并证明了这些分析方法之间的一致性。因此，PSAQ-SRM有能力提高蛋白质分析的准确性和重现性。这将是对有效的生物标志物开发策略的重大贡献。Molecular & Cellular Proteomics 11：10.1074/mcp. M111.008235，1-12，2012.
Development of new biomarkers needs to be significantly accelerated to improve diagnostic, prognostic, and toxicity monitoring as well as therapeutic follow-up. Biomarker evaluation is the main bottleneck in this development process. Selected Reaction Monitoring (SRM) combined with stable isotope dilution has emerged as a promising option to speed this step, particularly because of its multiplexing capacities. However, analytical variabilities because of upstream sample handling or incomplete trypsin digestion still need to be resolved. In 2007, we developed the PSAQ (TM) method (Protein Standard Absolute Quantification), which uses full-length isotope-labeled protein standards to quantify target proteins. In the present study we used clinically validated cardiovascular biomarkers (LDH-B, CKMB, myoglobin, and troponin I) to demonstrate that the combination of PSAQ and SRM (PSAQ-SRM) allows highly accurate biomarker quantification in serum samples. A multiplex PSAQ-SRM assay was used to quantify these biomarkers in clinical samples from myocardial infarction patients. Good correlation between PSAQ-SRM and ELISA assay results was found and demonstrated the consistency between these analytical approaches. Thus, PSAQ-SRM has the capacity to improve both accuracy and reproducibility in protein analysis. This will be a major contribution to efficient biomarker development strategies. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.008235, 1-12, 2012.