| G族黄曲霉毒素半抗原分子设计、抗原合成及抗体特性 |
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| 引用本文：王亚楠,王晓斐,牛琳琳,雷 壮,张海棠,王自良.G族黄曲霉毒素半抗原分子设计、抗原合成及抗体特性[J].西北农业学报,2018,27(5):626~633 |
| DOI:10.7606/j.issn.1004-1389.2018.05.003 |
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| 基金项目:“十二五”国家科技支撑计划(2014BAD13B05)。 |
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| 中文摘要:根据黄曲霉毒素G1（Aflatoxin G1，AFG1）的分子结构和活性位点，采用半缩醛法（SA）、环氧化物法（EP）和烯醇醚衍生物法（EED）制备G族AF人工抗原AFG1-BSA，通过UV和SDS-PAGE进行鉴定。用AFG1-BSA免疫Balb/C小鼠制备多克隆抗体（AFG1 pAb），间接ELISA检测AFG1 pAb效价，阻断ELISA分析其敏感性，交叉反应试验分析其特异性。结果显示，AFG1-BSA合成成功， 3种合成方法中，SA法效果最好，AFG1与BSA的分子结合比为4.32∶1，间接ELISA效价为1∶（6.4×10），阻断ELISA检测其IC50为13.6 μg·kg-1，与AFG2的交叉反应率（CR）为82.19%，与AFB1、B2无CR。表明获得高效价、敏感、特异、广谱的AFG pAb，为G族AF和AF总量免疫检测方法的建立奠定基础。 |
| 中文关键词:G族黄曲霉毒素 半抗原 人工抗原 多克隆抗体 特性分析 |
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| Hapten Molecular Design and Antigen Synthesis and Antibody Characterization of G-group Aflatoxins |
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| Abstract:According to molecular structure and active site of aflatoxin G1 (AFG1),the G-group AF artificial antigen AFG1-BSA was prepared by using three methods SA,EP and EED and identified by UV and SDS-PAGE.Polyclonal antibodies against AFG1(AFG1 pAb) were prepared by immunizing Balb/C mice with AFG1-BSA,and the titers of AFG1 pAb was detected by indirect ELISA,the sensitivity of AFG1 pAb was analyzed by blocking ELISA and the specificity of AFG1 pAb was analyzed by cross reactivity(CR) test.The results showed that AFG1-BSA was synthesized successfully and the best one was SA method among three synthesis methods of G-group AF artificial antigen and its conjugation ratio of AFG1 to BSA was about 4.32∶1.The immune efficacy of SA method was the best,its AFG1 pAb had high titers of 1∶（6.4×10） by indirect ELISA,a good sensitivity with the 50% inhibition concentration(IC50) of 13.6 μg·kg-1 to AFG1 by blocking ELISA and a high CR to AFG2 of 82.19%,little or no CR to AFB1 and B2.The high-titer,sensitive,specific and broad G-group pAb were produced in sera of immunized Balb/C mice,which it was possible to establish immunoassay of G-group AF and total AF residues in food and agricultural products. |
| keywords:G-group aflatoxins Hapten Artificial antigen Polyclonal antibody Specificity analysis |
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