Absolute quantification of C-reactive protein in human plasma derived from patients with epithelial ovarian cancer utilizing protein cleavage isotope dilution mass spectrometry.
Absolute quantification of C-reactive protein in human plasma derived from patients with epithelial ovarian cancer utilizing protein cleavage isotope dilution mass spectrometry.
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DOI：
10.1021/pr800922p
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发表时间：
2009-02
影响因子：
4.4
通讯作者：
Muddiman, David C.
中科院分区：
文献类型：
--
作者：
Williams, D. Keith, Jr.;Muddiman, David C.
建立了蛋白裂解同位素稀释质谱法（PC-IDMS）测定人血浆中C反应蛋白（CRP）的方法。该方法是在不使用免疫亲和色谱法或尺寸排阻色谱法的情况下完成的，因为先前用于定量CRP的质谱方法已经使用。使用PC-IDMS通过1-D nano LC-MS/MS，使用30分钟梯度，使用三重四极杆质谱仪（以选择性反应监测（SRM）模式运行），分析了总计110份人血浆样本。将这种新开发方法的结果与由独立CLIA认证实验室进行的酶联免疫吸附试验（ELISA）产生的结果进行比较。这些结果的比较产生了R2 = 0.9708，这表明利用本文所述的方法成功地定量了来自人血浆的CRP。有趣的是，PC-IDMS方法提供的浓度值约为ELISA方法报告浓度的10倍，这表明在确定特定分析物的参考范围时，检测方法是一个重要考虑因素。此外，数据显示，随着上皮性卵巢癌（EOC）从I期进展到IV期，CRP的平均水平增加。
A method employing protein cleavage isotope dilution mass spectrometry (PC-IDMS) was developed for quantification of C-reactive protein (CRP) in human plasma. This method was completed without the use of immuno-affinity chromatography or size exclusion chromatography, as previous mass spectrometric methods for the quantification of CRP have employed. A total of 110 human plasma samples were analyzed with PC-IDMS via 1-D nano LC-MS/MS using a 30 minute gradient with a triple quadrupole mass spectrometer operated in selective reaction monitoring (SRM) mode. The results from this newly developed method were compared to results generated from an enzyme-linked immunosorbent assay (ELISA) performed by an independent CLIA certified laboratory. The comparison of these results generated a R2 = 0.9708 which indicates successful quantification of CRP from human plasma utilizing the methodology described herein. Interestingly, the PC-IDMS method provided concentration values that were ~10X the concentration reported by the ELISA method, which demonstrated that the method of detection is an important consideration when determining reference ranges of a particular analyte. In addition, data is shown that illustrates that as epithelial ovarian cancer (EOC) progresses from stage I to stage IV, mean levels of CRP increase.
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