噬菌体展示蛋白质芯片的无标记检测  

Label-free detection of phage-displayed protein chips

作　　者：刘芳芳[1] 罗昭锋[2] 定翔[1] 朱圣庚[3] 余兴龙[1] 

机构地区：[1]清华大学精密仪器与机械学系,精密测试技术与仪器国家重点实验室,北京100084 [2]中国科学技术大学生命科学实验中心,合肥230027 [3]北京大学生命科学学院,北京100871

出　　处：《清华大学学报（自然科学版）》2008年第11期1935-1939,共5页Journal of Tsinghua University(Science and Technology)

基　　金：国家自然科学基金资助项目(30727001);高等学校博士学科点专项科研基金资助项目(20050003056)

摘　　要：为获得丰富的特异识别分子和实现无标记、高灵敏度检测,利用噬菌体展示技术,选择在pIII蛋白上展示12肽的噬菌体M13作探针,采用羧基-氨基共价耦联法制备噬菌体展示蛋白质芯片。在此基础上,与表面等离子体共振传感技术结合,使用Biacore3000仪器,检测噬菌体展示蛋白质芯片与特异性抗体的反应,测定反应动力学常数。实验结果表明:当抗体浓度为0.4μmol.L-1时,响应信号可达365±8个单位;不同浓度的抗体与芯片反应的数据较好地与S型曲线吻合。该方法可望用于噬菌体展示蛋白质芯片的检测。Surface plasmon resonance and the phage display technique were combined to enable use of a variety of specific reagents for label-flee, high sensitivity detection in protein chips. Phage M13 which displays 12 peptides on its coated protein plII was selected as the probe to prepare the phage-displayed protein chip using the amine coupling method. The specific interactions between the chip and the antibodies were detected using Biacore3000, with the binding kinetic parameters determined. Tests show that an antibody concentration of 0. 4μmol .L-1 gives response units of 365 ：k 8, and that the relationship between the binding responses and the various antibody concentrations is well fit by the sigrnoid curve. The method can be used to optimize the detecting conditions for phage-displayed protein chips.

关 键 词：表面等离子体共振 噬菌体M13展示系统 噬菌体展示蛋白质芯片 特异性抗体 动力学常数 

分 类 号：R197.39[医药卫生—卫生事业管理]