The simultaneous quantification of candidate serum biomarker peptides for hypertensive disorders of pregnancy
The simultaneous quantification of candidate serum biomarker peptides for hypertensive disorders of pregnancy
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妊娠期高血压疾病候选血清生物标志物肽的同步定量
DOI：
10.1177/0004563219839084
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Araki Yoshihiko
中科院分区：
文献类型：
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作者：
Yanagida Mitsuaki;Hamamura Kensuke;Takamori Kenji;Araki Yoshihiko
背景妊娠期高血压疾病被定义为以妊娠 20 周后出现的高血压为特征的综合征。妊娠期高血压疾病的发病机制尚未完全阐明，最有效的治疗方法是终止妊娠。因此，应该开发早期预测诊断的方法来拯救母亲和孩子。方法之前，我们通过肽组学分析探索了妊娠期高血压疾病患者和正常孕妇之间浓度存在特异性差异的血清肽，并鉴定了七种候选标记肽。为了更可靠地量化标记肽，我们尝试使用液相色谱/串联质谱 (LC-MRM/MS) 通过多反应监测同时量化这些肽。合成了七种肽的非标记形式和稳定同位素标记形式作为标准品，并确定了其定量的多反应监测转变。结果由于反相柱对肽的保留取决于其疏水性，因此需要两种溶剂组合物来保留所有肽。在这些条件下，我们使用柱切换方法通过 LC-MRM/MS 检测肽。此外，我们使用稳定同位素稀释成功地定量了孕妇血清中的肽。结论我们的新肽组学方法对于肽，特别是那些没有特异性抗体作为检测工具的肽具有很大的价值。使用该系统，可以验证妊娠期高血压疾病患者的血清肽作为妊娠期高血压疾病的诊断标志物。此外，该方法有可能应用于其他血清肽的一般定量。
BackgroundHypertensive disorders of pregnancy are defined as syndromes characterized by high blood pressure that develop after 20 weeks of pregnancy. The pathogenesis of hypertensive disorders of pregnancy has not been fully elucidated, and the most effective treatment is the termination of pregnancy. Therefore, methods for an early predictive diagnosis should be developed to rescue both the mother and the child.MethodsPreviously, we explored the serum peptides whose concentration varied specifically between patients with hypertensive disorders of pregnancy and normal pregnant women using peptidomics analysis and identified seven candidate marker peptides. To quantify the marker peptides more reliably, we attempted to quantify these peptides simultaneously by multiple reaction monitoring using liquid chromatography/tandem mass spectrometry (LC-MRM/MS). Non-labelled and stable isotope-labelled forms of the seven peptides were synthesized as standards, and the multiple reaction monitoring transitions for their quantification were determined.ResultsAs the retention of the peptides by the reversed-phase column was dependent on their hydrophobicity, two solvent compositions were required for the retention of all peptides. Under these conditions, we detected the peptides by LC-MRM/MS using a column-switching method. Further, we succeeded in quantifying the peptides in the serum of pregnant women using stable isotope dilution.ConclusionOur new peptidomics method has great value for peptides, particularly those that have no specific antibody as a detection tool. Using this system, the serum peptides in patients with hypertensive disorders of pregnancy can be validated as diagnostic markers of hypertensive disorders of pregnancy. Further, this method can potentially be applied to the general quantification of other serum peptides.