抗纤维蛋白单克隆抗体的制备和鉴定  

PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES SPECIFIC FOR FIBRIN AND ITS FRAGMENTS

作　　者：奚晓东[1] 赵益明[1] 王庆才[1] 李佩霞[1] 白霞[1] 周泉生[1] 阮长耿[1] 

机构地区：[1]江苏省血液研究所苏州医学院血栓与止血研究室,苏州215007

出　　处：《中国实验血液学杂志》1994年第2期181-188,共8页Journal of Experimental Hematology

基　　金：江苏省科委资助课题(H9202)

摘　　要：用XL-FN颗粒、D-D、FN单体等作为免疫原获得一组小鼠单克隆抗体。除了3个与FG有交叉反应,其余均只与XL-FN和/或其片段反应。其中SZ-58、SZ-60及SZ-63与血浆凝块的结合反应不受血浆或全血影响。在免疫印迹电泳中,SZ-63主要识别D-D,而SZ-58不识别任何区带。SZ-58、SZ-60可抑制血浆凝固,并对ADP诱导的血小板聚集有程度不同的抑制作用。SZ-60可与固相化的FG结合而与溶解态的FG无结合反应。结果提示,SZ-58、SZ-60、SZ-63对XL-FN反应的特异性较高,因此可应用于体内血栓显像而SZ-63同时又可应用于D-D检测。Fibrinogen is a large dimer molecule composed of three nonidentical polypeptide chains. In the presence of thrombin, it converts to fibrin by cleavage of fibrinopeptide A and fibrinopeptide B, Mr 1 800 each, and forms crossliked fibrin by γ-γ cross links mediated by factor XⅢ. This conversion results in retention of 98% of the primary structure of the precursor molecule. Therefore, it is difficult to obtain a specific antibody to discriminate fibrinogen and fibrin. In this study, we tried to produce the monoclonal antibody specific for the conformational epitope expressed only on fibrin but not on fibrinogen. Nine monoclonal antibodies have been raised from spleen cells of BALB / c mice immunized with particles of crosslinked fibrin, D-Dimer and fibrin monomer. Six of them specifically reacted with crosslinked fibrin and / or its fragments. Three others cross-reacted with immobilized fibrinogen. Binding of SZ-58, SZ-60 and SZ-63 could scarcely be affected by the presence of plasma or whole blood. In western blot , SZ-63 predominantly recognized D-Dimer while no band could be identified by SZ-58. On the other hand , SZ-58, SZ-60 and SZ-65 inhibited plasma coagulation caused by thrombin and SZ-58, SZ-60 and SZ-64 inhibited ADP-induced platelet aggregation to different extents. We also found that the epitope for SZ-60 was only expressed on the fibrinogen immobilized on some surfaces but not on soluble fibrinogen. Our data suggest that SZ-58, SZ-60 and SZ-63 possess relatively high specificity to crosslinked fibrin and hence they have potential in thrombus imaging in vivo. In addition, SZ-63 could be applied in D-Dimer detection and SZ-64 and SZ-65 be helpful to immunoassay for fibrinogen.

关 键 词：纤维蛋白 纤维蛋白原 单克隆抗体 

分 类 号：R392[医药卫生—免疫学]