抗血小板膜糖蛋白单克隆抗体SZ-2单链抗体的构建、表达及功能研究

被引：8

作者：

戴克胜

祝怀平

阮长耿

机构：

[1] 江苏省核医学生物技术重点实验室

[2] 苏州大学附属第一医院江苏省血液研究所

来源：

中国免疫学杂志 | 2003年 / 04期

关键词：

单链抗体; 单克隆抗体; 血小板;

D O I：

暂无

中图分类号：

R392 [医学免疫学];

学科分类号：

100102 ;

摘要：

目的 :降低鼠源性抗体的免疫原性及分子量 ,为其进一步研究、应用奠定基础。方法 :应用RT PCR技术 ,克隆SZ 2重链、轻链可变基因。应用基因重组技术构建SZ 2单链抗体表达载体pET2 2 2scFv ,导入大肠杆菌BL2 1(DE3)Plys ,诱导表达。流式细胞术、ELISA、Westernblot检测SZ 2scFv与血小板结合能力 ,瑞斯托霉素、凝血酶和ADP诱导血小板聚集试验对表达产物功能进行研究。结果 :克隆基因序列符合小鼠轻、重链可变区基因特征 ;pET2 2 2scFv表达质粒拼接正确 ;表达产物以包涵体形式为主 ,表达量占菌体总蛋白的 2 5 % ;纯化、复性后证明表达产物具有与血小板结合的活性 ,并可抑制瑞斯托霉素、凝血酶诱导的血小板聚集。结论 :成功表达了SZ 2单链抗体 ,该小分子抗体具有与血小板结合的活性 ,并可抑制瑞斯托霉素、凝血酶诱导的血小板聚集。

引用

页码：243 / 247+252 +252

页数：6

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