一种用于炔雌醇放射免疫测定的单克隆抗体的研制。

Development of a monoclonal antibody for use in radioimmunoassay of ethynylestradiol.

作者信息

Agasan A L, Briggs M H, Hewish D R, Stewart B

出版信息

Steroids. 1986 Apr-May;47(4-5):295-306. doi: 10.1016/0039-128x(86)90099-1.

Abstract

Production of monoclonal antibodies (MABs) to a 17 alpha-ethynyl-1,3,5 (10)-estratriene-3,17 beta-diol (ethynylestradiol, EE2) hapten is described. Culture antibodies generated by hybridized cells tested in enzyme-linked immunosorbent assay (ELISA) bound the 6-oxoethynyl-estradiol-6-(0-carboxymethyl) oxime-bovine serum albumin conjugate (EE2-6-0 CMO-BSA) but not BSA. On radioimmunoassay (RIA) with tritiated ethynylestradiol (3H-EE2), some of the antibodies demonstrated high binding affinity (Ka = 2.8 X 10(10) M-1) to EE2. Estradiol-17 beta, estrone and estriol did not show any displacement of 3H-EE2 from the MABs even at high concentration (1 X 10(4) ng/mL). Among the widely used progestins, norethynodrel and norethisterone exhibited considerable cross-reactivity (25.7-100% and 0.3-54.1%, respectively) but not levonor-gestrel with the MABs. The high affinity demonstrated by the MABs to EE2 3-sulfate but not to EE2 17-sulfate and EE2 3,17-disulfate suggests the dominant role of the 17 beta-hydroxyl group in immunogenicity.

摘要

本文描述了针对17α-乙炔基-1,3,5（10）-雌三烯-3,17β-二醇（乙炔雌二醇，EE2）半抗原的单克隆抗体（MABs）的制备。在酶联免疫吸附测定（ELISA）中测试杂交细胞产生的培养抗体与6-氧代乙炔基-雌二醇-6-（O-羧甲基）肟-牛血清白蛋白结合物（EE2-6-O CMO-BSA）结合，但不与牛血清白蛋白结合。在用氚标记的乙炔雌二醇（3H-EE2）进行放射免疫测定（RIA）时，一些抗体对EE2表现出高结合亲和力（Ka = 2.8×10（10）M-1）。即使在高浓度（1×10（4）ng/mL）下，17β-雌二醇、雌酮和雌三醇也未显示出从MABs中取代3H-EE2的情况。在广泛使用的孕激素中，炔诺酮和炔诺酮表现出相当大的交叉反应性（分别为25.7 - 100%和0.3 - 54.1%），但左炔诺孕酮与MABs没有交叉反应。MABs对EE2 3-硫酸盐表现出高亲和力，但对EE2 17-硫酸盐和EE2 3,17-二硫酸盐没有亲和力，这表明17β-羟基在免疫原性中起主导作用。