文档介绍：
利用 BIAcore 3000 建立抗 His 标签抗体捕获
的 HIV 病毒蛋白药物筛选模型#
王娇,马洪涛,胡秦**
5
10
15
20
25
(北京工业大学生命科学与生物工程学院,北京 100124)
摘要:目的:基于表面等离子体共振(Surface plasmon resonance, SPR)技术,建立以艾
滋病病毒整合酶和 Vpu 蛋白为靶点的高通量药物筛选模型。方法:选择 BIAcore 3000 系统
(GE 公司)和葡聚糖修饰的 CM5 芯片,将抗 His 标签抗体(TA02 和 E02)偶联到传感芯片
表面后利用抗体捕获整合酶和 Vpu 蛋白,通过响应值(Ru)大小确定抗体的检测限并验证蛋
白的结合活性。结果:抗 His 标签抗体(TA02 和 E02)均可捕获整合酶蛋白,经抗体捕获的
Vpu 蛋白与 Tetherin 具有良好的结合能力。结论:基于 SPR 技术建立的艾滋病病毒蛋白靶
向药物筛选模型具有好的应用前景。
关键词:表面等离子体共振技术;抗 His 抗体;HIV 整合酶;Vpu
中图分类号:Q67;Q74
Build a screening model of HIV proteins targeted drugs
Using Surface Plasmon Resonance
WANG Jiao, MA Hongtao, HU Qin
(Life Science and Bioengineering College,Beijing University of Technology,Beijing 100124)
Abstract: A surface plasmon resonance(SPR) biosensor-based assay was developed for
high-throughput screening of HIV integrase and Vpu protein inhibitors. Method: SPR experiments
were performed on a BIAcoreTM 3000(GE Healthcare). His tagged HIV integrase and Vpu
proteins were captured on anti-his antibodies immobilized to the CM5 chips. The binding
efficiency pared between 2 antibodies and biological activity was verified using Tetherin
protein. Results: two antibodies were both able to efficiently capture integrase protein and
captured Vpu retained Tehterin-binding : the SPR-based screening assay has
shown great potential for anti-HIV drug discovery in fut