Hindawi R K, Padfield P L
Prostaglandins Leukot Med. 1985 Apr;18(1):81-94. doi: 10.1016/0262-1746(85)90053-8.
A direct radioimmunoassay for urinary PGE2 has been developed, using a Pasteur Institute antibody and a gamma-labelled ligand. The assay does not require preliminary solvent extraction of the sample or any chromatographic steps. Gamma-labelled PGE2 of high specific activity was prepared by direct iodination of a PGE2-histamine derivative. Results obtained by the direct assay after serial dilution of urine samples were parallel to the standard curve. Accuracy of the method was determined by the recovery of added amounts of PGE2 to diluted urine (r = 0.094, y = 1.039x-0.535, n = 20). The lowest concentration of PGE2 distinguishable from zero was 0.063 pg/ml. The assay curve covered the range of 0.063-500 pg/ml. The assay was desensitized by increasing the concentration of both label and antiserum to obtain a curve covering the range of 2-1250 pg/ml. Intra-assay and interassay coefficient of variation were 8.6% and 12.5% respectively. Values obtained by the direct method were lower than those obtained by gas chromatography mass spectrometry (r = 0.43, y = 1.57x-3.52, n = 12) and by radioimmunoassay using 3H-PGE2 (r = 0.76, y = 0.93x+2.87, n = 12). Antibodies to PGE2 were produced in rabbits immunised with a conjugate of PGE2 covalently linked to thyroglobulin by carbodiimide reaction. The method was used to study the effect of urine flow and sodium intake on PGE2 excretion levels in normal volunteers. PGE2 excretion was flow dependent but did not vary with sodium excretion.
已开发出一种用于检测尿中前列腺素E2（PGE2）的直接放射免疫分析法，该方法使用巴斯德研究所的抗体和γ标记配体。该检测方法不需要对样品进行初步溶剂萃取或任何色谱步骤。通过对PGE2-组胺衍生物进行直接碘化制备了高比活度的γ标记PGE2。对尿样进行系列稀释后，直接检测法得到的结果与标准曲线平行。通过向稀释尿液中添加PGE2后的回收率来确定该方法的准确性（r = 0.094，y = 1.039x - 0.535，n = 20）。可与零区分的PGE2最低浓度为0.063 pg/ml。检测曲线覆盖0.063 - 500 pg/ml的范围。通过增加标记物和抗血清的浓度使检测方法脱敏，以获得覆盖2 - 1250 pg/ml范围的曲线。批内和批间变异系数分别为8.6%和12.5%。直接法获得的值低于气相色谱-质谱法（r = 0.43，y = 1.57x - 3.52，n = 12）和使用3H-PGE2的放射免疫分析法（r = 0.76，y = 0.93x + 2.87，n = 12）得到的值。用通过碳二亚胺反应将PGE2与甲状腺球蛋白共价连接而成的结合物免疫兔子，产生了抗PGE2抗体。该方法用于研究尿流和钠摄入量对正常志愿者PGE2排泄水平的影响。PGE2排泄量依赖于尿流，但不随钠排泄量而变化。
