Fabrication and Characterization of Monoclonal Antibody Against Isoprothiolane

WANG Xing-xing; WANG Yao-yu; LI Xiao-na; SHEN Chao-qun; ZHOU Kai; ZHANG Jing

doi:10.19969/j.fxcsxb.22042201

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Fabrication and Characterization of Monoclonal Antibody Against Isoprothiolane

Experimental Techniques and Methods | 更新时间：2023-01-04

- Fabrication and Characterization of Monoclonal Antibody Against Isoprothiolane
- Journal of Instrumental Analysis Vol. 41, Issue 11, Pages: 1696-1701(2022)
- 作者机构：
  
  1.深圳市通量检测科技有限公司，广东深圳 518038
  2.九江学院江西油茶研究研究中心，江西九江 332005
  3.烟台海关技术中心，山东烟台 264000
- 作者简介：
- 基金信息：
- DOI：10.19969/j.fxcsxb.22042201
  CLC：
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王幸幸,王耀宇,李晓娜等.稻瘟灵单克隆抗体的制备及评价[J].分析测试学报,2022,41(11):1696-1701.

WANG Xing-xing,WANG Yao-yu,LI Xiao-na,et al.Fabrication and Characterization of Monoclonal Antibody Against Isoprothiolane[J].Journal of Instrumental Analysis,2022,41(11):1696-1701.
王幸幸,王耀宇,李晓娜等.稻瘟灵单克隆抗体的制备及评价[J].分析测试学报,2022,41(11):1696-1701. DOI： 10.19969/j.fxcsxb.22042201.

WANG Xing-xing,WANG Yao-yu,LI Xiao-na,et al.Fabrication and Characterization of Monoclonal Antibody Against Isoprothiolane[J].Journal of Instrumental Analysis,2022,41(11):1696-1701. DOI： 10.19969/j.fxcsxb.22042201.

摘要

该文制备了农药稻瘟灵的单克隆抗体，并建立了稻瘟灵的酶联免疫吸附（ELISA）检测方法。在完全保留稻瘟灵结构的基础上从二硫杂环戊烷结构中衍生不同长度的活性手臂制备了2个半抗原，并分别与载体蛋白偶联合成免疫原与包被原。通过小鼠免疫、细胞融合、淘筛、腹水制备等步骤获得特异性识别稻瘟灵的单克隆抗体mAb－DWL。结果显示，基于mAb－DWL构建的间接竞争ELISA法的半抑制浓度（IC,50,）为55.2 ng/mL，线性范围为4.6 ~ 530.2 ng/mL，其与结构类似物的交叉反应可忽略不计。所建立的ELISA方法对蔬菜及粮食等样品的加标回收率为77.2% ~ 116%，可用于实际样品的快速检测。

Abstract

An enzyme-linked immunosorbent assay（ELISA） method based on the monoclonal antibody was established for determination of pesticide isoprothiolane．Two haptens were prepared by extending active arms of different lengths from disulfide heterocyclic pentane structure of isoprothiolane molecule．The two haptens were conjugated with carrier protein to obtain the immunogen and coating antigen．Monoclonal antibody，named mAb－DWL，against isoprothiolane was obtained via the procedures of mouse immunization，cell fusion，eluting sieve and ascites collection．Results showed that the ELISA based on the obtained mAb－DWL exhibited a linear range of 4.6－530.2 ng/mL，with an IC,50, of 55.2 ng/mL．Meanwhile，the cross-reactivity rates for the mAb－DWL with other similar pesticides were lower than 1%．The spiked recoveries for isoprothiolane in the vegetables and grains samples ranged from 77.2% to 116%．The monoclonal antibody prepared by this method could be used for development of rapid method for isoprothiolane determination.

关键词

稻瘟灵单克隆抗体酶联免疫吸附检测农药

Keywords

isoprothiolanemonoclonal antibodyELISApesticide

references

Li J R，Jiao X Z，Yu L Y．Agrochemicals（李继睿，焦兴忠，禹练英．农药），2010，（11）：798－829.

Che S J．Study on the Sensitivity of Pyricularia Grisea to Isoprothiolane and Tricyclazole in Heilongjiang Province. Heilongjiang：Heilongjiang Bayi Agricultural University（车淑静．黑龙江省稻瘟病菌对稻瘟灵和三环唑的敏感性研究．黑龙江：黑龙江八一农垦大学），2008.

Meng Q S．China Food Addit. （孟庆顺．中国食品添加剂），2020，31（6）：119－122.

Fang X H，Qiu R L．Soil Environ. Sci. （方晓航，仇荣亮．土壤与环境），2002，11（1）：94－97.

GB 2763－2021. National Food Safety Standards. Maximum Residue Limits for Pesticides in Food. National Standards of the People's Republic of China（食品安全国家标准．食品中农药最大残留限量．中华人民共和国国家标准）.

GB 23200.113－2018. National Food Safety Standards. ofDetermination 208 Pesticides and Metabolites Residues in Foods of Plant Origin－Gas Chromatography－Tandem Mass Spectrometry Method. National Standards of the People's Republic of China（食品安全国家标准．植物源性食品中208种农药及其代谢物残留量的测定气相色谱－质谱联用法．中华人民共和国国家标准）.

GB 23200.116－2019. National Food Safety Standards. ofDetermination 90 Organophosphorus Pesticides and Metabolites Residues in Foods of Plant Origin－Gas Chromatography Method. National Standards of the People’s Republic of China（食品安全国家标准．植物源性食品中90种有机磷类农药及其代谢物残留量的测定气相色谱法．中华人民共和国国家标准）.

GB/T 5009.115－2003. National Food Safety Standards. Determination of Tricyclazole Residues in Rice. National Standards of the People's Republic of China（食品安全国家标准．稻谷中三环唑残留量的测定．中华人民共和国国家标准）.

Tang L H．Food Ind. （汤丽华．食品工业），2019，40（2）：303－306.

Qiu W F，Zhang C J，Wen L，Fang Y，Hu Q H．Food Sci. （邱伟芬，张昌娟，文良，方勇，胡秋辉．食品科学），2015，36（12）：221－225.

Wang W J，Ye J，Sun S Y，Wang S X，Sang H C．J. Food Saf. Qual. （王文珺，叶金，孙双艳，王松雪，桑华春．食品安全质量检测学报），2018，9（21）：5552－5558.

Selvi A A，Manonmani H K．J. Immun. Immunochem.，2013，34（2）：149－165.

Estevez M C，Galve R，Sanchez－Baeza F，Marco M P．Anal. Chem.，2005，77（16）：5283－5293.

Fang S，Zhang B，Ren K W，Cao M M，Shi H Y，Wang M H．J. Agric. Food Chem.，2011，59（5）：1594－1597.

Liu B H，Chu K C，Yu F Y．Food Control，2016，66：1－7.

Zhu L，Chen Z J，Luo L，Wang Y，Zhong Y X，Shen Y D，Sun Y M，Xu Z L．J. Instrum. Anal. 朱琳，陈子键，罗林，王宇，钟玉心，沈玉栋，孙远明，徐振林．分析测试学报），2022，41（1）：128－135.

Luo L．Study on the Antibody Production of Three Ultra-low Molecular Organic Contaminants and Development of Immunoassays. Guangzhou：South China Agricultural University（罗林．几种超小分子有害物特异性抗体制备及免疫检测技术研究与应用．广州：华南农业大学），2017.

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