Hennies M, Holtz W
Institute of Animal Husbandry and Genetics, Göttingen University, Germany.
J Immunol Methods. 1993 Jan 4;157(1-2):149-53. doi: 10.1016/0022-1759(93)90081-h.
The development of a competitive enzyme immunoassay for bovine growth hormone (bGH) is described. Antiserum, raised in rabbits and diluted 1/250,000, was preincubated with samples and free antibodies from the reaction mixture were immobilized using a microtiter plates coated with bGH. Bound antibodies remaining from the preincubation were visualized using a biotinylated second antibody as a bridge for subsequent amplification by an avidin-biotin-peroxidase complex. The measuring range was between 0.5 and 100 ng/ml. Cross-reactivity with other pituitary hormones was < 0.1%. The intra- and interassay coefficients of variations were 8.1 and 12.7%, respectively, and the recovery of added bGH was 110%. To validate the assay, two bull calves were treated with bGH-releasing factor. The response showed that there was an immediate rise in immunoreactive bGH which peaked after 5 and 15 min at 200 ng/ml and 60 ng/ml, respectively. This enzyme immunoassay is an economic and sensitive alternative to the established radioimmunoassay, and the first competitive enzyme immunoassay described for bGH.
本文描述了一种用于牛生长激素（bGH）的竞争性酶免疫测定法的开发。用兔制备的抗血清经1/250,000稀释后，先与样品预温育，然后使用包被有bGH的微量滴定板固定反应混合物中的游离抗体。预温育后剩余的结合抗体通过生物素化二抗作为桥梁，利用抗生物素蛋白-生物素-过氧化物酶复合物进行后续放大来显色。测量范围为0.5至100 ng/ml。与其他垂体激素的交叉反应率<0.1%。批内和批间变异系数分别为8.1%和12.7%，添加bGH的回收率为110%。为验证该测定法，对两头公牛犊用bGH释放因子进行处理。结果显示，免疫反应性bGH立即升高，分别在5分钟和15分钟时达到峰值，浓度分别为200 ng/ml和60 ng/ml。这种酶免疫测定法是已有的放射免疫测定法的一种经济且灵敏的替代方法，也是所描述的第一种用于bGH的竞争性酶免疫测定法。
