吖啶酯标记的化学发光免疫分析优化参数评价

EVALUATION OF OPTIMAL PARAMETERS FOR ACRIDINIUM ESTER LABELED CHEMILUMINESCENE IMMUNOASSAY

以睾酮－３－羧甲氧基肟－牛血清白蛋白为免疫原，免疫家兔产生抗血清，１０－甲基吖啶－９－羧酸［４－（Ｎ－睾酮－３－羧甲氧基肟）－氨乙基］苯酯为示踪化合物，采用平衡法建立睾酮化学发光免疫分析，数据用四参数Ｌｏｇｉｓｔｉｃ函数处理．方法学考核标准曲线稳定，Ｒ为０．９９６７；ＥＤ５０为８２．１５ｎｇ／Ｌ；批内和批间ＣＶ分别为５．７８％和３．３％；平均回收率为１０７．２８％；可测范围为１９．３～２６４．６ｎｇ／Ｌ，与雌酮、雌二醇、雌三醇和孕酮的交叉反应率均小于１．３５％；非特异结合率为９．６４％．“桥效应”分析表明同位点、“桥”

Immunogen was testosterone 3 carboxymethyl oxime bovine serum albumin(To 3 CMO BSA), and tracer ligand was testosterone 3 carboxymethyl oxime 4 (10 methy) acridinium 9 carboxylate(To 3 CMO AEPMAC). A chemiluminescene immunoassay of testosterone(To CLIA) was established, on the equilibrium method, by raising the antiserum in rabbits against To 3 CMO BSA. The data was processed by the four parameters logistic function program. The intra and inter assay CV of To CLIA were 5 78% and 3 30%, respectively, the recovery rate, 107 28%. The antiserum provided a linear response from 19 3 to 264 6 ng/L with ED 50 of 82 15 ng/L and correlation coefficient of curve fitting, 0 9967. The cross reactivity with estrone, estradiol, estriol and progesterone were less 1 35%. Non specific bound rate was 9 64%. The highest affinity constant could be obtained by “bridge effect” analysis with same structure of the “bridge” in the same position of testosterone.