抗血小板单克隆抗体片段的抗栓效应研究  

Studies on the antithrombotic effects of anti-platelet mAb SZ-21 F(ab′)_2 fragment

作　　者：邵波静[1] 赵益明[1] 沈飞[1] 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血栓室,江苏苏州215006

出　　处：《细胞与分子免疫学杂志》2003年第3期269-271,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目 (No .39830 1 80 )

摘　　要：目的 :研究抗血小板膜糖蛋白IIIa(GPIIIa)单克隆抗体 (mAb)SZ 2 1F(ab′) 2 片段的抗血栓效应。方法 :用木瓜蛋白酶在偏酸条件下 ( pH5 .5 )消化 2 0h ,经DEAE 5 2NaCl梯度洗脱消化产物 ,以得到F(ab′) 2 片段。收集产物经SDS PAGE鉴定 ,并通过ELISA和血小板聚集抑制试验 ,观察所获F(ab′) 2 片段免疫原活性和抗栓活性。结果 :该法制备F(ab′) 2 片段的得率为 6 0 .32 % ;酶免法测定其效价为 0 .8× 10 -9mol/L ;体外血小板聚集抑制半数有效剂量 (ED5 0 )为 3.395mg/L。F(ab′) 2 片段与血小板的结合性及对血小板聚集功能的抑制效果均高于完整的IgG。 结论 :利用木瓜蛋白酶制备的抗血小板mAbSZ 2 1的F(ab′) 2 片段 ,产率高且其抗栓活性与免疫原活性均优于完整的mAbIgG ,为mAbSZ 2AIM: To research the biological effect of the F(ab′) 2 of anti platelet GPIIIa mAb SZ 21. METHODS: The F(ab′) 2 fragment was obtained with papain digestion for 20 hours at 4℃, under acid condition(pH 5.5). The products were purified through DEAE 52 ion exchange column and the first eluted peak components were verified as F(ab′) 2 by SDS PAGE. The immunoactivity and its antithrombotic effects of fragment were detected by ELISA and platelet aggregation inhibition test, respectively. RESULTS: The recovery rate of the fragment was 60.32%, its immunoactivity was 0.8 ×10 9 mol/L by ELISA. The 50% effective dose (ED 50 ) in the platelet aggregate inhibition test was 3.395 mg/L. CONCLUSION: The F(ab′) 2 fragment was superior to intact IgG in binding to platelets and inhibiting platelet aggregation. Thus the F(ab′) 2 fragment of mAb SZ 21 opens up broad prospects for its application to the clinical antithrombotic therapy.

关 键 词：血小板 单克隆抗体 片段 抗栓效应 

分 类 号：R332[医药卫生—人体生理学] R392.11[医药卫生—基础医学]