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.
出处
《军事医学科学院院刊》
CSCD
北大核心
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)
