金霉素单克隆抗体的制备及检测方法的建立  

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

作　　者：张云涛[1] 路东[1] 席仲兴[1] 张正雷[1] 曹馨匀[1] 谢雍[2] 

机构地区：[1]兰州生物制品研究所,兰州730046 [2]香港科技大学

出　　处：《微生物学免疫学进展》2008年第4期55-59,共5页Progress In Microbiology and Immunology

基　　金：甘肃省重大科技专项(0702NKDA043)

摘　　要：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。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 es- tablished 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 mono- clonal antibodies. As little as 50ng of AM per milliliter can be detected. The kits don＇ t cross- react with other antibiotic.

关 键 词：金霉素 羰基二咪唑 杂交瘤 单克隆抗体 酶联免疫吸附试验 

分 类 号：S859.84[农业科学—临床兽医学]