INFLUENCE OF THE ELISA FORMAT AND THE HAPTEN ENZYME CONJUGATE ON THE SENSITIVITY OF AN IMMUNOASSAY FOR S-TRIAZINE HERBICIDES USING MONOCLONAL-ANTIBODIES
INFLUENCE OF THE ELISA FORMAT AND THE HAPTEN ENZYME CONJUGATE ON THE SENSITIVITY OF AN IMMUNOASSAY FOR S-TRIAZINE HERBICIDES USING MONOCLONAL-ANTIBODIES
复制标题
DOI：
10.1021/jf00015a033
复制
发表时间：
1992-03-01
影响因子：
6.1
通讯作者：
HAMMOCK, BD
中科院分区：
文献类型：
--
作者：
SCHNEIDER, P;HAMMOCK, BD
各种形式的酶联免疫吸附测定已被检查的环境样品的分析使用一个单一的单克隆抗体提出的除草剂阿特拉津。几种ELISA形式和各种合成的半抗原-酶缀合物的最有利的组合导致在0.25 μ g/L下具有50%抑制（I50）和0.03 μ g/L的检测限的高度灵敏的酶免疫测定。该检测是最灵敏的阿特拉津检测，基于迄今为止发表的单克隆抗体，可在1小时内完成。所有形式均基于半抗原-酶缀合物（酶示踪剂）的使用。使用直接包被和用山羊抗小鼠抗体或蛋白A预包被。使用预包被步骤的形式被证明是最灵敏的。使用碱性磷酸酶或辣根过氧化物酶缀合基于两种不同的阿特拉津和西玛津半抗原的酶示踪剂，每种半抗原具有两个不同的间隔区（C3或C6）。具有与辣根过氧化物酶缀合的C6间隔臂的阿特拉津衍生物提供了最高的灵敏度。
A variety of formats for enzyme-linked immunosorbent assays have been examined for the analysis of environmental samples using a single monoclonal antibody raised to the herbicide atrazine. The most favorable combination of several ELISA formats and a variety of synthesized hapten-enzyme conjugates resulted in a highly sensitive enzyme immunoassay with 50% inhibition (I50) at 0.25-mu-g/L and a detection limit of 0.03-mu-g/L. This assay is the most sensitive atrazine assay, based on monoclonal antibodies published so far, and can be performed in less than 1 h. All formats were based on the use of hapten-enzyme conjugates (enzyme tracers). Direct coating and precoating with goat anti-mouse antibodies or protein A were used. The formats using precoating steps turned out to be the most sensitive, Enzyme tracers based on two different atrazine and simazine haptens each with two different spacers (C3 or C6) were conjugated using either alkaline phosphatase or horseradish peroxidase. The atrazine derivative with the C6 spacer arm conjugated to horseradish peroxidase provided highest sensitivity.