恩诺沙星单克隆抗体的制备及其鉴定  

Preparation of Monoclonal Antibodies against Enrofloxacin and Identification

作　　者：刘红[1] 曾振灵[2] 杨桂香[2] 陈杖榴[2] 

机构地区：[1]中山市农产品质量监督检验检测中心,广东中山528403 [2]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽药杂志》2006年第10期5-8,共4页Chinese Journal of Veterinary Drug

基　　金：广东省自然科学基金资助项目(994162)

摘　　要：采用碳二亚胺法和氯甲酸异丁酯法合成恩诺沙星的人工抗原,结合比为15∶1。用人工抗原腹腔免疫BALB/c小鼠,有限稀释法筛选获得分泌特异性抗体的杂交瘤细胞株2H10C2F11,并将该细胞株注射小鼠腹腔获得腹水。纯化后的抗体效价为3.96×10-6,紫外分析方法计算蛋白含量为1.139 mg/mL,琼脂双扩实验表明抗体为IgG1型,SDS-PAGE电泳图谱显示纯化效果理想;经间接ELISA方法测定,抗体的亲和力为1.8×108L/moL。Using carbodiimide method and isobutylchloroformate method, two artificial antigens were synthesized. The BALB/c mice were immunized intrapefitoneally with these antigens, and the hybridomas secreting specific antibody 2H10C2F11 was obtained through screened by the finite diluted method. The BALB/c mice were injected intraperitoneally with the 2H10C2F11 for obtaining large quantities of antibody. After purified, the titer of the purified antibody was 3.96 ×10^-6, which is subtype IgG1, and the content of protein was 1. 139 mg/mL. SDS-PAGE analysis of the antibody showed two bands. The affinity of the antibody determined by iELISA was 1.8 ×10^8 L/moL.

关 键 词：恩诺沙星 人工抗原 单克隆抗体 间接ELISA 

分 类 号：Q503[生物学—生物化学]