| 抗庆大霉素噬菌体抗体库的构建和筛选 |
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| 引用本文： | 孟辉,温凯,沈建忠,王战辉,张素霞.抗庆大霉素噬菌体抗体库的构建和筛选[J].中国畜牧兽医,2011,38(1):76-80. |
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| 作者姓名： | 孟辉 温凯 沈建忠 王战辉 张素霞 |
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| 作者单位： | （中国农业大学动物医学院，北京 100193） |
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| 基金项目： | 国家高技术研究发展计划(863计划)(2008AA10Z423); 国家自然科学基金重点项目(30830082) |
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| 摘 要： | 本研究使用噬菌体抗体库技术筛选针对庆大霉素（gentamicin）的单链抗体。以抗庆大霉素杂交瘤细胞株（2A3）为基因来源构建scFv基因，连接至pCANTAB5E噬菌粒载体，通过电击转化TG1（Escherichia coli TG1）构建了库容量为6.5×106噬菌体单链抗体库。抗庆大霉素噬菌体单链抗体库进行三轮富集筛选，通过Phage-ELISA技术成功筛选出了9个抗庆大霉素噬菌体阳性克隆，为开发新型残留检测用抗体奠定了基础。
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| 关 键 词： | 庆大霉素 噬菌体抗体库 单链抗体 |
Construction of Anti-gentamicin Phage Display scFv Libraries, Selection and Identification |
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| MENG Hui,WEN Kai,SHEN Jian-zhong,WANG Zhan-hui,ZHANG Su-xia.Construction of Anti-gentamicin Phage Display scFv Libraries, Selection and Identification[J].China Animal Husbandry & Veterinary Medicine,2011,38(1):76-80. |
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| Authors: | MENG Hui WEN Kai SHEN Jian-zhong WANG Zhan-hui ZHANG Su-xia |
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| Affiliation: | (College of Veterinary Medicine，China Agricultural University，Beijing 100193，China) |
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| Abstract: | In this study, we constructed a phage display antibody library specific for gentamicin (GEN) to get the anti-getamicin single chain antibody(scFv). We used the anti-gentamicin hybridoma (2A3) as a source of gene to produce full-length scFv fragment. The full-length scFv fragment was ligated into the phagemid vector pCANTAB5E. Then the phagemid was transformed to E.coli TGl cells by electroporation. The size of antibody libraries was 6.5×106. The primary phage display library was riched by stabilized antigen immunoaffinity panning with GEN-OVA conjugates. 9 clones congtained insert and showed GEN-positive in phage-ELISA, which lay the foundation for further immunological analysis. |
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| Keywords: | gentamicin phage display antibody library single chain antibody |
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