| 高致病性禽流感病毒H5N1中和抗体的单链抗体构建与活性鉴定 |
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| 引用本文： | 方钟,罗文新,王明桥,陈瑛炜,张军,陈鸿霖,管轶,夏宁邵.高致病性禽流感病毒H5N1中和抗体的单链抗体构建与活性鉴定[J].生物工程学报,2007,23(2):292-296. |
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| 作者姓名： | 方钟 罗文新 王明桥 陈瑛炜 张军 陈鸿霖 管轶 夏宁邵 |
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| 作者单位： | 1. 厦门大学,国家传染病诊断试剂与疫苗工程技术研究中心,细胞生物学与肿瘤细胞工程教育部重点实验室生命科学学院,厦门,361005
2. 汕头大学医学院,汕港联合流感中心,汕头,515031;香港大学微生物系,新发传染病国家重点实验室,香港 |
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| 基金项目： | 国家禽流感防治专项基金;国家自然科学基金;福建省科技厅科研项目;福建省科研项目;教育部跨世纪优秀人才培养计划;福建省厦门市科技计划 |
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| 摘 要： | 在本实验室研制出的多株针对H5N1血凝素的鼠单抗中,10F7对34株H5N1病毒株都有血凝抑制和中和活性,具有特异性高、反应性强、识别谱广的特点。通过基因工程构建10F7单链抗体(scFv)表达重组质粒,在大肠杆菌中表达并纯化scFv,经血凝抑制实验及中和实验检测其活性。结果在针对3株病毒的血凝抑制实验中,10F7scFv蛋白对其中2株H5N1病毒均显示出结合活性,而对H9毒株没有反应。在针对7株H5N1病毒的中和实验中,10F7scFv对5株病毒具有较好的中和能力。H5N1广谱中和抗体10F7的单链抗体构建,为进一步研制针对H5N1禽流感病毒的治疗性抗体奠定了基础。
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| 关 键 词： | 高致病毒性禽流感病毒 单链抗体 血疑抑制 中和实验 |
| 文章编号： | 1000-3061(2007)02-0292-05 |
| 修稿时间： | 09 11 2006 12:00AM |
Preparation and Identification of a Single-chain Antibody Fragment Against High Pathogenic H5N1 Avian Influenza Virus |
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| FANG Zhong,LUO Wen-Xin,WANG Ming-Qiao,CHEN Ying-Wei,ZHANG Jun,CHEN Hong-Lin,GUAN Yi and XIA Ning-Shao.Preparation and Identification of a Single-chain Antibody Fragment Against High Pathogenic H5N1 Avian Influenza Virus[J].Chinese Journal of Biotechnology,2007,23(2):292-296. |
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| Authors: | FANG Zhong LUO Wen-Xin WANG Ming-Qiao CHEN Ying-Wei ZHANG Jun CHEN Hong-Lin GUAN Yi and XIA Ning-Shao |
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| Institution: | National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen 361005, C;National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen 361005, C;National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen 361005, C;National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen 361005, C;National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen 361005, C;Joint Influenza Research Center (SUMC & HKU), Medical College of Shantou University, Shantou 515031, China;State Key Laboratory of Emerging Infectious Diseases, Department of Microbiology, The University of Hong Kong, Pokfulam, Hong Kong;Joint Influenza Research Center (SUMC & HKU), Medical College of Shantou University, Shantou 515031, China;State Key Laboratory of Emerging Infectious Diseases, Department of Microbiology, The University of Hong Kong, Pokfulam, Hong Kong;National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen 361005, C |
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| Abstract: | Previously,an mAb 10F7 was developed against H5N1 hemagglutinin,whic h was highly specific to 34 different H5N1 strains and showed good neutralizing activity.In the present study,the single-chain fragment of the antibody was c l oned into a prokaryotic vector and then expressed in E.coli.The activity o f the scFv was tested in hemagglution inhibition and neutralization experiment.Two H5N1 virus strains were inhibited to bind erythrocyte cells by the scFv whil e the H9 virus was not.Also,five H5N1 virus strains were neutralized during in fecting MDCK cells.These results showed an approachable method for developing t herapeutic antibody to H5N1 virus. |
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| Keywords: | high pathogenic avian influenza virus single-chain fragment hemagglution inhibition neutralization test |
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