Novel sandwich ELISA for human angiotensinogen
Novel sandwich ELISA for human angiotensinogen
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DOI：
10.1152/ajprenal.00090.2007
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发表时间：
2007-09-01
影响因子：
4.2
通讯作者：
Kobori, Hiroyuki
中科院分区：
文献类型：
--
作者：
Katsurada, Akemi;Hagiwara, Yoshiaki;Kobori, Hiroyuki
人血管紧张素原的新型夹心ELISA。美国肾脏生理学杂志293：F956-F960，2007年。首次发表于2007年6月6日; doi：10.1152/ajprenal.00090.2007。我们最近报道，尿血管紧张素原（U-AGT）排泄率提供了一个特定的指标，肾内的肾素-血管紧张素（ANG）系统（RAS）的状态，在ANG II依赖性高血压大鼠。当这被证明是适用于人类受试者，诊断测试，以确定那些高血压患者最有可能响应RAS阻断可以提供有用的信息，允许一个机制的理由选择一个优化的方法来治疗高血压患者。然而，目前还没有简单而准确的方法来测量人血管紧张素原（hAGT）。对于人类受试者的未来研究，我们开发了抗体和一个敏感和特异性的定量系统，使用夹心ELISA的hAGT。我们提出了两种抗hAGT的抗体：小鼠单克隆抗体和兔多克隆抗体。该ELISA的标准曲线呈高度线性（0.31-20 ng/ml）。相关系数> 0.99。健康志愿者的血浆血管紧张素原浓度范围为28至71 μ g/ml（n = 10）。UAGT与尿肌酐浓度的比值范围为5.0 - 30 μ g/g（n = 7）。批内和批间变异系数范围分别为4.4 - 5.5%和4.3 - 7.0%。该ELISA系统与尿蛋白样本中的主要蛋白质，如人血白蛋白、免疫球蛋白或转铁蛋白无交叉反应。此外，该系统与血管紧张素肽的交叉反应性也可以忽略不计。hAGT ELISA法可用于研究高血压患者UAGT与抗高血压药物反应性的关系。
Novel sandwich ELISA for human angiotensinogen. Am J Physiol Renal Physiol 293: F956-F960, 2007. First published June 6, 2007; doi:10.1152/ajprenal.00090.2007. -We recently reported that urinary excretion rates of angiotensinogen (U-AGT) provide a specific index of intrarenal renin-angiotensin (ANG) system (RAS) status in ANG II-dependent hypertensive rats. When this is shown to be applicable to human subjects, a diagnostic test to identify those hypertensive patients most likely to respond to an RAS blockade could provide useful information to allow a mechanistic rationale for selection of an optimized approach to treatment of hypertensive patients. However, simple and accurate methods to measure human angiotensinogen (hAGT) are unavailable. For future studies of human subjects, we developed antibodies and a sensitive and specific quantification system for hAGT using a sandwich ELISA. We raised two antibodies against hAGT: a mouse monoclonal antibody and a rabbit polyclonal antibody. The standard curve of this ELISA exhibited a high linearity ( 0.31-20 ng/ml). The correlation coefficient was > 0.99. Plasma angiotensinogen concentrations of healthy volunteers ranged from 28 to 71 mu g/ml ( n = 10). The ratio of UAGT to urinary creatinine concentration ranged from 5.0 to 30 mu g/g ( n = 7). Intra- and interassay coefficients of variation ranged from 4.4 to 5.5% and from 4.3 to 7.0%, respectively. This ELISA system had no cross-reactivity with major proteins in proteinuric urine samples, such as human albumin, immunoglobulin, or transferrin. Moreover, the cross-reactivity of the system with angiotensin peptides was also negligible. This hAGT ELISA will be a useful tool to investigate the relationship of UAGT and reactivity to antihypertensive drugs in hypertensive patients.