重组SZ51Hu-scuPA融合蛋白的体外溶栓研究  

Study on thrombolysis of a recombinant SZ51 Hu scuPA in vitro

作　　者：万海英[1] 刘征辉[1] 顾津明 朱明清[2] 李佩霞[1] 阮长耿[2] 

机构地区：[1]苏州医学院附属第一医院江苏省血液研究所血栓与止血研究室,215006 [2]苏州医学院核医学重点实验室

出　　处：《江苏医药》2000年第10期772-774,共3页Jiangsu Medical Journal

基　　金：国家"863"高技术科学发展计划基金!(102090302)

摘　　要：目的　获得大量人源化抗人活化血小板单抗SZ5 1Hu和尿激酶原 (scuPA)融合蛋白SZ5 1Hu scuPA并测定该基因重组抗体导向溶栓剂的导向溶栓性。方法　通过转瓶扩大培养系统获得大量SZ5 1Hu scuPA ;通过抗体竞争结合实验证明融合蛋白的抗体亲和力 ;进而与尿激酶进行比较对不同浓度血小板血浆凝块的溶栓研究。结果　该融合蛋白在培养上清中的表达量为 5mg/ml;其纤溶活性为 390 0 0IU/mg ;抗体亲和力是鼠源性单抗的 6 7% ;体外溶栓效力较uPA提高 4 1～ 8 4倍。结论　该融合蛋白比uPA具有更高、更快的溶栓效率 。Objective To compare the thrombolytic properties in vitro between urokinase(uPA) and the recombinant chimeric plasminogen activator SZ51 Hu scuPA,which was composed of a humanized monoclonal antibody SZ51(SZ51 Hu) specifically against activated human platelet and a single chain urokinase(scuPA).Methods The fusion protein SZ51 Hu scuPA was produced from the supernatant of a stable myeloma cell line grown in spinner flask.Its affinity to activated human platelet was obtained by competitive binding of SZ51 Hu scuPA and murine mAb SZ51;the thrombolytic potency of SZ51 Hu scuPA was compared with uPA by a system composed of an 125 I fibrin labeled human plasma clot.Results The concentration of the fusion protein in the supernatant was 5mg/ml.The binding capacity to activated human platelet of purified fusion protein SZ51Hu scuPA was 67% of the SZ51 M and the fibrinolytic activity was 39,000 IU/mg.In the dissolution of the human plasma clots containing the different concentration of human platelets,the fusion protein SZ51Hu scuPA was 4 1 8 4 folds more potent than uPA.Conclusion The fusion protein SZ51Hu scuPA was more competent to dissolve platelet clots than uPA.

关 键 词：重组SZ51Hu-scuPA 融合蛋白 体外溶栓 

分 类 号：R973.2[医药卫生—药品]