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| 抗H5N1病毒嵌合IgA抗体基因的构建及其在CHO细胞中的表达 | |
| 引用本文： | 张宝中,张昕,陈万荣,刘大斌,王盛,安小平,冉多良,赵光宇,周育森,童贻刚.抗H5N1病毒嵌合IgA抗体基因的构建及其在CHO细胞中的表达[J].生物工程学报,2009,25(5):714-719. |
| 作者姓名： | 张宝中 张昕 陈万荣 刘大斌 王盛 安小平 冉多良 赵光宇 周育森 童贻刚 |
| 作者单位： | 1. 军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京,100071;新疆农业大学动物医学学院,乌鲁木齐,830052 2. 军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京,100071 3. 新疆农业大学动物医学学院,乌鲁木齐,830052 |
| 基金项目： | 国家自然科学基金项目(No.30872223);;国家科技支撑计划项目(No.2006BAD06A15)资助~~ |
| 摘 要： | 为了表达具有中和活性的抗禽流感H5N1病毒人-鼠嵌合IgA抗体,采用RT-PCR法克隆具有中和活性的抗禽流感H5N1-HA鼠源单克隆抗体的轻重链可变区基因及相应的信号肽编码序列,分别与人免疫球蛋白IgA2重链恒定区、Kappa恒定区基因拼接,构建表达质粒pEF-IGHA9和pEF-IGK9,共转染二氢叶酸还原酶缺陷型CHO(CHO-dhfr-)细胞,用ELISA检测培养上清中嵌合IgA抗体的表达,对纯化的嵌合抗体进行SDS-PAGE、Western blotting印迹分析。结果成功地在CHO细胞中表达了抗禽流感H5N1病毒人-鼠嵌合IgA抗体,为制备抗H5N1重组分泌型IgA预防性抗体制剂奠定了良好的基础。 |
| 关 键 词： | H5N1病毒 免疫球蛋白基因 嵌合IgA抗体 CHO细胞 |
| 收稿时间： | 2008/12/10 0:00:00 |
Construction of anti-H5N1 virus chimeric IgA antibody gene and its expression in CHO cells |
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| Baozhong Zhang,Xin Zhang,Wanrong Chen,Dabin Liu,Sheng Wang,Xiaoping An,Duoliang Ran,Guangyu Zhao,Yusen Zhou and Yigang Tong.Construction of anti-H5N1 virus chimeric IgA antibody gene and its expression in CHO cells[J].Chinese Journal of Biotechnology,2009,25(5):714-719. | |
| Authors: | Baozhong Zhang Xin Zhang Wanrong Chen Dabin Liu Sheng Wang Xiaoping An Duoliang Ran Guangyu Zhao Yusen Zhou and Yigang Tong |
| Institution: | State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China; College of Veterinary Science, Xinjiang Agricultural University, Urmuqi 830052, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;College of Veterinary Science, Xinjiang Agricultural University, Urmuqi 830052, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China |
| Abstract: | To express human-mouse chimeric IgA antibody directed against H5N1 virus, an anti-H5N1 chimeric IgA antibody gene was constructed by joining the light and heavy chain variable region genes and the corresponding signal peptide coding sequences of the anti-H5N1 mouse monoclonal antibody H5N1-HA with the coding sequences of the constant region of the human IgA2 heavy chain and Kappa chain respectively.Then the full-length chimeric light and heavy chain expressing plasmids pEF-IGHA9 and pEF-IGK9 were constructe... |
| Keywords: | H5N1 virus immunoglobulin gene chimeric IgA antibodies CHO cells |
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