Demonstrating the feasibility of large-scale development of standardized assays to quantify human proteins
Demonstrating the feasibility of large-scale development of standardized assays to quantify human proteins
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DOI：
10.1038/nmeth.2763
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发表时间：
2014-02-01
期刊：
影响因子：
48
通讯作者：
Paulovich, Amanda G.
中科院分区：
文献类型：
--
作者：
Kennedy, Jacob J.;Abbatiello, Susan E.;Paulovich, Amanda G.
多反应监测（MRM）质谱已成功应用于监测生物标本中的目标蛋白质，提高了分析可以配置为测量所有人类蛋白质的可能性。我们报告了一项试点研究的结果，该研究旨在测试大规模国际MRM检测的可行性。我们已经在三个实验室中配置和验证了645种新型MRM检测方法，这些方法代表了人类乳腺癌中表达的319种蛋白质。测定在>150种肽的组中多重化，并用于定量一组乳腺癌相关细胞系中的内源性分析物。中位测定精密度为5.4%，实验室间相关性高（R-2 > 0.96）。乳腺癌细胞系中的肽测量能够区分分子亚型并识别癌症蛋白质组中基因组驱动的变化。这些结果确立了大规模努力开发MRM测定资源的可行性。
Multiple reaction monitoring (MRM) mass spectrometry has been successfully applied to monitor targeted proteins in biological specimens, raising the possibility that assays could be configured to measure all human proteins. We report the results of a pilot study designed to test the feasibility of a large-scale, international effort for MRM assay generation. We have configured, validated across three laboratories and made publicly available as a resource to the community 645 novel MRM assays representing 319 proteins expressed in human breast cancer. Assays were multiplexed in groups of >150 peptides and deployed to quantify endogenous analytes in a panel of breast cancer-related cell lines. The median assay precision was 5.4%, with high interlaboratory correlation (R-2 > 0.96). Peptide measurements in breast cancer cell lines were able to discriminate among molecular subtypes and identify genome-driven changes in the cancer proteome. These results establish the feasibility of a large-scale effort to develop an MRM assay resource.