Thermo Fisher Scientific, Biomarker research in Mass Spectrometry, Cambridge, Massachusettes, United States.
J Proteome Res. 2010 Dec 3;9(12):6678-88. doi: 10.1021/pr100821m. Epub 2010 Nov 2.
Mass spectrometry (MS) is an attractive alternative to quantification of proteins by immunoassays, particularly for protein biomarkers of clinical relevance. Reliable quantification requires that the MS-based assays are robust, selective, and reproducible. Thus, the development of standardized protocols is essential to introduce MS into clinical research laboratories. The aim of this study was to establish a complete workflow for assessing the transferability and reproducibility of selected reaction monitoring (SRM) assays between clinical research laboratories. Four independent laboratories in North America, using identical triple-quadrupole mass spectrometers (Quantum Ultra, Thermo), were provided with standard protocols and instrumentation settings to analyze unknown samples and internal standards in a digested plasma matrix to quantify 51 peptides from 39 human proteins using a multiplexed SRM assay. The interlaboratory coefficient of variation (CV) was less than 10% for 25 of 39 peptides quantified (12 peptides were not quantified based upon hydrophobicity) and exhibited CVs less than 20% for the remaining peptides. In this report, we demonstrate that previously developed research platforms for SRM assays can be improved and optimized for deployment in clinical research environments.
质谱法 (MS) 是免疫测定法定量蛋白质的一种有吸引力的替代方法，特别是对于具有临床相关性的蛋白质生物标志物。可靠的定量需要基于 MS 的测定具有稳健性、选择性和重现性。因此，制定标准化方案对于将 MS 引入临床研究实验室至关重要。本研究的目的是建立一种完整的工作流程，用于评估选定反应监测 (SRM) 测定在临床研究实验室之间的可转移性和重现性。北美四个独立的实验室，使用相同的三重四极杆质谱仪 (Quantum Ultra、Thermo)，提供了标准的协议和仪器设置，用于分析未知样品和消化后的血浆基质中的内标，以使用多重 SRM 测定定量 39 个人蛋白中的 51 个肽。在定量的 39 个肽中的 25 个中，实验室间的变异系数 (CV) 小于 10%（12 个肽由于疏水性而未定量），其余肽的 CV 小于 20%。在本报告中，我们证明了先前开发的用于 SRM 测定的研究平台可以进行改进和优化，以部署在临床研究环境中。
