| 吡虫啉的酶联免疫吸附分析方法研究 |
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| 引用本文： | 彭方毅,姜海蓉,陈远翔,陈胜珍,林治华,廖璞,何苗,施汉昌,蔡强. 吡虫啉的酶联免疫吸附分析方法研究[J]. 分析化学, 2010, 38(12). DOI: 10.3724/SP.J.1096.2010.01737 |
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| 作者姓名： | 彭方毅 姜海蓉 陈远翔 陈胜珍 林治华 廖璞 何苗 施汉昌 蔡强 |
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| 作者单位： | 重庆理工大学药学与生物工程学院,重庆,400050;重庆市临床检验中心,重庆,400014;清华大学环境模拟与污染控制国家重点联合实验室,北京,100084;浙江清华长三角研究院生态环境研究所,嘉兴,314001 |
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| 基金项目： | 国家863项目(No.2007AA062419); 重庆市科委自然基金(Nos.2008BB5331,2009BB7231); 重庆市教委自然基金(Nos.KJ090614,KJ100828); 清华大学环境模拟与污染控制国家重点联合实验室专项经费(No.09K05ESPCT)资助项目 |
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| 摘 要： | 通过碳二亚胺法将吡虫啉交联于牛血清蛋白(BSA)作为免疫抗原,混合酸酐法将吡虫啉交联于卵清蛋白(OVA)作为包被抗原,免疫Balb/c小鼠,采用B细胞杂交瘤技术,经免疫、融合、筛选、克隆,得到抗吡虫啉单克隆抗体,抗体亚类为IgG1,制备的单克隆抗体效价达1×107,确定了吡虫啉酶联免疫吸附分析方法(ELISA)的最佳工作条件,建立了定量测定吡虫啉的间接竞争ELISA方法。本方法的IC50为(15.12±1.28)μg/L,检出限为(1.76±0.02)μg/L。与其它吡虫啉结构类似物无交叉反应。批内相对标准偏差为4.5%;批间相对标准偏差5.1%,饮用水、重庆理工大学地下水和重庆市花溪河地表水平均添加回收率分别为102%,97%和85%。本研究建立了一种快速检测环境水中吡虫啉残留的方法。
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| 关 键 词： | 吡虫啉 半抗原 酶联免疫分析法 单克隆抗体 |
An Enzyme-linked Immunosorbent Assay for Detection of Imidacloprid |
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| PENG Fang-Yi,JIANG Hai-Rong,CHEN Yuan-Xiang,CHEN Sheng-Zhen,LIN Zhi-Hua,LIAO Pu,HE Miao,SHI Han-Chang,CAI Qiang. An Enzyme-linked Immunosorbent Assay for Detection of Imidacloprid[J]. Chinese Journal of Analytical Chemistry, 2010, 38(12). DOI: 10.3724/SP.J.1096.2010.01737 |
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| Authors: | PENG Fang-Yi JIANG Hai-Rong CHEN Yuan-Xiang CHEN Sheng-Zhen LIN Zhi-Hua LIAO Pu HE Miao SHI Han-Chang CAI Qiang |
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| Affiliation: | PENG Fang-Yi1,JIANG Hai-Rong 1,CHEN Yuan-Xiang1,CHEN Sheng-Zhen1,LIN Zhi-Hua1,LIAO Pu 2,HE Miao3,SHI Han-Chang3,CAI Qiang4 1(School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400050) 2(Clinical Laboratory Center of Chongqing,Chongqing 400014) 3(State Key Joint Laboratory of Environmental Simulation and Pollution Control,Tsinghua University,Beijing 100084) 4(Department of Environmental Technology and Ecology,Yangtze Delta Region Institute of Tsinghua University,Jiaxing 314000... |
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| Abstract: | To measure imidacloprid(IMI) in water,the hapten(imidacloprid) was conjugated to bovine serum albumin(BSA) with the carbodiimide method to form immunogen IMI-BSA;the hapten imidacloprid was conjugated to ovalbumin(OVA) with mixed carbonic anhydride method to form the coating antigens.Balb/c mice were immunized intraperitoneally with as prepare immunogen.Hybridoma cell lines secreting monoclonal antibodies(McAbs) against imidacloprid,which was obtained through routine hybridoma technology and ELISA screening... |
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| Keywords: | Imidacloprid Hapten Enzyme linked immunosorbent assay Monoclonal antibody |
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