| 抗基孔肯亚病毒人-鼠嵌合抗体真核表达载体的构建和表达 |
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| 引用本文： | 李建民,陈薇,安小平,樊英茹,郝晓萌,李冰,陈万荣,刘树玲,童贻刚.抗基孔肯亚病毒人-鼠嵌合抗体真核表达载体的构建和表达[J].军事医学科学院院刊,2004,28(3):225-228. |
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| 作者姓名： | 李建民 陈薇 安小平 樊英茹 郝晓萌 李冰 陈万荣 刘树玲 童贻刚 |
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| 作者单位： | 军事医学科学院微生物流行病研究所,北京,100071 |
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| 基金项目： | 总后“十五”指令课题 ( 18-D5 0 1d) |
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| 摘 要： | 目的：表达具有中和活性的抗基孔肯亚病毒人．鼠嵌舍抗体。方法：用RT-PCR法克隆具有中和活性的抗基孔肯亚病毒鼠源单克隆抗体CHIK-IF1的轻重链可变区基因，克隆人IgG1恒定区基因组基因，构建了轻重链嵌舍基因和表达质粒pCdhfrlL，pcDNA3、1H，共转染二氢叶酸还原酶缺陷型CHO细胞，用ELISA检测培养上清中嵌舍抗体的表达，MTX加压筛选表达量高的克隆，扩大培养收集上清并纯化，纯化的嵌合抗体进行SDS-PAGE、Western印迹检测和抗原结合活性的检测。结果与结论：在CHO细胞中成功表达了抗基孔肯亚病毒的嵌舍抗体，为制备抗基孔肯亚病的被动免疫制剂奠定了良好的基础。
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| 关 键 词： | 基孔肯亚病毒 免疫球蛋白基因 人-鼠嵌合抗体 真核表达 CHO细胞 |
| 文章编号： | 1000-5501(2004)03-0225-04 |
Expression of human-mouse chimeric antibody directed against Chikungunya virus |
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| LI Jian-Min,CHEN Wei,AN Xiao-Ping,FAN Ying-Ru,HAO Xiao-Meng,LI Bing,CHEN Wan-Rong,LIU Shu-Ling,TONG Yi-Gang.Expression of human-mouse chimeric antibody directed against Chikungunya virus[J].Bulletin of the Academy of Military Medical Sciences,2004,28(3):225-228. |
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| Authors: | LI Jian-Min CHEN Wei AN Xiao-Ping FAN Ying-Ru HAO Xiao-Meng LI Bing CHEN Wan-Rong LIU Shu-Ling TONG Yi-Gang |
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| Abstract: | Objective: To express human-mouse chimeric antibody directed against Chikungunya virus. Methods: An anti-CHIK human-mouse chimeric antibody was expressed by cloning the light and heavy chain variable region gene of the anti-CHIK mouse monoclonal antibody CHIK-IF1 by RT-PCR , and then was used to construct the light and heavy chain expressing plasmids pCdhfr1L and pcDNA3.1H. Results and Conclusion: The human-mouse chimeric antibodies were expressed in CHO cells. The successful expression may provide a passive immunization drug for the Chikungunya disease. |
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| Keywords: | Chikungunya virus immunoglobulin genes human-mouse chimeric antibody eukaryotic expression CHO cells |
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