吖啶酯偶联单克隆抗体建立化学发光法检测黄曲霉毒素B1

doi:10.12160/j.issn.1672-5190.2020.06.015

畜牧与饲料科学 ›› 2020, Vol. 41 ›› Issue (6): 78-83.doi: 10.12160/j.issn.1672-5190.2020.06.015

吖啶酯偶联单克隆抗体建立化学发光法检测黄曲霉毒素B₁

裘雪梅, 朱立鑫, 杨妮, 刘云祎, 刘仁荣

江西科技师范大学生命科学学院,江西南昌 330013

收稿日期:2020-04-08 出版日期:2020-11-30 发布日期:2020-12-17
通讯作者: 刘仁荣（1969—）,男,教授,博士,主要从事食品安全检测研究工作。
作者简介:裘雪梅（1981—）,女,讲师,硕士,主要从事食品安全检测研究工作。
基金资助:
国家自然科学基金项目“基于荧光标记模拟肽内标的黄曲霉毒素柱容量实时监测IAC-HPLC分析方法研究”（31960501）

Detection of Aflatoxin B₁ with the Chemiluminescence Method Established by Acridinium Ester Coupling with Monoclonal Antibody

QIU Xue-mei, ZHU Li-xin, YANG Ni, LIU Yun-yi, LIU Ren-rong

The Life Science College,Jiangxi Science and Technology Normal University,Nanchang 330013,China

Received:2020-04-08 Online:2020-11-30 Published:2020-12-17

摘要/Abstract

摘要： [目的] 建立快速检测黄曲霉毒素B₁（aflatoxin B₁,AFB₁）的方法。[方法] 将吖啶酯（acridinium ester,AE）与AFB₁单克隆抗体进行体外化学合成,分别合成了吖啶酯∶AFB₁单克隆抗体摩尔比为5∶1、10∶1、15∶1、20∶1、25∶1的吖啶酯标记AFB₁单克隆抗体（AFB₁-AE）,分析了不同摩尔比的检测效果,选择吖啶酯∶AFB₁单克隆抗体为25∶1的AFB₁-AE偶联物建立化学发光免疫分析法（chemiluminescence immunoassay,CLIA）。[结果] 获得对应的标准曲线回归方程为：y=-0.245Ln（x）+ 1.578 1,R²=0.985 7,IC₅₀=81.48 pg/mL;对样品进行加标回收实验,加标回收率88.4%~106.0%,变异系数0.92%~2.10%。[结论] 将吖啶酯与AFB₁单克隆抗体体外交联,可建立新的化学发光免疫分析方法。

关键词: 黄曲霉毒素B₁, 吖啶酯, 化学发光免疫分析法, 偶联

Abstract: [Objective] To establish a rapid detection method for aflatoxin B₁ （AFB₁）. [Method] The acridinium ester （AE） and AFB₁ monoclonal antibody were chemically synthesized in vitro, and the AE labeled AFB₁ monoclonal antibody （AFB₁-AE） with different molar ratios of AE and AFB₁ monoclonal antibody of 5∶1, 10∶1, 15∶1, 20∶1 and 25∶1 was synthesized, respectively; the detection effects of AFB₁-AE conjugates with different molar ratios were analyzed, and the conjugate with molar ratio of 25∶1 of AE and AFB₁ monoclonal antibody was finally selected to establish chemiluminescence immunoassay （CLIA）. [Result] The corresponding regression line equation was obtained: y=-0.245Ln（x） + 1.578 1, R²=0.985 7, IC₅₀=81.48 pg/mL; the sample recovery experiment was carried out, and the standard recovery rate was 88.4%-106.0%, the coefficient of variation was 0.92%-2.10%. [Conclusion] In vitro cross-linking of AE with AFB₁ monoclonal antibody can be used to establish a new chemiluminescence immunoassay method.

Key words: aflatoxin B₁, acridinium ester, chemiluminescence immunoassay, coupling

中图分类号:

裘雪梅, 朱立鑫, 杨妮, 刘云祎, 刘仁荣. 吖啶酯偶联单克隆抗体建立化学发光法检测黄曲霉毒素B₁[J]. 畜牧与饲料科学, 2020, 41(6): 78-83.

QIU Xue-mei, ZHU Li-xin, YANG Ni, LIU Yun-yi, LIU Ren-rong. Detection of Aflatoxin B₁ with the Chemiluminescence Method Established by Acridinium Ester Coupling with Monoclonal Antibody[J]. Animal Husbandry and Feed Science, 2020, 41(6): 78-83.

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吖啶酯偶联单克隆抗体建立化学发光法检测黄曲霉毒素B₁

Detection of Aflatoxin B₁ with the Chemiluminescence Method Established by Acridinium Ester Coupling with Monoclonal Antibody

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