| 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 |
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| 引用本文： | 于海峰,张森,万忠海,李晓艳,谭建华. 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[J]. 食品科学, 2006, 27(10): 223-228 |
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| 作者姓名： | 于海峰 张森 万忠海 李晓艳 谭建华 |
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| 作者单位： | 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 |
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| 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。
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| 关 键 词： | 氧氟沙星 单克隆抗体 酶联免疫吸附试验 |
| 文章编号： | 1002-6630（2006）10-0223-06 |
| 收稿时间： | 2006-06-29 |
| 修稿时间： | 2006-06-29 |
Synthesis and Identification of the Antigen of OFLX and the Production and Purification of Its Monoclonal Antibodies |
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| YU Hai-feng,ZHANG Sen,WAN Zhong-hai,LI Xiao-yan,TAN Jian-hua. Synthesis and Identification of the Antigen of OFLX and the Production and Purification of Its Monoclonal Antibodies[J]. Food Science, 2006, 27(10): 223-228 |
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| Authors: | YU Hai-feng ZHANG Sen WAN Zhong-hai LI Xiao-yan TAN Jian-hua |
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| Affiliation: | 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 |
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| 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. |
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| Keywords: | Ofloxacin monoclonal antibodies ELISA |
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