School of Food Science & Technology, State Key Laboratory of Food Science & Technology, Jiangnan University, Wuxi, China.
Immunol Invest. 2013;42(6):493-503. doi: 10.3109/08820139.2013.797909.
In the present study, we obtained a specific monoclonal antibody (cross-reaction to analogues <5%) against cyhalothrin using two haptens. After 7 reaction steps, 3-cyano-[(cis)-3-(2-chloro-3, 3, 3-trifluoroethenyl-2, 2-dimethyl)-cyclopropane-carbonyloxy]-phenoxybenzyl propanoic acid was prepared with yield 35.9%. Four coating antigens and two immunogens were prepared. A heterologous enzyme-linked immunosorbent assay for cyhalothrin was established with the 50% inhibition concentration (IC50, 13.26 ± 1.23 ng mL(-1)) after optimizing various parameters including coating antigens, blocking agents, ionic strength, pH value and methanol concentration in the assay buffer. To evaluate the proposed immunoassay, spiked samples from river, tap water and drinking water at three levels (0.2, 1.0, 5.0 mg L(-1)) were tested after simple dilution. The mean recoveries ranged from 75.4% to 97.7% with coefficient of variation 5.1%-11.6%. The results from the above indicated the potencies of this ELISA in cyhalothrin analysis.
在本研究中，我们使用两种半抗原针对氯氟氰菊酯获得了一种特异性的单克隆抗体（对类似物的交叉反应<5%）。经过 7 步反应，用 35.9%的产率制备了 3-氰基-[(顺式)-3-(2-氯-3,3,3-三氟乙烯基-2,2-二甲基）-环丙烷羰基氧]-苯氧基苄基丙酸。制备了四种包被抗原和两种免疫原。通过优化包括包被抗原、封闭剂、离子强度、pH 值和测定缓冲液中甲醇浓度在内的各种参数，建立了氯氟氰菊酯的异源酶联免疫吸附测定法，其 50%抑制浓度（IC50，13.26±1.23ng mL(-1)）。为了评估所提出的免疫分析方法，在简单稀释后，对来自河流、自来水和饮用水的三个水平（0.2、1.0、5.0mg L(-1)）的加标样品进行了测试。平均回收率在 75.4%至 97.7%之间，变异系数为 5.1%-11.6%。上述结果表明，该 ELISA 在氯氟氰菊酯分析中具有强大的分析能力。
