人源性抗血小板膜糖蛋白Ⅱb/Ⅲa Fab抗体的研制及其对血小板聚集功能的影响  

Generation of human Fab antibody against platelet membrane glycoprotein Ⅱb/Ⅲa and its effect on platelet aggregation

作　　者：董宁征[1] 崔宇杰[1,2] 阮长耿[1] 

机构地区：[1]苏州大学附属第一医院、江苏省血液研究所、卫生部血栓与止血重点实验室,江苏苏州215006 [2]天津医科大学医学检验系,天津300020

出　　处：《细胞与分子免疫学杂志》2009年第1期65-67,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省基础研究计划(自然科学基金)资助课题(BK2005036)

摘　　要：目的:构建人源性抗血小板膜糖蛋白Ⅱb/Ⅲa(GPI-Ib/IIIa)Fab噬菌体抗体库,筛选抗GPⅡb/Ⅲa特异性的噬菌体抗体,并对其功能进行研究。方法:取血小板膜糖蛋白抗体(抗GPIIb/IIIa)阳性的特发性血小板减少性紫癜(ITP)患者脾细胞,采用噬菌体抗体库技术构建人源性抗GPⅡb/Ⅲa Fab噬菌体抗体库,以表达GPⅡb/Ⅲa的CHO123细胞筛选抗体库,并以ELISA法检测噬菌体抗体;用Western blot对抗体进行鉴定,并测定其与血小板抗原的结合;观察筛选到Fab抗体对血小板聚集的影响。结果:筛选出2株能够与血小板膜GPⅡb/Ⅲa特异性结合的Fab抗体,其序列与人免疫球蛋白轻、重链可变区序列具有高度同源性,表达纯化的Fab抗体能抑制血小板聚集。结论:成功地从人源性抗血小板膜糖蛋白Ⅱb/Ⅲa Fab抗体库筛选出特异性识别GPⅡb/Ⅲa,具有抑制血小板聚集作用的人源性抗体。AIM： To generation of human Fab fragment against GP GPIIb/IIIa from phage display library and to study its effect on platelet aggregation. METHODS： An antibody phage display library was constructed from spleen cells from a donor with idiopathic thrombocytopenic purpura （ITP） whose anti-GPIIb/IIIa antibody was positive. High-affinity human mAbs was selected by panning against GPIIb/ IIIa expressed on CHO123 cells. The binding specificity of phage antibody to GPIIb/IIIa was detected by ELISA and Westem blot. And the effect of antibody on platelet aggregation was analyzed. RESULTS： After three rounds of panning with CHO123, phage antibodies against GPIIb/IIIa were enriched specifically. 2 positive phage antibodies with high specific for GP IIb/IIIa were verified, and the purified antibody can inhibited ADP induced platelet aggregation. CONCLUSION： Human antibodies against GPIIb/IIIa are obtained from phage display library.

关 键 词：血小板膜糖蛋白IIB/IIIA 噬菌体展示 FAB 血小板聚集 

分 类 号：R392.11[医药卫生—免疫学]