抗恩诺沙星单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibodies against Enrofloxacin

作　　者：张家禾[1] 周作红[1] 洪伟鸣 刘莉 孟婷 王永娟 管远红 左伟勇 

机构地区：[1]江西农业大学动物科技学院,江西南昌330045 [2]江苏省兽用生物制药高技术研究重点实验室,江苏泰州225300

出　　处：《江西农业大学学报》2013年第5期1070-1073,共4页Acta Agriculturae Universitatis Jiangxiensis

基　　金：安徽省科技攻关项目(1201c0602006);江苏省"青蓝工程"中青年学术带头人项目资助

摘　　要：制备抗恩诺沙星的单克隆抗体并对其生物学特性进行鉴定。采用活化酯法合成恩诺沙星-牛血清白蛋白作为免疫原免疫小鼠,利用杂交瘤细胞技术建立杂交瘤细胞株,利用体内诱生法制备单克隆抗体,然后对制备的单抗的效价、敏感性、特异性、亲和力和亚型等免疫学特性进行鉴定。获得细胞株7C5,对单抗亚型鉴定结果均为IgG1,诱生腹水中抗体的效价在1.035×10-6以上,最低检出限为0.56 ng/mL,IC50值为14.21 ng/mL,检测范围为2.05～98.38 ng/mL,与其他同类药物无交叉反应。试验所获得抗体效价高、特异性强,可以为ELISA试剂盒和胶体金试纸条研发提供良好的抗体材料。Preparation and identification of monoclonal antibodies （mAb） against Enrofloxacin （EFLX） Immunogen were carried out.EFLX-BSA was synthesized using the activated ester method,the hybridoma cell strain secreting anti-EFLX mAb was obtained by using hybridoma technology.The massive mAb was induced with in vivo method,the titer,affinity,sensitivity,specificity and subtype of the mAb were characterized.The hybridoma cell lines of 7C5 were screened for specificity to EFLX,the isotypes of the mAb were IgG1.The indirect ELISA titer of the mAb was 1.035 ×10-6 in supernatant.The result had a linear detection range from 2.05 ng/mL to 98.38 ng/mL （R2 =0.992 7）,with IC50 and LOD values of 14.21 ng/mL and 0.56 ng/mL,the mAb had no cross reaction to other antibiotics.The antibody obtained in this experiment is of high utility and strong specificity,which may provide good antibody materials for the research and development of colloidal gold test and ELISA kit.

关 键 词：恩诺沙星 杂交瘤技术 单克隆抗体 交叉反应 

分 类 号：Q785[生物学—分子生物学]