金霉素单克隆抗体的制备及检测方法的建立

采用羰基二咪唑法,将半抗原金霉素(AM)分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联制备金霉素免疫抗原AM-BSA和检测抗原AM-OVA,通过紫外光谱扫描检测偶联产物。采用细胞杂交瘤技术,制备抗金霉素单克隆抗体杂交瘤细胞株,建立了金霉素竞争ELISA检测方法,其灵敏度达到50ng/ml,且呈现良好的线性关系(r=0.9812),并且与其他抗生素无交叉反应。

Preparation of monoclonal antibody and establishment of detection method by ELISA for aureomycin

In order to establish an enzyme-linked immunosorbent assay(ELISA) for rapidly examing residual aureomycin(AM) in food,the antibody with high titer and specificity must be firstly prepared.It was necessary for antibody production to conjugate the hapten(AM) with a carrier protein to synthesize immunogen(antigen).AM was coupled with carrier protein bovine serum albumin(BSA) and ovdbumin(OVA) to prepare complete antigen AM-BSA and AM-OVA by N,N′-carbonyldiimidazole(CDI).The successful linkage of the AM-BSA was identified by UV.Hybridoma cell lines were established by the hybridoma technique based on the BALB/c mice,which were immunized with AM-BSA.All the results indicated that the immunogen AM-BSA was successfully synthesized.The monoclonal antibodies producted were stable and specific.The kits for measurement of aureomycin concentrations were developed with competitive ELISA based on monoclonal antibodies.As little as 50ng of AM per milliliter can be detected.The kits don't cross-react with other antibiotic.