Simple quantitation for potential serum disease biomarker peptides, primarily identified by a peptidomics approach in the serum with hypertensive disorders of pregnancy.
Simple quantitation for potential serum disease biomarker peptides, primarily identified by a peptidomics approach in the serum with hypertensive disorders of pregnancy.
复制标题
对潜在血清疾病生物标志物肽进行简单定量，主要通过肽组学方法在妊娠期高血压疾病的血清中进行鉴定。
DOI：
10.1177/0004563215583697
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发表时间：
2016
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;Nonaka D;Ishikawa H;Banzai M;Yanagida M;Nojima M;Yoshida K;Lee LJ;Tanaka K;Takamori K;Takeda S;Araki Y
背景资料：我们先前报道了使用新的肽组47分析方法的妊娠高血压综合征（PIH）的疾病生物标志物46（DBMs）的肽候选物，本研究的目的是建立一个48夹心酶联免疫吸附测定（ELISA）系统，用于定量49种此类肽，并验证其作为妊高征（包括妊娠期高血压/先兆子痫（GH/子痫））DBM的有用性。体育）。51方法：我们重点研究了激肽原-1438-456（PDA 039）、52激肽原-1439-456（PDA 044）和半胱氨酰α2-HS-糖蛋白341 -367（PDA 071）三个肽段。使用对每种肽特异的多克隆抗体（pAb），研究了夹心ELISA系统的合适条件。通过定量基质辅助激光解吸/电离飞行时间MS 56分析确认定量ELISA值为55。应用建立的ELISA方法对GH/PE患者血清和配对的57份健康孕妇血清进行了分析。结果：建立了PDA 039/044双抗体夹心ELISA定量检测的最佳条件。通过ELISA定量PDA 071失败，推测是由于60 pAb对天然肽的特异性的问题。Bland-Altman图显示ELISA法测定的血清PDA 039/044浓度与定量MS法测定的血清PDA 039/044浓度之间具有良好的相关性。虽然PDA 044水平在妊娠期间没有显示出显著变化，包括GH/PE患者，但患者血清PDA 039水平显著升高（P< 0.0001）。结论：首次建立了PDA 039的ELISA定量方法。PDA 039被证实其作为67例DBM的临床实用性
Background: We previously reported peptide candidates of disease biomarkers 46 (DBMs) for pregnancy-induced hypertension syndrome (PIH) using a novel peptidomic 47 analytical method, BLOTCHIP®-MS. The aim of this study was to establish a 48 sandwich enzyme-linked immunosorbent assays (ELISA) system for quantitation of 49 such peptides and to validate their usefulness as DBMs of PIH including gestational 50 hypertension/preeclampsia (GH/PE). 51Methods: We focused on three peptide fragments, kininogen-1438-456 (PDA039), 52 kininogen-1439-456 (PDA044) and cysteinyl α2-HS-glycoprotein341-367(PDA071). 53 Using polyclonal antibodies (pAbs) specific for each peptide, suitable conditions for the 54 sandwich ELISA system were investigated. The quantitative ELISA values were 55 confirmed by quantitative matrix assisted laser desorption/ionization time-of-flight MS 56 analyses. Using the established ELISA systems, sera from GH/PE patients and paired 57 serum samples from healthy pregnant females were analyzed. 58 Results: The optimum sandwich ELISA conditions for PDA039/044 quantitation were 59 developed. Quantitation of PDA071 by ELISA failed, presumably due to issues with 60 pAb specificity for the native peptide. Bland-Altman plots showed a satisfactory 61 correlation between the serum PDA039/044 concentration by ELISA and that by 62 quantitative MS analysis. Although the PDA044 level showed no significant change 63 during pregnancy, including in GH/PE patients, the serum PDA039 level was 64 significantly increased (P< 0.0001) in the patients. 65 Conclusions: The simple quantitation technology for PDA039 by ELISA was 66 established for the first time. PDA039 is confirmed its clinical utility as a DBM for 67