Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma.
Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma.
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DOI：
10.1038/nbt.1546
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发表时间：
2009-07
影响因子：
46.9
通讯作者：
Carr, Steven A.
中科院分区：
文献类型：
--
作者：
Addona, Terri A.;Abbatiello, Susan E.;Schilling, Birgit;Skates, Steven J.;Mani, D. R.;Bunk, David M.;Spiegelman, Clifford H.;Zimmerman, Lisa J.;Ham, Amy-Joan L.;Keshishian, Hasmik;Hall, Steven C.;Allen, Simon;Blackman, Ronald K.;Borchers, Christoph H.;Buck, Charles;Cardasis, Helene L.;Cusack, Michael P.;Dodder, Nathan G.;Gibson, Bradford W.;Held, Jason M.;Hiltke, Tara;Jackson, Angela;Johansen, Eric B.;Kinsinger, Christopher R.;Li, Jing;Mesri, Mehdi;Neubert, Thomas A.;Niles, Richard K.;Pulsipher, Trenton C.;Ransohoff, David;Rodriguez, Henry;Rudnick, Paul A.;Smith, Derek;Tabb, David L.;Tegeler, Tony J.;Variyath, Asokan M.;Vega-Montoto, Lorenzo J.;Wahlander, Asa;Waldemarson, Sofia;Wang, Mu;Whiteaker, Jeffrey R.;Zhao, Lei;Anderson, N. Leigh;Fisher, Susan J.;Liebler, Daniel C.;Paulovich, Amanda G.;Regnier, Fred E.;Tempst, Paul;Carr, Steven A.
候选生物标志物的验证依赖于针对靶蛋白的选择性检测而优化的特异性定量测定，并且越来越多地被视为发现管道中的关键步骤，其将无偏见的生物标志物发现与临床前验证联系起来。尽管个别实验室已证明多反应监测（MRM）结合同位素稀释质谱法可以定量血浆中的候选蛋白质生物标志物，但尚未证明这些试验在实验室之间的重现性和可转移性。我们描述了一个多实验室研究，以评估重现性，回收率，线性动态范围和检测限和定量的多重，基于MRM的测定，进行NCI-CPTAC。使用常见的材料和标准化的方案，我们证明了这些测定在实验室和仪器平台内和跨实验室和仪器平台具有高度的重现性，并且对未分级血浆中的低µg/ml蛋白质浓度敏感。我们提供数据和基准，使各个实验室可以比较其性能，并评估血浆生物标志物验证的新技术。
Verification of candidate biomarkers relies upon specific, quantitative assays optimized for selective detection of target proteins, and is increasingly viewed as a critical step in the discovery pipeline that bridges unbiased biomarker discovery to preclinical validation. Although individual laboratories have demonstrated that multiple reaction monitoring (MRM) coupled with isotope dilution mass spectrometry can quantify candidate protein biomarkers in plasma, reproducibility and transferability of these assays between laboratories have not been demonstrated. We describe a multilaboratory study to assess reproducibility, recovery, linear dynamic range and limits of detection and quantification of multiplexed, MRM-based assays, conducted by NCI-CPTAC. Using common materials and standardized protocols, we demonstrate that these assays can be highly reproducible within and across laboratories and instrument platforms, and are sensitive to low µg/ml protein concentrations in unfractionated plasma. We provide data and benchmarks against which individual laboratories can compare their performance and evaluate new technologies for biomarker verification in plasma.
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