Viossat I, Chapelat M, Chabrier P E, Braquet P
Institut Henri-Beaufour, Les Ulis, France.
Arch Mal Coeur Vaiss. 1990 Jul;83(8):1263-5.
We have established a sensitive and specific radioimmunoassay for measurement of ET-1 in tissues and human plasma. Half maximal inhibition of binding of ET-1125 I was obtained at 6.6 pg/tube and the lowest concentration detectable was 0.785 pg/tube (p less than 0.001). Inter and intra-assay precisions were 14 and 9.6%, respectively. The antibody cross-reacted with ET-2 at 36.5%, ET-3 at 23.4% and Big ET at 9.9% but did not recognize non related vasoactive peptides. Immunoreactive ET-1 was extracted from human and rat tissues (lung, kidney) or human plasma using Sepak C18 columns. Human lung contains 2.3 ng/g ir-ET, corresponding mainly to synthetic ET-1 as revealed by reverse phase HPLC coupled to RIA. Rat lung and kidney ir-ET concentrations were 2.4 ng/g and 0.45 ng/g, respectively. Preliminary results indicated that the basal level of ir-ET in human plasma was 1.24 pg/ml. The RIA could be very useful to study the role of ET in various physiopathological states.
我们已经建立了一种灵敏且特异的放射免疫分析法,用于测量组织和人体血浆中的内皮素-1(ET-1)。ET-1 125I结合的半数最大抑制浓度为6.6 pg/管,可检测到的最低浓度为0.785 pg/管(p<0.001)。批间和批内精密度分别为14%和9.6%。该抗体与ET-2的交叉反应率为36.5%,与ET-3的交叉反应率为23.4%,与大内皮素的交叉反应率为9.9%,但不识别无关的血管活性肽。使用Sepak C18柱从人体和大鼠组织(肺、肾)或人体血浆中提取免疫反应性ET-1。人肺中含有2.3 ng/g免疫反应性ET,主要对应于反相高效液相色谱结合放射免疫分析法所显示的合成ET-1。大鼠肺和肾的免疫反应性ET浓度分别为2.4 ng/g和0.45 ng/g。初步结果表明,人体血浆中免疫反应性ET的基础水平为1.24 pg/ml。这种放射免疫分析法对于研究内皮素在各种生理病理状态中的作用可能非常有用。