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可商购的化学物质作为免疫半抗原,用于开发一种可识别多菌灵和其他苯并咪唑类杀真菌剂的多克隆抗体。

Commercially available chemicals as immunizing haptens for the development of a polyclonal antibody recognizing carbendazim and other benzimidazole-type fungicides.

机构信息

Institute of Nuclear and Radiological Sciences & Technology, Energy and Safety (INRaSTES), NCSR "Demokritos", 153 10 Athens, Greece.

Agricultural University of Athens, Faculty of Biotechnology, Iera Odos 75, 118 55 Athens, Greece.

出版信息

Chemosphere. 2015 Jan;119 Suppl:S16-20. doi: 10.1016/j.chemosphere.2014.03.049. Epub 2014 Apr 16.

DOI:10.1016/j.chemosphere.2014.03.049
PMID:24745558
Abstract

Carbendazim is a fungicide widely used for controlling fungi affecting fruits, vegetables, field crops etc. Determination of carbendazim in water, soil and various crops is frequently required to assure compliance with national/European regulations. A polyclonal antibody recognizing carbendazim was developed by using commercially available 2-(2-aminoethyl) benzimidazole, 2-benzimidazole propionic acid and 2-mercaptobenzimidazole as immunizing haptens; each of the above derivatives was directly conjugated to the carrier protein keyhole limpet hemocyanin and a mixture of the conjugates was administered to New Zealand white rabbits. Immunochemical functionality of the antisera and the corresponding isolated antibody (whole IgG fraction) was evaluated through titer and displacement curves in an in-house developed ELISA, which employed a 2-mercaptobenzimidazole - functionalized lysine-dendrimer as the immobilized hapten. As shown with ELISA-displacement curves, the above antibody could recognize carbendazim as well as other benzimidazole-type fungicides, i.e. benomyl and thiabendazole, and also intact benzimidazole, while it did not cross-react with the structurally different pesticides carbaryl and imazalil. Considering the rather simple approach which has led to its development and its highly promising immunochemical profile, the new antibody may be exploited in immunoanalytical systems for detecting benzimidazole-type pesticides e.g. in samples of environmental interest. The above antibody is being currently tested as a biorecognition element in the novel FOODSCAN cell biosensor platform for pesticide residue detection based on the Bioelectric Recognition Assay technology.

摘要

多菌灵是一种广泛用于防治水果、蔬菜、大田作物等真菌的杀菌剂。为了确保符合国家/欧洲法规的要求,经常需要测定水中、土壤中和各种作物中的多菌灵。本研究采用市售的 2-(2-氨基乙基)苯并咪唑、2-苯并咪唑丙酸和 2-巯基苯并咪唑作为免疫原,制备了多菌灵多克隆抗体;上述每种衍生物均直接与载体蛋白血蓝蛋白偶联,然后将混合物免疫新西兰白兔。通过建立的酶联免疫吸附试验(ELISA),用 2-巯基苯并咪唑功能化赖氨酸树枝状聚合物作为固定化半抗原,测定抗体效价和竞争抑制曲线,评估了抗血清和分离的抗体(全 IgG 部分)的免疫化学功能。ELISA 竞争抑制曲线表明,该抗体不仅可以识别多菌灵,还可以识别其他苯并咪唑类杀菌剂,如苯菌灵和噻菌灵,也可以识别完整的苯并咪唑,但与结构不同的农药carbaryl 和 imazalil 无交叉反应。考虑到其开发所采用的相对简单的方法及其非常有前途的免疫化学特性,该新抗体可用于免疫分析系统,检测苯并咪唑类农药,例如环境样品中的农药。目前,该抗体正在基于生物电识别分析技术的新型 FOODSCAN 细胞生物传感器平台农药残留检测中作为生物识别元件进行测试。

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