Ezan E, Ducancel F, Gillet D, Drevet P, Ménez A, Grognet J M, Boulain J C
Service de Pharmacologie et d'Immunologie, DRIPP Bât 136, Commissariat à l'Energie Atomique, CE/Saclay, Gif-sur-Yvette, France.
J Immunol Methods. 1994 Mar 10;169(2):205-11. doi: 10.1016/0022-1759(94)90264-x.
A competitive enzyme immunoassay for rat prolactin using an immunogen and a tracer obtained by recombinant DNA technology is described. Polyclonal antibodies were raised in rabbits immunized with a purified chimeric protein consisting of rat prolactin fused with two synthetic immunoglobulin G binding domains derived from staphylococcal Protein A. The enzymatic tracer was obtained using an expression system which permits insertion of rPRL DNA sequence in the alkaline phosphatase gene. Antibodies and tracer were used to develop a solid-phase competitive immunoassay for the measurement of rat prolactin in plasma with a minimal detectable concentration of 0.5 ng/ml. The mean intra- and inter-assay coefficients of variation were 7.8 and 13.2%, respectively. Rat plasma concentrations measured with this assay correlated well with those obtained with a conventional enzyme immunoassay (r = 0.993, slope = 1.037, n = 24).
本文描述了一种使用免疫原和通过重组DNA技术获得的示踪剂来检测大鼠催乳素的竞争性酶免疫测定法。用一种纯化的嵌合蛋白免疫兔子制备多克隆抗体，该嵌合蛋白由大鼠催乳素与两个源自葡萄球菌蛋白A的合成免疫球蛋白G结合结构域融合而成。酶示踪剂是使用一种表达系统获得的，该系统允许将rPRL DNA序列插入碱性磷酸酶基因中。抗体和示踪剂用于开发一种固相竞争性免疫测定法，用于测量血浆中的大鼠催乳素，最低可检测浓度为0.5 ng/ml。测定的平均批内和批间变异系数分别为7.8%和13.2%。用该测定法测得的大鼠血浆浓度与用传统酶免疫测定法测得的浓度相关性良好（r = 0.993，斜率 = 1.037，n = 24）。
