人源化抗人纤维蛋白单链抗体-链亲和素融合蛋白的表达和功能

Expression and function of humanized anti-human fibrin single chain antibody-core-streptavidin fusion protein

目的 探索人源化抗人纤维蛋白单链抗体 链亲和素融合蛋白的表达和功能。方法 构建人源化抗人纤维蛋白单链抗体 (ScFv 8E5 )基因与链亲和素基因的重组表达载体 ,制备融合蛋白 ,ELISA观察其稳定性、与人及不同动物纤维蛋白的反应。结果 人源化融合蛋白可特异性识别人、大鼠、豚鼠纤维蛋白 ,同时有生物素的结合位点 ,且稳定性好 ,可耐受至少两周 4℃保存 ,数次反复冻融或一定时间的 37℃孵育。结论 此人源化融合蛋白适用于大鼠及豚鼠血栓病模型 ,是作临床前研究的有用材料。

Objective To express and characterize humanized anti human fibrin single chain antibody core streptavidin fusion protein. Methods The humanized anti human fibrin single chain antibody core streptavidin fusion protein was constructed by inserting a sequence encoding core streptavidin at the 3′ end of the humanized ScFv 8E5 structural gene. ELISA was used in analysis of its antigen binding ability. Results The fusion protein was stable, capable of binding to human, rat and guinea pig fibrin, but not to their fibrinogen and bovine albumin. Conclusion Humanized anti fibrin singe chain antibody core streptavidin fusion protein could be useful in future study on thrombus imaging and also thrombolytic agents targeting.