| 莱克多巴胺单克隆抗体的研制及阻断ELISA检测方法的建立 |
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| 引用本文： | 张海棠,王自良,邓瑞广,钟华,范国英.莱克多巴胺单克隆抗体的研制及阻断ELISA检测方法的建立[J].核农学报,2008,22(6):904-908. |
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| 作者姓名： | 张海棠 王自良 邓瑞广 钟华 范国英 |
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| 作者单位： | [1]河南科技学院动物科学学院,河南新乡453003 [2]河南省动物免疫学重点实验室,河南郑州450002 |
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| 基金项目： | “十一五”国家科技支撑计划重大项目”农药、兽药抗体库建立”资助(2006BAK02A21/1) |
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| 摘 要： | 用混合酸酐法将莱克多巴胺(RAC)偶联于牛血清白蛋白(BSA),合成免疫原BSA-RAC,并用紫外和凝胶电泳鉴定;用BSA-RAC免疫Balb/C小鼠,细胞融合技术建立抗RAC单克隆抗体(mAb)杂交瘤细胞株,体内诱生腹水法制备RAC mAb。应用RAC mAb研制RAC残留快速检测阻断ELISA试剂盒(RAC-Kit),并测定其性能。结果表明,BSA-RAC偶联成功,分子结合比为1∶24.5;筛选出3株杂交瘤细胞,其中最好的4D8株的亲和常数Ka为1.65×1010L/mol。RAC-Kit的标准曲线呈典型的S型,符合4参数logit曲线拟合,线性范围为1~170μg/L,灵敏度为0.52μg/L,检测限为1μg/L,饲料样、猪尿样的平均添加回收率分别为91.1%和89.2%,平均批内和批间变异系数<15%,与多巴酚丁胺的交叉反应率为22.3%,与其他化合物无交叉反应,RAC-Kit在4℃可保存180d。
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| 关 键 词： | 莱克多巴胺 人工免疫原 单克隆抗体 阻断ELISA 快速检测试剂盒 |
| 收稿时间： | 2009-12-31 |
| 修稿时间： | 2009-12-31 |
GENERATION OF MONOCLONAL ANTIBODY AND DEVELOPMENT OF BLOCKING ELISA KIT FOR RACTOPAMINE |
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| ZHANG Hai-Tang,WANG Zi-liang,DENG Riu-Guang,ZHONG Hua,FAN Guo-Ying.GENERATION OF MONOCLONAL ANTIBODY AND DEVELOPMENT OF BLOCKING ELISA KIT FOR RACTOPAMINE[J].Acta Agriculturae Nucleatae Sinica,2008,22(6):904-908. |
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| Authors: | ZHANG Hai-Tang WANG Zi-liang DENG Riu-Guang ZHONG Hua FAN Guo-Ying |
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| Abstract: | Mixed anhydridewas used to conjugate ractopamine(RAC) to bovine serum albumin(BSA) and obtained artificial immunogen BSA-RAC identified by ultraviolet and SDS-PAGE.Balb/C mice were immunized with BSA-RAC and hybridoma lines that secrete monoclonal antibody against RAC(RAC mAb) were generated with cell fusion.A blocking ELISA kit for detection RAC(RAC-Kit)was developed with RAC mAb and its traits were tested.The results showed that BSA-RAC was synthesized successfully and its conjugation ratio of RAC to BSA ... |
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| Keywords: | ractopamine artificial immunogen monoclonal antibody blocking ELISA rapid test kit |
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