改良重氮法合成米诺环素人工抗原及结果鉴定

The Synthesis and Identification of Minocycline Artificial Antigen Prepared by Improved Diazotization Method

分别采用重氮法、戊二醛法及改良重氮法将米诺环素半抗原与牛血清白蛋白(BSA)偶联制备免疫原,3种方法的偶联比分别为5∶1,10∶1和15∶1;紫外光谱、红外光谱、凝胶电泳对改良重氮法合成的人工抗原进行鉴定,成功合成了人工抗原;将改良重氮法合成的人工抗原MNC-BSA免疫BALB/C小鼠,用间接ELISA法测定抗体效价及其特异性。结果表明:小鼠血清抗体效价均在1∶12800以上,为研究制备米诺环素单克隆抗体奠定了基础。

Minocycline（MNC） was coupled with bovine serum albumin（BSA） to prepare the artificial antigen of MNC-BSA by diazotization method,glutaraldehyde method and improved diazotization method,.respectively.The coupling ratio was 5∶1,10∶1and 15∶1,respectively.The results of ultraviolet spectrum,infrared spectrum and gel electrophoresis identification showed that the artificial antigen of minocycline were synthesizd successfully by the improved diazotization method.The BALB/C mice were immunized by the artificial antigen of MNC-BSA prepared.The result of indirect ELISA showed that the antibody titer for all mice tested were higher than 12800.The result in paper laid the foundations for preparating monoclonal antibody of MNC.