本研究合成并鉴定了黄曲霉毒素M1人工抗原，通过动物免疫制备出敏感性高、特异性好的鼠源黄曲霉毒素M1多克隆抗血清。采用琥珀酸酐改造黄曲霉毒素M1，并将其改造物按两种方法(DCC和EDC)分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)进行偶联，合成免疫抗原AFM1-BSA和检测抗原AFM1-OVA，经过薄层色谱、紫外扫描和凝胶电泳鉴定后，免疫BALB/c小鼠，共免疫3次，每次间隔3周，最后1次免疫10d后，断尾采血，制备多抗血清。经过测定，免疫的3只小鼠效价均达到了1：104以上，2号小鼠多抗血清的敏感性最好，半数抑制浓度IC50（50% inhibitive concentration，IC50）为37.61ng/mL，且具有良好的特异性。本研究成功合成了黄曲霉毒素M1人工抗原和多克隆抗体血清，为黄曲霉毒素M1单克隆抗体制备及其免疫学快速检测方法的建立奠定了基础。%To prepare high sensibility and speciifcity polyclonal antiserum against AFM1, AFM1 artiifcial antigens were synthesized and identified through animal immune in this study. Firstly AFM1 was modified by succinic anhydride method and connected to carrier protein via active ester method (DCC) and Carbodiimide method (EDC). Two complete antigens of AFM1 (AFM1-BSA and AFM1-OVA) were prepared successfully. After the identiifcation test via TLC method, UV spectrophotometry and SDS-PAGE, synthetic antigens were injected into experimental animals(BALB/c mouse), once every three weeks for 3 times, ten days after last immunity, poly-antiserum was prepared by broken tail. The electrophoretic velocity of AFM1-BSA was less than that of BSA. All the titers of three immunized mice were 1×10-4 approximately, the multi-antiserum from No.2 sample had the best sensitivity, its IC50 was 37.61ng/mL and has good speciifcity. In this study, we obtained AFM1 artiifcial antigen and murine multi-antiserum of high sensitivity, which are preparing for the monoclonal antibody of AFM1 and the research of rapid analysis kits or strips.
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