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人源性噬菌体抗体库的构建及抗HFRS病毒抗体的筛选

Construction of phage library of human antibody and selection of antibodies against HFRS virus

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【作者】 吴兴安; 徐志凯; 姜绍谆; 阎岩; 白文涛; 王海涛; 张芳琳; 薛小平;

【Author】 WU Xing An, XU Zhi Kai, JIANG Shao Zhun,YAN Yan, BAI Wen Tao, WANG Hai Tao, ZHANG Fang Lin, XUE Xiao Ping Department of Microbiology, Faculty of Preclinical Medicine, Fourth Military Medical University, Xi’an 710033, China

【机构】 第四军医大学基础部微生物学教研室!陕西西安710033; 第四军医大学基?;

【摘要】 目的:构建人源性 Fab 噬菌体抗体基因库并筛选可表达与肾综合征出血热( H F R S)病毒抗原特异结合之 Fab 抗体的重组克隆. 方法:利用逆转录、聚合酶链反应( P C R)等技术从 H F R S恢复期患者外周血淋巴细胞( P B L)中扩增出人 Ig G抗体重链 Fd 基因及κ轻链基因,将之克隆入噬菌体载体p Com b3,构建人源性 Fab 噬菌体抗体基因库. 并利用噬菌体表面呈现技术,对此抗体库进行淘筛、富集. 结果:①成功地构建了人源性 Fab 抗体基因库,库容量为 3×106 . ②从噬菌体抗体库中淘筛得到了能表达人源性抗 H F R S病毒 Fab 抗体的重组克隆. 结论:利用噬菌体抗体库技术,可以获得能表达人源性抗 H F R S病毒抗体的重组克隆.

【Abstract】 AIM: To construct a human Fab phage antibody library and to obtain some recombinant clones which can express Fab antibodies against HFRS virus. METHODS: The total cell RNA extracted from peripheral lymphocytes of HFRS patient was transcribed reversely into cDNA. By amplifying the heavy chain Fd genes and light chain genes of human antibodies from cDNA using semi nested PCR and cloning them into pComb3 vector, a human library of Fab antibodies was constructed, and then it was panned by phage display technology. RESULTS:①A human library of Fab antibodies was constructed, its size is 3×10 6 members. ②Six clones which can express Fab antibodies against HFRS virus were obtained by panning. CONCLUSION: Some recombinant clones which express Fab antibodies against HFRS virus can be obtained by phage display technology.

【基金】 国家自然科学基金
  • 【文献出处】 第四军医大学学报 ,JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY , 编辑部邮箱 ,1999年07期
  • 【分类号】R392.11
  • 【被引频次】5
  • 【下载频次】60
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