Development of an enzyme-linked immunosorbent assay for toosendanin

文献类型： 外文期刊

第一作者： Zhang, Jing

作者： Zhang, Jing;Feng, Gang;Luo, Li;Ma, Zhi Qing;Feng, Jun Tao;Zhang, Xing;Yu, Xiang Yang;Liu, Xian Jin

作者机构：

关键词： Toosendanin;Hapten;Polyclonal antibody;Enzyme-linked immunosorbent;assay

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.558； 五年影响因子：6.228 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Enzyme-linked immunosorbent assays(ELISAs)were developed by using polyclonal antibody for toosendanin(TSN),a biopesticide from Melai toosendan Sieb.et Zucc.Their application in the determination of this analyte in spiked cabbage,tomato and apple samples was studied.The haptens,28-hemisuccinyl-TSN(TSN-S)and 28-hemiglutaryl-TSN(TSN-G)were synthesized by using esterification.Immunogen and coating antigen were synthesized by using the mixed anhydride reaction and active ester protocol,respectively.Rabbits were immunized with TSN-G-BSA and TSN-S-BSA.Using the selected antibody and coating antigen,an indirect competitive ELISA for TSN was developed,which showed an IC50 value of 1.023 mu g mL~(-1),with a detection limit of 0.009 mu gmL~(-1).A direct competitive ELISA using an enzyme tracer was also developed.The assay showed an IC50 value of 0.840 mu g mL~(-1)with a detection limit of 0.014 mu gmL~(-1).Both assays displayed high cross-reactivity to a closely structurally related compound.Recoveries of TSN from both immunoassays of fortified samples ranged from 76.4% to 113.2% and 75.1% to 132.3%,respectively.Linear regression analysis showed good correlation between the TSN concentrations derived from ELISA and HPLC analyses,which suggested that the ELISA is a convenient supplementary analytical tool for monitoring TSN.

分类号： O65

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