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一种使用具有高特异性的抗体和用于前列腺素分离的葡聚糖凝胶LH - 20微柱的PGE2特异性放射免疫测定法。

A specific radioimmunoassay for PGE2 using an antibody with high specificity and a sephadex LH-20 microcolumn for the separation of prostaglandins.

作者信息

Christensen P, Leyssac P P

出版信息

Prostaglandins. 1976 Feb;11(2):399-420. doi: 10.1016/0090-6980(76)90161-1.

Abstract

Antiserum against PGE2 was raised in rabbits following immunization with prostaglandin-hen-gamma-globulin conjugate. The antiserum exhibited 14% cross reactivity with PGE1 and far less cross-reaction with heterologous prostaglandins. A microcolumn of Sephadex LH-20 was used for a partial, but sufficient separation of PGE2 from PGE1 and a complete separation from heterologous prostaglandins to ensure a specific RIA for PGE2. The precision of the method in the rage 10-500 picograms showed a coefficient of variation varying between 4 and 13%. The detection limit was 10 picograms corresponding to 15 pg/ml of PGE2 in serum. In order to demonstrate the validity of the method values obtained for non-diuretic rat renal venous serum were compared with those obtained using the isotope derivative method of Bojesen & Buckhave (1972) on the same samples. The concentrations of PGE2 obtained were 239 +/- 25 pg/ml and 250 +/- 58 pg/ml, respectively.

摘要

用前列腺素 - 鸡γ - 球蛋白偶联物免疫家兔后,制备了抗前列腺素E2(PGE2)抗血清。该抗血清与前列腺素E1(PGE1)有14%的交叉反应,与异源前列腺素的交叉反应则少得多。使用葡聚糖凝胶LH - 20微柱对PGE2与PGE1进行部分但足够的分离,并与异源前列腺素完全分离,以确保对PGE2进行特异性放射免疫分析(RIA)。该方法在10 - 500皮克范围内的精密度显示变异系数在4%至13%之间。检测限为10皮克,相当于血清中PGE2浓度为15 pg/ml。为证明该方法的有效性,将非利尿大鼠肾静脉血清所得值与使用Bojesen和Buckhave(1972年)的同位素衍生法对相同样本所得值进行比较。所得PGE2浓度分别为239±25 pg/ml和250±58 pg/ml。

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