Yang Jinyi, Wang Hong, Jiang Yueming, Sun Yuanming, Pan Ke, Lei Hongtao, Wu Qing, Shen Yudong, Xiao Zhili, Xu Zhenlin
College of Food Science, South China Agricultural University, Guangzhou 510642, PR China.
Molecules. 2008 Apr 17;13(4):871-81. doi: 10.3390/molecules13040871.
The haptens 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)carbonyl]-amino]butanoic acid (BFNB) and 6-[((2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)-carbonylamino]hexanoic acid (BFNH) were synthesized and then used to develop a rapid,specific and sensitive ELISA method to determine residues of the pesticide carbofuran in a variety of matrices. A hybridoma cell line (5D3) producing anti-carbofuran monoclonal antibodies (MAbs) was also established. Based on the MAbs in combination with the heterologous hapten BFNH coupled to either horseradish peroxidase (HRP) or ovalbumin(OVA), four ELISAs (formats I-IV) for the quantification of carbofuran were developed and compared. Among them, the optimized format II (the conjugate-coated direct competitive ELISA) showed the best characteristics, with an IC50 value of 18.49 ng/mL, a limit of detection of 0.11 ng/mL and the shortest assay time (1 h). This ELISA method was then applied to the determinations of carbofuran in environmental water, soil and food samples. The relative standard deviations (R.S.D.s) ranged from 1.8% to 21.3% and the mean recoveries were 104.6%, 108.3%, 106.3% and 100.1% for water, soil, lettuce and cabbage, respectively. Thus, the ELISA method of format II exhibited the potential to develop commercial ELISA kits for a rapid detection of carbofuran for human health and environmental safety.
合成了半抗原4-[[(2,3-二氢-2,2-二甲基-7-苯并呋喃氧基)羰基]-氨基]丁酸（BFNB）和6-[((2,3-二氢-2,2-二甲基-7-苯并呋喃氧基)羰基氨基]己酸（BFNH），然后用它们开发了一种快速、特异且灵敏的酶联免疫吸附测定（ELISA）方法，用于测定多种基质中农药克百威的残留量。还建立了产生抗克百威单克隆抗体（MAbs）的杂交瘤细胞系（5D3）。基于这些单克隆抗体，结合与辣根过氧化物酶（HRP）或卵清蛋白（OVA）偶联的异源半抗原BFNH，开发并比较了四种用于定量克百威的ELISA方法（形式I-IV）。其中，优化后的形式II（偶联物包被直接竞争ELISA）表现出最佳特性，半数抑制浓度（IC50）值为18.49 ng/mL，检测限为0.11 ng/mL，且检测时间最短（1小时）。然后将该ELISA方法应用于环境水、土壤和食品样品中克百威的测定。水、土壤、生菜和卷心菜的相对标准偏差（R.S.D.s）范围为1.8%至21.3%，平均回收率分别为104.6%、108.3%、106.3%和100.1%。因此，形式II的ELISA方法具有开发用于快速检测克百威的商业ELISA试剂盒以保障人类健康和环境安全的潜力。
