磺胺甲噁唑单克隆抗体的研制及免疫金标试纸检测方法的建立  

Developments of the Monoclonal Antibodies against Sulfamethoxazole and Immune Colloidal Gold-labeled Strip for Rapid Detection of Sulfamethoxazole

作　　者：刘宣兵[1,2] 张改平[1] 刘庆堂[1] 彩鸿翔[1,2] 杨继飞[1] 滕蔓[1] 邓瑞广[1] 侯玉泽[2] 王自良[3] 

机构地区：[1]河南省农业科学院动物免疫学重点实验室,河南郑州450002 [2]河南科技大学食品与生物工程学院,河南洛阳471003 [3]河南科技学院动物科学学院,河南新乡453003

出　　处：《农业生物技术学报》2009年第1期18-23,共6页Journal of Agricultural Biotechnology

基　　金：“十一五”国家科技支撑计划(No.2006BAK02A21)资助

摘　　要：用磺胺甲噁唑人工免疫原(BSA-SMZ)免疫BALB/C小鼠(Mus muscalus),细胞融合技术筛选抗磺胺甲噁唑单克隆抗体(SMZmAb)杂交瘤细胞,体内诱生腹水法制备SMZ mAb,并鉴定其免疫学特性;应用SMZmAb研制SMZ残留快速检测金标试纸(SMZ-strip),并测定其性能。结果表明,筛选出3株杂交瘤细胞,其中最好的3G6株间接ELISA效价细胞培养上清为1∶8.1×102,腹水为1∶6.4×105,亲和常数(Ka)为3.07×109 L/mol,对SMZ的半数抑制浓度(IC50)为12.4μg/L,与磺胺嘧啶的交叉反应率(CR%)为2.84%,与其它磺胺类药物无交叉反应;SMZ-strip目测检测限为6μg/L;其敏感性与竞争ELISA试剂盒相当,符合率为100%;SMZ-strip具有快速、敏感、特异、简便等特点,适合于SMZ残留快速检测的推广应用。Balb/C mice （Mus muscalus） were immunized with bovine serum albumin and sulfamethoxazole （BSA-SMZ）, hybridoma lines that secrete monoclonal antibody against sulfamethoxazole（SMZ mAb） were filtered with cell fusion and the immunological traits of SMZmAb were characterized. A strip for detection of SMZ （SMZ-strip） was developed with SMZmAb and its traits were tested. The results showed that three hybridoma lines were filtered and the best one was 3G6 which secreted SMZ mAb with indirect ELISA titers of 1：8.1×10^2 in supematant and 1：6.4×10^5 in ascites. SMZ mAb had a high affinity constant （Ka） with 3.07×10^9 L/mol, a good sensitivity with an IC50 of 12.4μg/L to SMZ, 2.84% cross-reactivity to sulfadiazine and little or no cress-reactivity to other compounds. Its detection limit was 6μg/L and its sensitivity was as same as that of competitive ELISA-kit and its coincidence rate was 100% compared with ELISA-kit. The SMZ-strip possesses rapidity, sensitivity, specificity and briefness and is proved to be used for the rapid test of SMZ residues.

关 键 词：磺胺甲噁唑 细胞融合 单克隆抗体 胶体金免疫层析试验 快速检测试纸 

分 类 号：S188[农业科学—农业基础科学]