el-Gamal B A, Landon J, Abuknesha R A, Gallacher G, Perry L A
J Steroid Biochem. 1987 Mar;26(3):375-82. doi: 10.1016/0022-4731(87)90104-x.
Sheep were immunised with 11-deoxycortisol-21-hemisuccinate-bovine serum albumin (11-deoxycortisol-21-HS-BSA) or with 17-hydroxyprogesterone-7 alpha-carboxyethyl thioether-keyhole limpet haemocyanin (17-OHP-7 alpha-CETE-KLH) or with 17-OHP-3-(O-carboxymethyl)oxime-KLH (17-OHP-3-CMO-KLH). The resultant antisera were assessed using [3H]17-OHP and dextran-coated charcoal to separate the antibody bound and free fractions. All sheep produced antisera with an apparent affinity constant of from 1.4 to 6.6 X 10(9) 1/mol. Those raised against 11-deoxycortisol-21-HS-BSA had titres ranging from 1:12,000 to 1:78,000 but showed significant cross-reactivity with many of the steroids tested. Sheep immunised with 17-OHP-7 alpha-CETE-KLH had antisera titres of from 1:102,000 to 1:180,000 and only 17-hydroxypregnenolone cross-reacted significantly (10-20%). The best antisera were raised in sheep immunised with 17-OHP-3-CMO-KLH. Titres ranged from 1:168,000 to 1:390,000 and there were about 8 g/l of specific antibodies which cross-reacted 5.7% or less with 17-hydroxypregnenolone, and less than 0.5% with progesterone, 11-deoxycortisol and the other steroids studied. The antisera to 17-OHP-3-CMO-KLH were further assessed using [125I]17-OHP; titres ranged from 1:5,700,000 to 1:18,000,000 with affinity constants of from 1.67 to 2.5 X 10(10) 1/mol. They showed minimal or no cross-reactivity with the steroids studied. Reimmunisation after an 8-month interval produced antisera with a higher affinity constant and even lower cross-reactivity with other steroids.
用11 - 脱氧皮质醇 - 21 - 半琥珀酸 - 牛血清白蛋白（11 - 脱氧皮质醇 - 21 - HS - BSA）、17 - 羟孕酮 - 7α - 羧乙基硫醚 - 钥孔戚血蓝蛋白（17 - OHP - 7α - CETE - KLH）或17 - OHP - 3 - （O - 羧甲基）肟 - KLH（17 - OHP - 3 - CMO - KLH）对绵羊进行免疫。使用[³H]17 - OHP和葡聚糖包被的活性炭来分离抗体结合部分和游离部分，对所得抗血清进行评估。所有绵羊产生的抗血清的表观亲和常数为1.4至6.6×10⁹ 1/mol。那些针对11 - 脱氧皮质醇 - 21 - HS - BSA产生的抗血清效价范围为1:12,000至1:78,000，但与许多测试的类固醇显示出显著的交叉反应性。用17 - OHP - 7α - CETE - KLH免疫的绵羊的抗血清效价为1:102,000至1:180,000，只有17 - 羟孕烯醇酮有显著交叉反应（10 - 20%）。最好的抗血清是在用17 - OHP - 3 - CMO - KLH免疫的绵羊中产生的。效价范围为1:168,000至1:390,000，有大约8 g/l的特异性抗体，与17 - 羟孕烯醇酮的交叉反应率为5.7%或更低，与孕酮、11 - 脱氧皮质醇和其他研究的类固醇的交叉反应率小于0.5%。使用[¹²⁵I]17 - OHP对17 - OHP - 3 - CMO - KLH的抗血清进行进一步评估；效价范围为1:5,700,000至1:18,000,000，亲和常数为1.67至2.5×10¹⁰ 1/mol。它们与所研究的类固醇显示出最小或无交叉反应性。间隔8个月后再次免疫产生的抗血清具有更高的亲和常数，与其他类固醇的交叉反应性更低。
