脱氧雪腐镰刀菌烯醇七肽模拟表位淘选及应用

目的从噬菌体随机七肽库中淘选脱氧雪腐镰刀菌烯醇（DON）的模似表位，并以之替代人工抗原建立ELISA检测方法。方法利用抗脱氧雪腐镰刀菌烯醇单克隆抗体12D1，从噬菌体随机七肽库中淘选到脱氧雪腐镰刀菌烯醇七肽模似表位，七肽模拟表位氨基酸序列为CMRPWLQ。以含有DON七肽模拟表位的噬菌体替代DON人工抗原建立DON间接竞争ELISA检测方法，检测范围为20-400ng／ml，并应用于小麦、玉米样品中DON含量的检测。结果检删结果与德国r—Biopharm公司DON检测试剂盒差异无统计学意义。结论DON噬菌体模拟表位替代DON人工抗原建立的ELISA检测方法可应用于小麦和玉米样品中DON的检测。

Identification and application of 7 peptide mimotope which binded to monoclonal antibody of anti - deoxynivalenol

Objective To select phage-displayed mimotipe peptides that mimic the binding of deoxynivalenol(DON) to monoclonal antibodies(McAb) anti-DON,and using the DON mimotope peptide which instead of toxic DON artificial antigen;to establish ELISA method for detecting deoxynivalenol in food products.Methods Phage clones were obtainned by panning from Ph D.-7 Phage Display Peptide Library,producing phage that bound specifically to monoclonal antibody 12DI anti-DON.The amino acid sequence of the binding peptide was CMRPWLQ.A phage indirect competitive enzyme-linked immunosorbent assay(named phage-ELISA) was established by using the mimotope phage in stead of DON artificial antigen for assaying deoxynivalenol concentration in corn and wheat.The linear range of the phage-ELISA was between 20?ng/ml and 400?ng/ml.Results DON concentration in wheat and corn samples detected by phage-ELISA were consistent with the results by the DON kit from r-Biopharm Company.Conclusion Phage mimotope peptide of deoxynivalenoi are effective substitutes for deoxynivalenol to establish ELISA procedures for detecting deoxynivlenol in wheat and corn.