基于二硫化碳改性表面的无细胞表达系统构建蛋白质芯片感应表面的研究

项目名称： 基于二硫化碳改性表面的无细胞表达系统构建蛋白质芯片感应表面的研究

项目编号： No.21305147

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 数理科学和化学

项目作者： 牛宇

作者单位： 中国科学院力学研究所

项目金额： 25万元

中文摘要： 全内反射椭偏成像蛋白质芯片作为一种新型蛋白质相互作用检测分析方法，具有重大应用前景。但是，该蛋白质芯片的感应表面，仍存在着配基活性表现不佳和非特异性吸附的问题，阻碍了该蛋白质芯片进一步的应用。针对现有不足，本项目拟从配基分子表达、芯片表面改性和配基装配方法入手，着重考虑如何提高配基分子活性表现和阻隔非特异性吸附等科学问题，同时关注蛋白质芯片反应界面性质和光学信号传导等因素，耦合利用无细胞表达系统和二硫化碳表面改性技术，尝试在金表面上进行配基分子的表达和原位活性装配，探索构建配基分子数量可控、具有高生物活性表现和能够抑制非特异性吸附的实用化感应表面的方法。在此基础上，针对感应表面的性质，优化椭偏光学器件设置，以期提高该蛋白质芯片的检测灵敏度和动态范围等性能指标，推进其在生物学和临床医学领域的应用。本项目的预期研究结果具有广泛适用性，能够用于以金表面为基底的其它蛋白质相互作用分析手段。

中文关键词： 蛋白质芯片；感应表面；无细胞表达系统；二硫化碳表面改性；

英文摘要： The protein chip based on total internal reflection imaging ellipsometry has been a novel but promising technique for protein-protein interaction analysis. However, several drawbacks in its biosensing interface remain a significant barrier for its development and further application in the field of fundamental biology and clinical laboratory test. In order to overcome these problems, the studies on the expression method and immobilization approach of protein ligand, as well as surface modification strategy for protein chip interface are planned to carry out in this investigation. Focusing on the crucial function of biosensing interface, cell-free protein synthesis system and carbon disulfide surface modification method are combined to construct a practical biosensing interface which is expected to present good bio-activity of protein ligand and the capability to inhibit non-specific adsorption. Thereafter, according to the feature of the biosensing interface and the principle of ellipsometry signal response, the setup of the optical system is optimized to improve its detection sensitivity and dynamic range, prompting its biology and clinic application. The desired achievements of this project can not only be used for the protein chip based on total internal reflection imaging ellipsometry, but also fulfill the n

英文关键词： Protein Chip；Biosensing Interface；Cell-Free Protein Synthesis System；Carbon Disulfide Surface Modification；