玉米赤霉醇单克隆抗体的制备及鉴定  

Preparation and identification of the monoclonal antibody to Zeranol

作　　者：王鹤佳[1] 曹振[2] 沈建忠[1] 史为民[1] 

机构地区：[1]中国农业大学动物医学院,北京海淀100094 [2]农业部兽医诊断中心,北京海淀100094

出　　处：《中国兽医杂志》2004年第10期3-5,共3页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金资助项目 ( 3 0 3 2 5 0 3 2 );"十五"国家科技攻关课题 ( 2 0 0 2 BA5 14 A-14

摘　　要：将玉米赤霉醇 (α- zearalanol,ZER)琥珀酰化得到 ZER- 7-半琥珀酸酯 ,再利用混合酸酐法与牛血清白蛋白 (BSA)偶联得到免疫原 ,以此免疫原免疫 BAL B/c小鼠 ,利用杂交瘤技术得到两株稳定分泌 ZER单克隆抗体的杂交瘤细胞株 1C1 2 和 9B4 ,经鉴定两株单抗亚型均为 Ig G1 ,间接 EL ISA测定腹水效价为 1∶ 3.2× 10 5。该抗体与同系物β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮、玉米赤霉烯酮的交叉反应率分别为 36 .2 5 % ,5 2 .73% ,0 .6 7% ,74 .36 % ,5 3.70 % ;与己烯雌酚、雌二醇、睾酮、克伦特罗的交叉反应率均小于 0 .1%。In this study, Zeranol(ZER) was coupled to carrier protein bovine serum albumin(BSA) using the mixed anhydride method. The ZER-BSA was used as immunogen. Two hybridomas producing anti-ZER monoclonal antibody were obtained by limited dilution, named 1C12 and 9B_4. The characterization of monoclonal antibody was performed by ELISA test. The results indicated: the subclass was IgG_1, the titer of ascitic fluid was 1∶3.2×10~5, the chromosomal numbers were 95～102. The cross-reactivities of McAb to zeranol,β-zearalanol,α-zearalenol, β-zearalenol, zearalanone, zearalenone were 36.25%,(52.73%),0.67%,74.36%, 53.70% respectively, and to diethylstilbestrol, 17β-estradiol, testosterone, clenbuterol were all below 0.1%.

关 键 词：玉米赤霉醇 单克隆抗体 制备 鉴定 杂交瘤技术 残留检测方法 

分 类 号：S816.7[农业科学—饲料科学] S859.84[农业科学—畜牧学]