| 脱氧雪腐镰刀菌烯醇免疫分析核心试剂特异性单克隆抗体的研制与特性研究 |
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| 引用本文： | 王海彬,李培武,张奇,张文,丁小霞,张兆威.脱氧雪腐镰刀菌烯醇免疫分析核心试剂特异性单克隆抗体的研制与特性研究[J].化学试剂,2012,34(9):771-776. |
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| 作者姓名： | 王海彬 李培武 张奇 张文 丁小霞 张兆威 |
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| 作者单位： | 1. 中国农业科学院油料作物研究所,湖北武汉,430062
2. 中国农业科学院油料作物研究所,湖北武汉430062;农业部生物毒素检测重点实验室,湖北武汉430062;农业部油料作物生物学与遗传育种重点实验室,湖北武汉430062;农业部油料及制品质量监督检验测试中心,湖北武汉430062;农业部油料产品质量安全风险评估实验室,湖北武汉430062 |
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| 基金项目： | 农业部“948”项目，国家自然科学基金项目，国家农业行业科技专项项目，国家科技支撑计划项目，中国农业科学院油料作物研究所所长基金项目，现代农业产业技术体系建设专项资金 |
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| 摘 要： | 采用1,1’-羰基二咪唑(CDI)活化法成功将脱氧雪腐镰刀菌烯醇分子上C-3和C-15位羟基与载体蛋白赖氨酸的氨基共价偶联得到免疫原DON-BSA和包被原DON-OVA,并用三硝基苯磺酸法对合成结果进行鉴定。以人工抗原免疫Balb/c小鼠,取小鼠脾细胞与SP2/O鼠骨髓瘤细胞融合,经筛选和多次亚克隆,得到了1株能稳定分泌脱氧雪腐镰刀菌烯醇抗体的单克隆细胞株1D5,并制备单克隆抗体腹水。经检测该抗体亚型为IgG1,亲和力常数Ka为8.33×107L/mol,交叉反应的试验结果显示该抗体与其他真菌毒素无交叉反应率,稳定性良好,为粮油食品中脱氧雪腐镰刀菌烯醇免疫快速检测技术建立及产品研发提供了关键试剂材料。
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| 关 键 词： | 呕吐毒素 人工抗原 单克隆抗体 |
Production and characterization of monoclonal antibody against deoxynivalenol |
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| WANG Hai-bin
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LI Pei-wu
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ZHANG Qi
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ZHANG Wen
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DING Xiao-xia
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ZHANG Zhao-wei.Production and characterization of monoclonal antibody against deoxynivalenol[J].Chemical Reagents,2012,34(9):771-776. |
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| Authors: | WANG Hai-bin LI Pei-wu ZHANG Qi ZHANG Wen DING Xiao-xia ZHANG Zhao-wei |
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| Affiliation: | 1,2,3,4,5(1.Oil Crops Research Institute,Chinese Academy of Agricultural Sciences,Wuhan 430062,China;2.Key Laboratory of Detection for Mycotoxins,Ministry of Agriculture,Wuhan 430062,China;3.Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture,Wuhan 430062,China;4.Quality Inspection and Test Center for Oilseeds Products MOA PRC,Wuhan 430062,China;5.Laboratory of Risk Assessment for Oilseeds Products MOA PRC,Wuhan 430062,China) |
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| Abstract: | Deoxynivalenol(DON) was activated by 1,1-dicyclohexylcarbodiimide(CDI),and conjugated to bovine serum albumin(BSA) and ovalbumin(OVA) as immunogen(DON-BSA) and coating antigen(DON-OVA).After fusing mouse myeloma cells(SP 2/0) with spleen cells from Balb/c mice which were immunized by the artificial antigen conjugated with BSA,screening and subclonal procedure,one hybridoma cell line 1D5 stably secreting monoclonal antibody(McAb) against DON was obtained.Isotype of the monoclonal antibody 1D5 was shown as IgG1.The results showed that the affinity constant was 8.33×107 L/mol,and no cross-reactivity was seen to the other tested mycotoxins(Fumonisin B1,Ochratoxin A,T-2 toxin,Zearalenone).The antibody also showed excellent stability,which could provide a key reagent material for development deoxynivalenol-specific immuno-assay technologies and products. |
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| Keywords: | deoxynivalenol artifical antigen monoclonal antibody |
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