| 吡虫啉残留酶联免疫吸附检测方法的建立 |
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| 引用本文： | 李广领,姜金庆,陈锡岭,张淑静,王自良. 吡虫啉残留酶联免疫吸附检测方法的建立[J]. 食品科学, 2011, 32(12): 228. DOI: 10.7506/spkx1002-6630-201112050 |
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| 作者姓名： | 李广领 姜金庆 陈锡岭 张淑静 王自良 |
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| 作者单位： | 河南科技学院资源与环境学院; |
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| 基金项目： | “十一五”国家科技支撑计划重大项目(2006BAK02A21/1); 河南省高校科技创新人才支持计划项目(2010HASTIT026) |
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| 摘 要： | 通过化学修饰合成吡虫啉(imidacloprid,IMI)人工半抗原,采用碳二亚胺法(EDC)将该抗原与牛血清蛋白和卵清蛋白偶联成功制备分子结合比合理的免疫抗原(IMI-BSA)和包被抗原(IMI-OVA)。对经IMI-BSA免疫的6周龄Balb/c实验鼠的免疫抗血清进行间接酶联免疫吸附和阻断酶联免疫吸附,初步探明抗吡虫啉多克隆抗体(IMI-pAb)的免疫学特性。在此基础上,采用聚乙二醇(PEG)介导细胞融合技术进行了免疫鼠脾细胞和骨髓瘤细胞融合,通过阳性杂交瘤细胞筛选和克隆化培养,获得3C8杂交瘤细胞株。结果显示:该3C8杂交瘤细胞株具有高效价、高亲和力和强特异性的特点;通过方阵滴定法确定最佳抗原包被浓度和最佳抗体稀释倍数,建立基于吡虫啉单克隆抗体(IMI-mAb)的IMI残留阻断酶联免疫吸附,其线性范围为7.48×10-6～3.24×10-4mg/mL(R2=0.9928),最低检测限为8.00×10-6mg/mL。
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| 关 键 词： | 吡虫啉 单克隆抗体 残留 酶联免疫吸附测定法 |
Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Imidacloprid Residues |
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| LI Guang-Ling,JIANG Jin-qing,CHEN Xi-ling,ZHANG Shu-jing,WANG Zi-liang. Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Imidacloprid Residues[J]. Food Science, 2011, 32(12): 228. DOI: 10.7506/spkx1002-6630-201112050 |
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| Authors: | LI Guang-Ling JIANG Jin-qing CHEN Xi-ling ZHANG Shu-jing WANG Zi-liang |
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| Affiliation: | LI Guang-Ling,JIANG Jin-qing,CHEN Xi-ling,ZHANG Shu-jing,WANG Zi-liang(School of Resources and Environmental Science,Henan Institute of Science and Technology,Xinxiang 453003,China) |
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| Abstract: | Chemical modification method was employed to synthesize imidacloprid(IMI) hapten.The IMI hapten was then coupled with BSA and OVA to prepare immunogen(IMI-BSA) and coating antigen(IMI-OVA) by EDC method.Optimal coupling ratios between the IMI hapten and the two carrier proteins were eventually achieved.In order to define immunological characteristics of IMI,anti-IMI serum produced by immunized six-week-old Balb/c mice was tested by indirect ELISA and blocking ELISA.Meanwhile,spleen cells of immunized mice w... |
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| Keywords: | imidacloprid monoclonal antibody residue enzyme linked immunosorbent assay(ELISA) |
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