以高亲和力结合血红蛋白β链与δ链单克隆抗体的研制  

Preparation of high-affinity monclonal antibody against hemoglobin δ globin chain and β globin chain

作　　者：朱平[1] 肖海燕[1] 陈益国[1] 徐梅[1] 富宁[1] 

机构地区：[1]南方医科大学基础医学院免疫学教研室,广东广州510515

出　　处：《细胞与分子免疫学杂志》2010年第6期556-559,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：广州市社会发展计划资助项目(06A1212008)

摘　　要：目的:制备可同时识别血红蛋白A2(HbA2)与血红蛋白A(HbA)的单克隆抗体(mAb),即与血红蛋白的δ链与β链结合,而不与γ链结合。方法:以原核表达的重组Hbδ链免疫小鼠,常规融合制备杂交瘤细胞,以离子交换层析分离的天然HbA2与HbA为抗原同时进行筛选;所获mAb用间接ELISA、基于变性与非变性聚丙烯凝胶电泳的Western blot、等离子表面共振及流式细胞术进行鉴定。结果:所获mAb2C9以高亲和力与纯化的天然HbA2与HbA结合,经等离子表面共振测定KA值分别为2.25×1011、2.16×1010;该mAb不与胎儿血红蛋白(HbF)、血红蛋白α链及重组血红蛋白ζ(zeta)链结合。结论:获得1株可结合HbA2与HbA的mAb2C9,提示其识别血红蛋白δ链与β链的共同表位。该抗体将成为研究与诊断血红蛋白病的有效工具。AIM：To prepare monclonal antibody （mAb） against both hemoglobin A2 （HbA2） and hemglobin A （HbA）, this antiboy can bind to δ globin chain and β globin chain, but not to γ globin chain. METHODS： Mice was immunized with recombinant Hb δ fusion protein, and hybridoma cells were generated by cell fusion techniques followed by screening with natural HbA2 and HbA separated by ion exchange chromatography. The purified monclonal antibody was identified by indirect ELISA, Western blot based on native PAGE and SDS-PAGE, surface plasmon resonance （SPR）, flow cytometry and immunohistochemistry. RESULTS： The monoclonal antibody against both HbA2 and HbA was obtained and designated as 2C9 that shows no binding to fetal hemoglobin （HbF）, α globin chain and recombinant ζ globin chain. CONCLUSION： The mAb 2C9 was defined as specificity to hemoglobin δ globin chain and β globin chain, which suggests that mAb 2C9 recognizes a common epitope on bothδglobin chain and β globin chain. This antibody would be expected to be an effective tool in research and clinical practice in hemoglobinopathies.

关 键 词：血红蛋白δ链 Β链 HBA2 HBA 单克隆抗体 

分 类 号：Q813.2[生物学—生物工程]