Jüppner H, Brabant G, Kapteina U, Kirschner M, Klein H, Hesch R D
Biochem Biophys Res Commun. 1986 Sep 30;139(3):1215-23. doi: 10.1016/s0006-291x(86)80307-2.
A direct radioimmunoassay for the rapid and accurate detection of human ANP from unextracted plasma is described. The sensitivity was approximately 50 pg/ml, respectively 2.5 pg/tube, the intra-assay variation 4%, and the inter-assay variation less than 12%. Rat ANP (1-28, 5-25, 5-27 and 5-28), oxydized and reduced hANP as well as plasma samples from various patients run in parallel to the 1-28 hANP standard curve. These findings imply, that the antibody primarily recognizes the mid-region (amino acids 6-25) of the intact ANP, that the C-terminal portion further increases the immunoreactivity, and that circulating plasma hANP is reliably measured. Plasma hANP ranged from 50-166 pg/ml (mean +/- SD: 98.3 +/- 44.6) in healthy individuals, there was no significant difference between samples were drawn in upright or lying position, the apparent half-life of injected hANP was 5.65 minutes. Patients with liver cirrhosis revealed significantly higher hANP levels of 244.5 +/- 173.5 pg/ml. Patients with various forms of cardiac disease had hANP concentrations ranging from 50 to 1744 pg/ml, depending at least partially on the right atrial pressure. No difference was observed if the samples were drawn from either right or left intracardial locations. Our findings with this system demonstrate that hANP is reliably measured even without prior extraction.
本文描述了一种直接放射免疫分析法,用于从未经提取的血浆中快速准确地检测人ANP。灵敏度分别约为50 pg/ml(即2.5 pg/管),批内变异为4%,批间变异小于12%。将大鼠ANP(1-28、5-25、5-27和5-28)、氧化型和还原型人ANP以及来自不同患者的血浆样本与1-28人ANP标准曲线平行检测。这些结果表明,该抗体主要识别完整ANP的中间区域(氨基酸6-25),C末端部分进一步增强免疫反应性,并且循环血浆中的人ANP能够得到可靠检测。健康个体的血浆人ANP范围为50-166 pg/ml(均值±标准差:98.3±44.6),直立位或卧位采血的样本之间无显著差异,注射的人ANP的表观半衰期为5.65分钟。肝硬化患者的人ANP水平显著更高,为244.5±173.5 pg/ml。患有各种心脏病的患者,其人ANP浓度范围为50至1744 pg/ml,至少部分取决于右心房压力。从右心或左心腔内采集样本未观察到差异。我们使用该系统的研究结果表明,即使不进行预先提取,人ANP也能得到可靠检测。