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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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细交链孢菌酮酸酶联免疫吸附分析方法研究
DOI: 10.3724/SP.J.1096.2012.11253, PP. 1347-1352
Keywords: 细交链孢菌酮酸,异源包被,半抗原,多克隆抗体,免疫分析
Abstract:
采用水合肼和乙醛酸依次对细交链孢菌酮酸(Tenuazonicacid,TeA)进行衍生化,设计合成了含有氮杂共轭双键偶联手臂,可增强免疫效果的半抗原TeAHGA。通过偶联载体蛋白BSA后的免疫原TeAHGA-BSA免疫新西兰大白兔,成功制备了特异性识别TeA水合肼衍生物TeAH的多克隆抗体;优化确立了ELISA最佳反应条件(TeAH-OVA为异源包被原、包被浓度0.156μg/L、药物稀释及反应缓冲液为PBS、一抗反应时间40min、二抗反应时间20min),建立了TeA间接竞争ELISA(icELISA)检测方法,其抑制中浓度(IC50)为1.61μg/L,检出限(LOD)为0.08μg/L,定量线性检测范围为0.19-12.89μg/L(IC20-IC80)。番茄、面粉样品平均添加回收率分别为67.2%-89.8%和74.8%-93.7%。
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