氧氟沙星完全抗原的合成鉴定及其单克隆抗体的制备纯化  

Synthesis and Identification of the Antigen of OFLX and the Production and Purification of Its Monoclonal Antibodies

作　　者：于海峰[1] 张森[1] 万忠海[2] 李晓艳[2] 谭建华[2] 

机构地区：[1]吉林大学农学部畜牧兽医学院,吉林长春130062 [2]解放军军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《食品科学》2006年第10期223-228,共6页Food Science

摘　　要：本试验通过碳二亚胺(EDC)法把氧氟沙星(OFLX)与载体蛋白BSA和OVA分别进行偶联制备免疫抗原和包被抗原,并采用紫外扫描法和SDS-聚丙烯酰胺凝胶电泳方法对合成抗原进行了鉴定。通过杂交瘤常规技术获得了抗OFLX的单克隆抗体杂交瘤细胞株7株,用辛酸-硫酸铵法对腹水进行了纯化。纯化腹水均属IgG1型。经过ELISA方法鉴定,纯化后的抗体效价>1:128000;交叉实验证明所得抗体与达氟沙星、洛美沙星、环丙沙星、诺氟沙星、卡那霉素、庆大霉素、泰乐菌素均没有交叉反应,表现了良好的敏感性和特异性。The Ofloxacin（OFLX） is coupled to BSA as an immunogen and to OVA as a coating antigen using the coupling agent EDC. The immunogen and coating antigen have been identified by the method of ultraviolet scanner and SDS-PAGE. Monoclonal antibodies hybridoma cell against OFLX 7 strain have been gained and ascites have been purified by using a caprylic acid/ ammonium sulfate precipitation （CA-AS）. Subclasscs of surified ascites was IgG1. Valency of antibody purified is higher than 1：128000, and the antibody wasn＇t cross-reactivity with Danofloxacin, Lomefloxacin, Ciprofloxacin, Norfloxacin, Kanamycin, Gentamicin, Tylosin by ELISA. And the purified monoclonal antibody against OFLX shows a very high degree of sensitivity and specificity.

关 键 词：氧氟沙星 单克隆抗体 酶联免疫吸附试验 

分 类 号：S852.43[农业科学—基础兽医学]