抗黄曲霉毒素M1单克隆抗体的制备及定量ELISA方法的建立  

Development of monoclonal antibody against Aflatoxin M1 and the establishment of quantitative ELISA

作　　者：张园园[1] 裴世春[1] 才琳[1] 何娜[1] 赵识非[1] 霍贵成[2] 

机构地区：[1]黑龙江八一农垦大学食品学院,黑龙江大庆163319 [2]东北农业大学,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2008年第10期800-804,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省留学归国人员基金(Lc05c11)

摘　　要：采用黄曲霉毒素M1与牛血清白蛋白偶联物免疫8周龄BALB/c小鼠,4次免疫后,使小鼠产生免疫应答,取其脾细胞与Sp2/0骨髓瘤细胞融合、筛选,最终获得1株能稳定分泌抗黄曲霉毒素M1的杂交瘤细胞株系。其染色体平均计数为90±10对,间接ELISA检测该株系培养上清的效价为1∶6400,纯化腹水效价为5000×27,与黄曲霉毒素M1及其结构类似物黄曲霉毒素B1和脱氧雪腐镰刀菌烯醇的反应率分别为100%、7%、0%。经传30代后,抗体效价稳定。并在此基础上建立了间接竞争ELISA检测方法,该方法的最低检出浓度是0.08ng/mL,校正曲线的线性范围为0.08ng/mL～5ng/mL。该方法的加标回收率为80.0%～128.3%。Eight week BALB/c mice were immunized by Aflatoxin Ml-bovine serum albumin BSA conjugate （AFM1-BSA） and the spleen cells were fused with moue Sp2/0 myeloma cells. Positive hybridoma was detected by indirect competitive ELISA. One hybridoma cell line stably secreting monoclonal antibody against AFM1 was obtained. The McAb had a ascites titer of 5 000×27, and was highly specific against aflatoxin M1 （AFM1）. The minimum detection concentration of Aflatoxin M1 was 0.08 ng/mL with a linear range of 0.08 ng/mL to 5 ng/mL. The recovery of Afatoxin M1 was between 80.0 % to 128.3 %.

关 键 词：黄曲霉毒素M1 酶联免疫吸附测定 单克隆抗体 

分 类 号：R446.61[医药卫生—诊断学]