【摘要】 将多西环素(doxycycline,DOX)进行化学修饰引入羧基或氨基等活性基团,然后与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联合成人工免疫原DOX-PABA-BSA、DOX-BSA和包被原DOX-PABA-OVA、DOX-OVA,并用紫外吸收(UV)、凝胶电泳(SDS-PAGE)和ELISA方法对人工抗原进行鉴定;将合成的人工抗原DOX-PABA-BSA、DOX-BSA分别免疫BALB/C小鼠,用间接ELISA方法测定多抗(pAb)效价,用竞争ELISA方法鉴定其敏感性,用交叉反应试验鉴定其特异性。结果表明,二个免疫组6只小鼠血清抗体效价均在1∶6400以上,DOXpAb对DOX的50%抑制浓度(IC50)在39.79~53.13μg/L,抗血清与四环素类药物交叉反应很低。本实验为建立多西环素ELISA残留免疫学检测方法和并制备多西环素试剂盒奠定了基础。更多还原

【Abstract】 Doxycycline(DOX)was modified with carboxyl or amino-active group,and then with bovine serum albumin(BSA)and ovalbumin(OVA)which were coupled synthesis of artificial immunogen DOX-PABA-BSA,DOX-BSA and coated antigen DOX-PABA-OVA,DOX-OVA.UV Absorption(UV),sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and ELISA were identified on the artificial antigen.The synthesis of artificial antigen DOX-PABA-BSA,DOX-BSA were immunized BALB/C mice.Polyclonal antibody titer were measured by indirect ELISA,using competitive ELISA sensitivity was identified,using cross-reaction tests identify their specificity.The results showed that two immunization groups of six mice with anti-serum titer higher than 1∶6400 by indirect ELISA,50% inhibitive concentration(IC50) was 39.79~53.13 μg/L and low levels of cross-reactivity to Tetracyclines were observed.In this study,In this study,the Pab(anti-DOX antiserum) can be used to develop a ELISA kit to detect DOX residues in animal edible tissues.更多还原