Mandal C, Ali N
Indian Institute of Chemical Biology, Calcutta.
Steroids. 1988 Nov-Dec;52(5-6):551-60. doi: 10.1016/0039-128x(88)90122-5.
The preparation of high affinity and high specificity polyclonal and monoclonal antibodies to estradiol is described. Monoclonal antibodies were derived from BALB/c mice hyperimmunized with estradiol-3-O-carboxymethyl ether conjugated to bovine serum albumin. Spleen cells were hybridized with mouse myeloma cells. Quite a few monoclonal antibodies showed very good affinity for estradiol. Extended immunization and hyperimmunization were essential for producing a greater number of positive clones secreting high affinity antibodies. Binding constants of the antisera and their cross-reactivities with related steroids were calculated. Both polyclonal and monoclonal antibodies showed very high affinity for estradiol exhibiting little or no cross-reactivities with structurally related steroids indicating that this site of linkage is a good choice for discriminating between differences at the 16-17 position in the D-ring. This monoclonal antibody (44.28.6), having negligible cross-reactivity with estriol and estrone, can be used for diagnostic purposes.
本文描述了针对雌二醇制备高亲和力和高特异性的多克隆抗体和单克隆抗体的方法。单克隆抗体源自用与牛血清白蛋白偶联的雌二醇 - 3 - O - 羧甲基醚超免疫的BALB/c小鼠。脾细胞与小鼠骨髓瘤细胞进行杂交。相当多的单克隆抗体对雌二醇显示出非常好的亲和力。延长免疫和超免疫对于产生更多分泌高亲和力抗体的阳性克隆至关重要。计算了抗血清的结合常数及其与相关类固醇的交叉反应性。多克隆抗体和单克隆抗体对雌二醇均显示出非常高的亲和力，与结构相关的类固醇几乎没有或没有交叉反应，这表明该连接位点是区分D环中16 - 17位差异的良好选择。这种与雌三醇和雌酮交叉反应可忽略不计的单克隆抗体（44.28.6）可用于诊断目的。
