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| 禽流感免疫噬菌体抗体库构建及ELISA检测方法的建立 | |
| 引用本文： | 邢佑尚,汪琳,张灿,赵胤泽,栢亚铎,吴亚琼,李玉欣.禽流感免疫噬菌体抗体库构建及ELISA检测方法的建立[J].中国畜牧兽医,2012,39(6):17-21. |
| 作者姓名： | 邢佑尚 汪琳 张灿 赵胤泽 栢亚铎 吴亚琼 李玉欣 |
| 作者单位： | 1. 沈阳农业大学,辽宁沈阳110886;北京出入境检验检疫局,北京100026 2. 北京出入境检验检疫局,北京,100026 3. 军事医学科学院卫生学环境医学研究所,天津,300050 4. 西南大学,重庆,400716 |
| 基金项目： | 国家质检总局科技计划项目 |
| 摘 要： | 本研究应用噬菌体抗体库技术筛选禽流感亚型病毒Fab抗体,并建立相应ELISA检测方法。用禽流感亚型混合疫苗免疫小鼠,取其脾脏提取总RNA用于扩增免疫球蛋白轻链和重链基因,克隆至噬菌粒pComb3,然后转化入感受态细胞XL1-Blue中,以1012 CFU辅助噬菌体VCSM13进行感染后,获得Fab抗体库容量为8×107 CFU。以H5N1病毒为抗原进行4轮亲和筛选,获得特异性结合的克隆,利用获得的特异性克隆进行ELISA检测方法的建立。 |
| 关 键 词： | 禽流感病毒 噬菌体抗体库 检测方法建立 |
| 收稿时间： | 2012-02-27 |
Construction of Avian Influenza Immune Phage Antibody Library and Establishment of an ELISA Detection Method |
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| XING You-shang , WANG Lin , ZHANG Can , ZHAO Yin-ze , BAI Ya-duo , WU Ya-qiong , LI Yu-xin.Construction of Avian Influenza Immune Phage Antibody Library and Establishment of an ELISA Detection Method[J].China Animal Husbandry & Veterinary Medicine,2012,39(6):17-21. | |
| Authors: | XING You-shang WANG Lin ZHANG Can ZHAO Yin-ze BAI Ya-duo WU Ya-qiong LI Yu-xin |
| Affiliation: | 1. Shenyang Agricultural University,Shenyang 110886,China;2. Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing 100026,China;3. Institute of Hygiene and Environmental Medicine, Academy of Military Medical Sciences, Tianjin 300050, China;4. Southwest University,Chongqing 400716,China |
| Abstract: | Fab antibody of avian influenza virus subtypes was screened by phage antibody library,and an ELISA detection method was established. Mice were immunized with vaccine mixed of subtypes of AIV, the total RNA was extracted from harvested spleens, immunoglobulin light-chain and heavy-chain gene was amplificated with the template of total RNA, and cloned into phagemid pComb3, the recombinant phagemid pComb3 was transformed into XL1-Blue competent cells, after infected with helper phage VCSM13, Fab antibody library of 8?107 CFU was obtained. Specific clones anti-H5N1 virus was obtained after 4 rounds of screening,ELISA detection method was established with specific clones. |
| Keywords: | avian influenza virus phage antibody library establishment of detection method |
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