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2025年4月6日 星期日
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抗苯巴比妥单克隆抗体杂交瘤细胞株的筛选及ciELISA试剂盒的研制
王自良;张改平;张海棠;杨艳艳;王艳荣;柴书军
1.河南科技学院动物科学学院, 新乡 453003; 2.河南省动物免疫学重点实验室, 郑州 450002
Screening of Hybridoma Lines of Monoclonal Antibody and Development of ciELISA Kit for Rapid Detection of Phenobarbital
WANG Zi-liang;ZHANG Gai-ping;ZHANG Hai-tang;YANG Yan-yan;WANG Yan-rong; CHAI Shu-jun
1. College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China;
2. Henan Provincial Key Laboratory of Animal Immunology, Zhengzhou 450002, China
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摘要 用BSA-pAPB免疫Balb/c小鼠,用细胞融合技术制备并用间接ELISA和阻断ELISA筛选抗苯巴比妥单克隆抗体(PB mAb)杂交瘤细胞株,体内诱生腹水法生产PB mAb,应用PB mAb研制PB残留竞争ELISA(ciELISA)快速检测试剂盒(PB-Kit),并测定其性能。结果表明,筛选出3株杂交瘤细胞,最好的3F6-C4株的PB mAb间接ELISA效价为1∶6.4×105,亲和常数(Ka)为1.96×1010 L/moL,半数抑制浓度(IC50)为5.7 μg/L,与巴比妥的交叉反应率(CR%)12.4%,与其它化合物无CR;PB-Kit的线性检测范围1.0~81 μg/L,灵敏度0.75 μg/L,检测限1 μg/L;饲料样和猪尿样的平均添加回收率85.8%和91.3%,平均批内和批间变异系数均<15%。PB-Kit具有快速、敏感、特异、简便等特点,适合PB残留快速检测的推广应用。
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王自良
张改平
张海棠
杨艳艳
王艳荣
柴书军
关键词 苯巴比妥 杂交瘤细胞 单克隆抗体 ciELISA 快速检测试剂盒    
AbstractBalb/c mice were immunized with BSA-pAPB and hybridoma lines secreted monoclonal antibody against phenobarbital (PB mAb) were generated with cell fusion and filtered by indirect ELISA and blocking ELISA. A ciELISA kit for detection PB (PB-Kit) was developed with PB mAb and its traits were tested. The results showed that three hybridoma lines were filtered and the best one was 3F6-C4, which secreted PB mAb with indirect ELISA titers of 1∶6.4×105 in ascites. PB mAb had a high affinity constant(Ka) with 1.96×1010 L/mol, a good sensitivity with an IC50 of 5.7 μg/L to PB, 12.4% cross-reactivity to barbiturate sodium and little or no cross-reactivity to other compounds. The PB-Kit had the linear detection range of 1.0 to 81 μg/L, the sensitivity of 0.75 μg/L and the detection limit of 1.0 μg/L. The recoveries of PB spiked in feed and swine urine were 85.8% and 91.3% respectively. The precision and accuracy of the assay determined by inter-assay and intra-assay coefficient variation was both below 15%. The PB-Kit possesses rapidity, sensitivity, specificity and briefness. That is proved to be used for the rapid detection of PB residues in animal food.
Key wordsphenobarbital    hybridoma line    monoclonal antibody    ciELISA    rapid detection kit
收稿日期: 2005-11-21     
ZTFLH:  S184  
  S188  
通讯作者: 张改平   
引用本文:   
王自良;张改平;张海棠;杨艳艳;王艳荣;柴书军. 抗苯巴比妥单克隆抗体杂交瘤细胞株的筛选及ciELISA试剂盒的研制[J]. , 2006, 14(5): 711-715.
WANG Zi-liang;ZHANG Gai-ping;ZHANG Hai-tang;YANG Yan-yan;WANG Yan-rong; CHAI Shu-jun. Screening of Hybridoma Lines of Monoclonal Antibody and Development of ciELISA Kit for Rapid Detection of Phenobarbital. , 2006, 14(5): 711-715.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2006/V14/I5/711
 
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