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当前位置: 首页 > 文章 > 新型抗植物病毒剂毒氟磷的酶联免疫分析方法 农药 2014,53 (11) 26-29
Position: Home > Articles > The Immunoassay Method for the Detection of Dufulin Agrochemicals 2014,53 (11) 26-29

新型抗植物病毒剂毒氟磷的酶联免疫分析方法

作  者：
卢平;李艳娇;丁晓燕
单  位：
贵州大学精细化工研究开发中心教育部绿色农药与生物工程重点实验室
关键词：
毒氟磷;单克隆抗体;间接竞争酶联免疫分析方法
摘  要：
[目的]以新型抗病毒剂创制品种毒氟磷为研究对象,通过杂交瘤技术建立抗毒氟磷及其类似物的高特异性杂交瘤细胞系,应用体内诱生腹水制备单克隆抗体,开发出检测毒氟磷及其类似物的酶联免疫分析方法。[方法]利用2-甲酰苯氧乙酸、氨基酸合成半抗原,通过碳二亚胺法合成人工抗原,免疫动物并通过杂交瘤技术得到一株杂交瘤细胞株DHS3A2。[结果]在制备的单克隆抗体mAb-DHS3A2的基础上,通过对有机溶剂、pH值、离子强度等因素的筛选比较,优化了毒氟磷间接竞争ic-ELISA,方法的IC50值为(9.62±0.59)μg/L,检出限为(0.3±0.05)μg/L。[结论]该方法快速、准确,适用于不同基质中毒氟磷农药残留快速检测。
译  名：
The Immunoassay Method for the Detection of Dufulin
作  者：
LU Ping;LI Yan-jiao;DING Xiao-yan;Center for Research and Development of Fine Chemicals of Guizhou University, Key Laboratory of Green Pesticide and Agricultural Bioengineering of Education Ministry;
关键词：
dufulin;;monoclonal antibody;;ic-ELISA
摘  要：
[Aims] Dufulin is a novel virus inhibitor which was developed and synthesized by the center for research and development of fine chemicals of guizhou university. The screening detection method must be precise and high sensitive for quantities, as simple and quick and cheap as it possible. The idea of solution of this problem is to develop simple and non-expensive immunoassay methods. [Methods] 6- [2-((Methylbenzothiazol-2-yl)-1-(2-ethoxy)-O,O-diethyl-α-amino-phosphonate)acetamido)] hexanoic acid(DHS) was synthesized. The haptens were then covalently coupled onto keyhole limpet hemocyanin(KLH) or bovine serum albumin(BSA) and served as immunogens(DHS-KLH) and coating antigen(DHS-BSA), respectively. The Balb/c mice immunized five times with DHS-KLH,and the immunized effects were measured by indirect competitive ELISA(ic-ELISA). [Results] Based on the monoclonal antibodies, ic-ELISA conditions such as solvent, pH and ionic strength were optimized. Results showed that the IC50 and LOD of the method was(9.62± 0.59),(0.3± 0.05) μg/L. [Conclusions] The method is fast and accurate to determine dufulin in different samples.

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