Quantitative measurement of a candidate serum biomarker peptide derived from α2-HS-glycoprotein, and a preliminary trial of multidimensional peptide analysis in females with pregnancy-induced hypertension.
Quantitative measurement of a candidate serum biomarker peptide derived from α2-HS-glycoprotein, and a preliminary trial of multidimensional peptide analysis in females with pregnancy-induced hypertension.
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源自α2-HS-糖蛋白的候选血清生物标志物肽的定量测量，以及妊娠高血压女性多维肽分析的初步试验。
DOI：
10.1177/0004563217717748
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发表时间：
2018
期刊：
影响因子：
2.2
通讯作者：
Araki Y.
中科院分区：
文献类型：
--
作者：
Hamamura K;Yanagida M;Ishikawa H;Banzai M;Yoshitake H;Nonaka D;Tanaka K;Sakuraba M;Miyakuni Y;Takamori K;Nojima M;Yoshida K;Fujiwara H;Takeda S;Araki Y.
目的建立定量酶联免疫吸附试验 PDA 039/044/071肽的ELISA系统，潜在的血清疾病生物标志物 妊娠高血压综合征（PIH）的DBMs，主要通过肽组识别 方法（BLOTCHIP®-质谱法（MS））。然而，我们的方法并没有扩展到 PDA 071（半胱氨酰α2-HS-糖蛋白341 -367），由于难以产生 针对肽的特异性抗体。本研究的目的是建立一个 使用液相色谱-多重反应的替代PDA 071定量系统 方法采用热/酸变性法纯化血清PDA 071， 开发了允许对其进行特异性定量的LC-MRM/MS方法。我们测量 血清PDA 071浓度，这些结果得到验证，包括通过 针对PDA 039（激肽原-1439-456）/044的三维（3D）绘图 结果采用热变性法成功地从血清中提取出PDA 071。最佳 开发了通过LC-MRM/MS进行定量的条件;测定的血清PDA 071 与BLOTCHIP®测定值良好相关。虽然PDA 071本身并没有 患者和对照组之间存在显著差异，PDA 039/044/071的3D绘图 肽浓度和Jackknife分类矩阵的构建 结论PDA 071和PDA 039/044联合分析可用于妊高征的诊断 诊断准确率，我们的方法将是有价值的，在未来 妊娠期高血压疾病的病理生理学研究。
PurposeWe previously attempted to develop quantitative enzyme-linked immunosorbent assay (ELISA) systems for the PDA039/044/071 peptides, potential serum disease biomarkers (DBMs) of pregnancy-induced hypertension (PIH), primarily identified by a peptidomic approach (BLOTCHIP®-mass spectrometry (MS)). However, our methodology did not extend to PDA071 (cysteinyl α2-HS-glycoprotein341–367), due to difficulty to produce a specific antibody against the peptide. The aim of the present study was to establish an alternative PDA071 quantitation system using liquid chromatography-multiple reaction monitoring (LC-MRM)/MS, to explore the potential utility of PDA071 as a DBM for PIH.MethodsWe tested heat/acid denaturation methods in efforts to purify serum PDA071 and developed an LC-MRM/MS method allowing for specific quantitation thereof. We measured serum PDA071 concentrations, and these results were validated including by three-dimensional (3D) plotting against PDA039 (kininogen-1439–456)/044 (kininogen-1438–456) concentrations, followed by discriminant analysis.ResultsPDA071 was successfully extracted from serum using a heat denaturation method. Optimum conditions for quantitation via LC-MRM/MS were developed; the assayed serum PDA071 correlated well with the BLOTCHIP® assay values. Although the PDA071 alone did not significantly differ between patients and controls, 3D plotting of PDA039/044/071 peptide concentrations and construction of a Jackknife classification matrix were satisfactory in terms of PIH diagnostic precision.ConclusionsCombination analysis using both PDA071 and PDA039/044 concentrations allowed PIH diagnostic accuracy to be attained, and our method will be valuable in future pathophysiological studies of hypertensive disorders of pregnancy.