抗人CD25嵌合抗体基因的构建及其瞬时表达研究  

Study on construction and transient expression of human-mouse chimeric antibody gene against human CD25

作　　者：胡迪超[1] 张爱华[1] 潘勇兵[1] 詹珊珊[1] 杨晓明[1] 

机构地区：[1]武汉生物制品研究所,武汉430060

出　　处：《中国免疫学杂志》2011年第7期648-653,共6页Chinese Journal of Immunology

摘　　要：目的:构建抗人CD25嵌合抗体基因并在哺乳动物细胞中进行瞬时表达和初步鉴定。方法:采用RLM-RACE法克隆WuTac抗体可变区和信号肽序列,并利用基因拼接法构建嵌合抗体基因。用脂质体法瞬时转染三种哺乳动物细胞,并使用ELISA、FCM、WB、Dot blot和免疫荧光法进行检测。结果:成功克隆WuTac抗体可变区和信号肽序列,并构建了抗人CD25嵌合抗体表达质粒。瞬时转染结果表明所表达的嵌合抗体保留了亲本抗体WuTac的抗原结合力。结论:成功构建了抗人CD25嵌合抗体基因,为其进一步研究打下基础。Objective：To construct chimeric antibody gene against human CD25 antigen,and preliminarily identify the expressed products produced from transiently transfected mammalian cells in order to facilitate the further study of stable expression.Methods：The RLM-RACE was employed to clone variable region genes and leader sequences,and the Overlap PCR method was used to construct the chimeric antibody gene.After transiently transfected in three mammalian cells with liposome method,the expressed products were determined by ELISA,FCM,WB,Dot blot and immunofluorescence assay.Results：The variable region genes and leader sequences were successfully amplified,and the eukayotic expression plasmids were constructed.The results from transient transfection indicate the expressed products retain the antigen binding capacity of parental antibody WuTac.Conclusion：The successfully constructed chimeric antibody gene against human CD25 lays a sound foundation for further study.

关 键 词：CD25 RLM—RACE 基因拼接 嵌合抗体 瞬时表达 

分 类 号：R392.11[医药卫生—免疫学]