伏马菌素B1人工抗原的合成及鼠源多克隆抗血清的制备

本研究合成并鉴定伏马菌素B₁(FB₁)人工抗原,通过动物免疫制备敏感性高、特异性好的鼠源FB₁多克隆抗血清。采用碳二亚胺(EDC)法将FB₁分别与载体蛋白BSA和OVA偶联,合成免疫抗原FB₁-BSA和检测抗原FB₁-OVA,经鉴定后,分别按10和30μg/只的剂量免疫BALB/c小鼠,共免疫4次,每次间隔3周,最后1次免疫30d后,断尾采血,制备多抗血清。利用间接ELISA方法测定抗血清效价,间接竞争ELISA测定敏感性和特异性。结果表明,免疫的6只小鼠效价均达1∶104以上,3号小鼠多抗血清敏感性最好,半数抑制浓度IC₅₀为61.3ng/ml,且具有良好的特异性。本试验成功合成了FB₁人工抗原,并制备了敏感性高、特异性好的鼠源多克隆抗体血清,为FB₁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。

PREPARATION OF FUMONISIN B1 ARTIFICIAL ANTIGEN AND MOUSE POLYCLONAL ANTISERUM

To prepare high sensibility and specificity polyclonal antiserum against FB1,Fumonisin B1(FB1) artificial antigens were synthesized and identified through animal immune in this study.FB1 were separately coupled with carrier protein BSA and OVA using EDC method,then the immune antigen FB1-BSA and detection antigen FB1-OVA were obtained.The BALB/c mice were respectively immune at the dose of 10 and 30μg,every four weeks,four times in total.At the thirty days after the last immunity,the poly-antiserum was prepared through broken tail method.The titer of antiserum was determined by indirect ELISA,sensibility and specificity was determined by indirect competitive ELISA,respectively.The results showed that the high titer of the antiserum of six mice were above 1∶104.The antiserum of No.3 mouse has the best sensibility.Its IC50 was 61.3ng/ml and has good specificity.In this study,FB1 artificial antigens were successful synthesized.FB1 poly-antiserum with high sensibility and specificity were obtained,and this poly-antiserum may lay a foundation for the further study about FB1 monoclonal antibody preparation and the immunology fast detection methods.