抗CD3/抗CD20双特异双链抗体的生物学活性研究  

Study on the biological activity of anti-CD3/anti-CD20 diabody

作　　者：熊冬生[1] 许元富[1] 杨纯正[1] 彭晖[1] 邵晓枫[1] 刘汉芝[1] 韩俊岭[1] 赖增祖[1] 朱祯平[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所,天津300020

出　　处：《中华微生物学和免疫学杂志》2001年第6期627-631,共5页Chinese Journal of Microbiology and Immunology

基　　金：86 3计划资助项目 ( 10 2 0 9 0 3 0 3)

摘　　要：目的　研究抗CD3 抗CD2 0双特异双链抗体的生物学活性。方法　采用亲和层析法纯化本室构建的抗CD3 抗CD2 0双特异双链抗体可溶性表达产物 ,并用SDS PAGE ,Westernblot和分子排阻层析鉴定纯化产物 ;采用FACS法和玫瑰花环试验测定纯化产物与靶细胞的结合活性 ;采用3 H TdR掺入实验和51 Cr释放试验测定该双特异双链抗体的生物学性质。结果　纯化的抗CD3 抗CD2 0双特异双链抗体具有与Jurkat(CD3+)和Daudi细胞 (CD2 0 +)的结合活性 ,且能同时与Jurkat和Daudi细胞结合形成玫瑰花环 ,并能竞争性封闭亲代鼠源性抗体HIT3a和HI47与Jurkat和Daudi细胞的结合位点 ;该双特异双链抗体具有促有丝分裂原作用和介导激活的T细胞杀伤Daudi细胞的活性。结论　抗CD3 抗CD2 0双特异双链抗体具有与亲代鼠源性抗体HIT3a和HI47相同的性质 ,且能介导激活的T细胞杀伤表达CD2 0抗原的肿瘤细胞 ,是一个有望用于B细胞恶性肿瘤临床治疗的双特异性抗体。Objective To study the biological activity of anti CD3/anti CD20 diabody. Methods The anti CD3/anti CD20 diabody was purified by affinity chromatography and analyzed by both Western blot and size exclusion chromatography. Its antigen binding activity was examined by FACS and rosetting assay; its biological activity was examined by 3H TdR incorporation and 51 Cr release test. Results The anti CD3/anti CD20 diabody bound to Jurkat cells (CD3 +) and Daudi cells(CD20 +) and block the sites where parent antibody bind to. Furthermore, it was found to have effective mitogenic activity and the effect on retargeting peripheral blood lymphocytes to lyse Daudi cells. Conclusion The biological activity of anti CD3/anti CD20 diabody is the same as its parent antibodies. The diabody proved to be a potent agent for retargeting peripheral blood lymphocytes to tumor cells expressing the CD20 antigen and then lyse them. [

关 键 词：CD3 CD20 基因工程抗体 双特异性抗体 B细胞 恶性肿瘤 治疗 

分 类 号：R392[医药卫生—免疫学]