抗人BlyS嵌合抗体的研制及功能鉴定  

Development and functional identification of an anti-rhBlyS chimeric antibody

作　　者：林周[1] 程建贞[1] 谷欣[1] 冯健男[1] 黎燕[1] 沈倍奋[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《军事医学科学院院刊》2007年第3期232-234,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：国家"863"课题重点项目资助(No.2006AA020503)

摘　　要：目的:在具有中和功能的母本抗体基础上,进行人源化改造,获得具有相同功能的嵌合抗体。方法:本研究以一株具有中和活性的抗人BlyS鼠单抗(B20)为母本,将抗人BlyS鼠单抗B20的轻、重链可变区基因分别插入到含有人κ链和IgG1重链恒定区基因的真核表达载体中,构建了人-鼠嵌合抗体表达载体,转染真核细胞,通过ELISA、RT-PCR、免疫印迹、体外中和实验分别对嵌合抗体(CB20)进行检测。结果:RT-PCR结果显示,转染后细胞系有人-鼠嵌合抗体mRNA的转录;ELISA、免疫印迹实验鉴定表明,该嵌合抗体与人BlyS特异性结合;体外中和实验表明,此抗体能中和BlyS对B淋巴细胞促增殖效应。结论:成功地构建人-鼠嵌合抗体CB20。Objective：To develop an chimerical antibody which has the same neutral function as the parent antibody Mab B20 has. Methods： A cAb CB20 was reconstructed from mAb MB20 which consisted of murine antigen binding region （from MB20） and human constant region （from human IgG1 ）. The VH-express/CB20 and VK-express/CB20 were trans- fected into the 293T cell line for eukaryotic expression. Then the supernant of transfected 293T cell was identified. Results：The RT-PCR analysis showed that the transfected 293T cells had the gene transcription of human-mouse antibody. ELISA and Western blot assay showed that the CB20 had recognition specificity to BlyS. The in vitro neutralization assay proved that CB20 could neutralize the proliferation-promoting effect of BlyS on B lymphocytes. Conclusion： CB20 humanmurine chimeric antibody is successfully constructed.

关 键 词：BLYS 人-鼠嵌合抗体CB20 真核表达 中和抗体 

分 类 号：R392.11[医药卫生—免疫学]