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  

Expression of human-mouse chimeric antibody directed against Chikungunya virus

作　　者：李建民[1] 陈薇[1] 安小平[1] 樊英茹[1] 郝晓萌[1] 李冰[1] 陈万荣[1] 刘树玲[1] 童贻刚[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《军事医学科学院院刊》2004年第3期225-228,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：总后"十五"指令课题 ( 18-D5 0 1d)

摘　　要：目的 :表达具有中和活性的抗基孔肯亚病毒人 鼠嵌合抗体。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 ,构建了轻重链嵌合基因和表达质粒pCdhfr1L ,pcDNA3.1H ,共转染二氢叶酸还原酶缺陷型CHO细胞 ,用ELISA检测培养上清中嵌合抗体的表达 ,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合抗体 ,为制备抗基孔肯亚病的被动免疫制剂奠定了良好的基础。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.

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 

分 类 号：R392.11[医药卫生—免疫学]