Zia P, Zipser R, Speckart P, Horton R
J Lab Clin Med. 1978 Sep;92(3):415-22.
Antisera generated toward PGE was obtained from rabbits immunized with PGE2 conjugated to bovine thyroglobulin by the carbodiimide reaction. The specificity of the antibody is such that only PGE1 and PGE2 has significant cross-reactions. 13, 14-Dihydro and 15-keto PG's did not react. An RIA capable of measuring 6 pg of PGE2 was developed to measure PGE in human urine. Urine samples adjusted with buffer to pH-5 are extracted with 5% MeOH in CH2CI2 and chromatographed through Sephadex LH-20 columns. The second-antibody technique is used to separate bound from free. This urine method yields blank values of 2 +/- 2 pg per sample, with a between-assay precision determined by duplicate analysis of 14% and intra-assay precision of 8%. The mean urinary excretion rate is 500 +/- 74 (S.E.), +/- 209 (S.D.) ng/day (n = 8) in men and 300 +/- 70, +/- 242 (n = 12) in women. These values are in agreement with those reported by others using gas chromatography-mass spectrometry of receptor assay with hepatic plasma membranes. Patients in a high-renin state, whether normotensive or hypertensive, have elevated PGE in urine. These studies suggest a relationship between the renin-angiotensin system and renal PG's in man.
通过碳二亚胺反应,用与牛甲状腺球蛋白偶联的前列腺素E2免疫兔子,获得了针对前列腺素E的抗血清。该抗体的特异性在于只有前列腺素E1和前列腺素E2有显著的交叉反应。13,14 - 二氢和15 - 酮前列腺素不发生反应。开发了一种能够测量6皮克前列腺素E2的放射免疫分析法来测定人尿中的前列腺素E。用缓冲液将尿样调节至pH - 5,用5%甲醇的二氯甲烷溶液萃取,然后通过葡聚糖凝胶LH - 20柱进行色谱分析。采用第二抗体技术分离结合态和游离态。这种尿样检测方法每个样品的空白值为2±2皮克,通过重复分析确定的批间精密度为14%,批内精密度为8%。男性的平均尿排泄率为500±74(标准误),±209(标准差)纳克/天(n = 8),女性为300±70,±242(n = 12)。这些值与其他人使用气相色谱 - 质谱法或肝细胞膜受体分析法所报告的值一致。处于高肾素状态的患者,无论血压正常与否,尿中前列腺素E水平都会升高。这些研究表明,人类肾素 - 血管紧张素系统与肾脏前列腺素之间存在关联。