抗血小板糖蛋白单克隆抗体的纯化及生物活性鉴定  

Purification of Anti-platelet Glycoprotein Monoclonal Antibody and Characterization of Its Biological Activity

作　　者：杨永昌[1] 王北宁[1] 赖春宁[2] 黎燕[2] 

机构地区：[1]北京军区总医院检验科,北京100700 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《标记免疫分析与临床》2008年第3期156-158,共3页Labeled Immunoassays and Clinical Medicine

摘　　要：原核表达重组质粒p3MH/P140-κFd,在E.coliXLI-Blue菌中获得了成功表达,利用重组蛋白中的6个组氨酸(His)组成的"标签"进行亲和层析,对表达产物进行纯化。SDS-PAGE蛋白电泳和Western blot的检测结果证明,纯化得到的分子量约47kD蛋白是鼠源性的Fab抗体,ELISA检测结果证明,P140Fab抗体与人血小板能特异性结合。体外二磷酸腺苷(ADP)诱导的血小板聚集抑制试验证明,P140Fab抗体对血小板的聚集有抑制作用,并且具有剂量抑制效应,IC50的平均值为16.85mg/L。Recombinant plasmid p3MH/P140κ-Fd was expressed successfully in E. coli XLI- Blue. The expression product with hiss tag was purified by metal affinity resins. The molecular weight of soluble protein was 47kD and identified to be a Fab fragment of mouse anti-human monoclonal antibody by SDS-PAGE and Western blot. The result of ELISA showed that P140 Fab can be specific bound to platelet. The platelet aggragation inhibition test indicated that P140 Fab could inhibit platelet aggragation specific in a dose dependent manner. The mean value of IC50 was 16.85 mg/L.

关 键 词：血小板 单克隆抗体 蛋白纯化 生物活性 

分 类 号：R914[医药卫生—药物化学]