食品科学 ›› 2006, Vol. 27 ›› Issue (10): 223-228.

• 基础研究 • 上一篇    下一篇

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

 于海峰,  张森,  万忠海,  李晓艳, 谭建华   

  1. 吉林大学农学部畜牧兽医学院; 解放军军事医学科学院军事兽医研究所
  • 出版日期:2006-10-15 发布日期:2011-11-16

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

 YU  Hai-Feng,   Zhang-Sen,   Wan-Zhong-Hai,   Li-Xiao-Yan, TAN  Jian-Hua   

  1. 1. College of Animal Science and Technology, Division of Agriculture, Jilin University, Changchun 130062, China;2.Academy of Military Medical Sciences, Veterinarian Research Institute, Changchun 130062, China
  • Online:2006-10-15 Published:2011-11-16

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。

关键词: 氧氟沙星, 单克隆抗体, 酶联免疫吸附试验

Abstract: 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. Mono- clonal 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 ands pecificity.

Key words: Ofloxacin, monoclonal antibodies, ELISA