用于新药筛选的新型T4噬菌体纳米颗粒展示平台开发

项目名称： 用于新药筛选的新型T4噬菌体纳米颗粒展示平台开发

项目编号： No.31300652

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 高嵩

作者单位： 淮海工学院

项目金额： 22万元

中文摘要： 在现代新药研发体系中，药物筛选越来越依赖于直接检测药物靶点蛋白与小分子化合物间的亲和性。现有的生物物理学方法对实验样品的制备要求较高，操作复杂，在药物高通量筛选方面的应用受到限制。因此，建立一个简便高效的检测靶点蛋白-小分子化合物亲和性的技术手段具有重要意义。T4噬菌体衣壳是一个良好的蛋白展示纳米颗粒，可高拷贝数展示天然折叠的较大蛋白分子。本项目将运用生化和分子生物学技术手段，建立一套新型的体内同步表达展示法，开发一个T4噬菌体纳米颗粒展示平台，高效快捷地展示活性靶点蛋白。利用这一平台，我们将结合质谱法开发一套便捷方法，检测所展示的靶点蛋白与小分子化合物之间的亲和性；并进一步利用此纳米颗粒展示平台，筛选针对数个热门靶点蛋白的配体化合物，用于下游新药开发。

中文关键词： T4噬菌体；噬菌体展示；毒性蛋白；固定化酶；

英文摘要： Nowadays in drug discovery the drug screening step is more and more dependent on biophysical techniques that can directly evaluate the physical interaction (affinity) between drug target proteins and small molecule compounds. The available biophysical techniques have limitations on their application in drug high-throughput screening because they demand high quality sample preparation and bear complicated procedures. Thus, it is essential to establish a simple and highly efficient technique to measure the affinity between target proteins and small compounds. In this proposal we will develop a phage T4 nano particle platform, based on which the above mentioned affinity measuring technique will be established. Previous studies have indicated that the phage T4 capsid is an excellent nano particle platform for protein display. High molecular weight proteins can be displayed with natural conformations in high copy numbers. By using biochemical and molecular biology methods we will invent a novel in vivo parallel expression method to rapidly display active target proteins on phage T4 nano particles with high efficiency. With this nano particle platform we will also build a simple procedure to measure the affinity between the displayed target proteins and small compounds using mass spectrometry. This nano particle displ

英文关键词： bacteriophage T4；phage display；toxic protein；immobilized enzyme；