The Johns Hopkins Bayview Proteomics Center, Johns Hopkins University, Baltimore, MD 21224, USA.
Proteomics Clin Appl. 2010 Mar;4(3):271-84. doi: 10.1002/prca.200900217. Epub 2010 Jan 7.
Over the last decade, translational science has come into the focus of academic medicine, and significant intellectual and financial efforts have been made to initiate a multitude of bench-to-bedside projects. The quest for suitable biomarkers that will significantly change clinical practice has become one of the biggest challenges in translational medicine. Quantitative measurement of proteins is a critical step in biomarker discovery. Assessing a large number of potential protein biomarkers in a statistically significant number of samples and controls still constitutes a major technical hurdle. Multiplexed analysis offers significant advantages regarding time, reagent cost, sample requirements and the amount of data that can be generated. The two contemporary approaches in multiplexed and quantitative biomarker validation, antibody-based immunoassays and MS-based multiple (or selected) reaction monitoring, are based on different assay principles and instrument requirements. Both approaches have their own advantages and disadvantages and therefore have complementary roles in the multi-staged biomarker verification and validation process. In this review, we discuss quantitative immunoassay and multiple reaction monitoring/selected reaction monitoring assay principles and development. We also discuss choosing an appropriate platform, judging the performance of assays, obtaining reliable, quantitative results for translational research and clinical applications in the biomarker field.
在过去的十年中，转化科学已经成为学术医学的焦点，并且已经投入了大量的智力和财力来启动众多从实验室到病床的项目。寻找能够显著改变临床实践的合适生物标志物已成为转化医学中最大的挑战之一。蛋白质的定量测量是生物标志物发现的关键步骤。在大量的样本和对照中对大量潜在的蛋白质生物标志物进行统计上显著的评估仍然是一个主要的技术障碍。多重分析在时间、试剂成本、样品需求和可以产生的数据量方面具有显著优势。当前用于多重和定量生物标志物验证的两种方法，基于抗体的免疫测定法和基于 MS 的多重（或选择）反应监测，基于不同的检测原理和仪器要求。这两种方法都有各自的优缺点，因此在多阶段生物标志物验证和验证过程中具有互补作用。在这篇综述中，我们讨论了定量免疫测定和多重反应监测/选择反应监测测定原理和开发。我们还讨论了选择合适的平台、判断测定的性能、为转化研究和生物标志物领域的临床应用获得可靠、定量的结果。
