抗H5N1病毒嵌合IgA抗体基因的构建及其在CHO细胞中的表达  

Construction of anti-H5N1 virus chimeric IgA antibody gene and its expression in CHO cells

作　　者：张宝中[1,2] 张昕[1] 陈万荣[1] 刘大斌[1] 王盛[1] 安小平[1] 冉多良[2] 赵光宇[1] 周育森[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病原微生物生物安全国家重点实验室,北京100071 [2]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《生物工程学报》2009年第5期714-719,共6页Chinese Journal of Biotechnology

基　　金：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。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-HSN1 mouse monoclonal antibody HSN1-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 constructed and transfected into the CHO/dhfr cells. The chimeric IgA antibody expression was confirmed by ELISA, SDS-PAGE and Western blotting. The successful expression of this anti-H5N1 chimeric IgA may help to provide a stand for developing passive immunological agents for HSN 1 virus infection prophylaxis.

关 键 词：H5N1病毒 免疫球蛋白基因 嵌合IgA抗体 CHO细胞 

分 类 号：R392[医药卫生—免疫学]