雌二醇多克隆抗体的制备与鉴定  

Preparation and qualification of estradiol antigen and antiserum

作　　者：高富红[1] 戴小华[1] 吴国娟[1] 

机构地区：[1]北京农学院动物科学系,北京102206

出　　处：《畜牧与兽医》2007年第10期19-22,共4页Animal Husbandry & Veterinary Medicine

基　　金：北京市农委科技攻关资助项目"畜产品中药物残留快速检测技术及产品研究与开发"(20020103)

摘　　要：制备雌二醇完全抗原,并通过免疫家兔得到多克隆抗体,为下一步制备雌二醇单克隆抗体和雌二醇检测ELISA试剂盒奠定基础。试验以牛血清白蛋白(BSA)、卵清蛋白(OVA)为载体,采用碳化二亚胺法,合成了雌二醇(E2)的2种免疫偶合物:免疫原E2-BSA和包被原E2-OVA;通过紫外光谱定性证明偶合物偶联成功与否,并对偶合物的蛋白含量、结合比进行测定并以免疫原E2-BSA免疫家兔,制备多克隆抗体,用包被原E2-OVA进行ELISA,对多克隆抗体特异性及效价进行检测。结果表明成功合成了雌二醇人工抗原即免疫原E2-BSA和包被原E2-OVA,二者的蛋白浓度分别为5.455和7.533mg/mL,结合比分别为7:1和8:1;制备的多克隆抗体特异性好,血清效价为1:106。In order to develop enzyme immunoassay for estradiol, the antigen and antiserum of estradiol were prepared. Estradiol was coupled to bovine serum albumin （BSA） and ovalbumin （OVA） by using the ester activation method. The conjugates were identified by UV absorption method to calculate the coupling ratio and the protein concentration. E2-BSA was used as immunogen for injection of rabbits to prepare antiserum. The affinity and titer of antiserum were detected by using the E2 -OVA as coating antigen. The results indicated that estradiol antigen was synthesized successfully. The protein concentration of E2 to BSA was 5.455 mg,/mL and to OVA was 7.533 mg,/mL. The coupling ratio of E2 to BSA was 7：1 and to OVA was 8：1. The titer of antiserum was 1：10^6.

关 键 词：雌二醇 人工抗原 多克隆抗体 

分 类 号：S852.4[农业科学—基础兽医学]