基于微流控梯度液滴阵列技术的定量高通量筛选—基础研究和生化应用

项目名称： 基于微流控梯度液滴阵列技术的定量高通量筛选—基础研究和生化应用

项目编号： No.21475117

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 数理科学和化学

项目作者： 祝莹

作者单位： 浙江大学

项目金额： 80万元

中文摘要： 定量高通量筛选方法通过在初筛阶段测定样品在多浓度条件下的生物活性，直接得到活性化合物的结构-功能关系，从而有效的消除了常规高通量筛选技术中普遍存在的假阳性和假阴性问题，具有重要的应用前景。然而，定量高通量筛选方法一直受到液体操作繁琐、筛选通量低、试样消耗大等缺点的困扰，导致其难以普及。本项目以研究组的流动注射梯度技术和顺序操作液滴阵列技术为基础，发展多种流动和顺序注射梯度-微流控液滴阵列技术，建立具有高通用型的全自动、超微量的定量高通量筛选方法。结合计算机流体理论模拟和实验方法，建立适用于液滴内浓度梯度预测的理论模型，并发展多种高灵敏度、高动态范围的液滴内样品浓度检测和梯度校正方法。在此基础上，将该方法应用于分子水平的酶抑制剂筛选、细胞水平的化合物毒性测定、以及蛋白质结晶条件筛选等生化筛选领域，为我国的新药研发、毒理学、结构生物学等生物医学领域提供有力的研究工具，促进相关学科的发展。

中文关键词： 微流控芯片；顺序注射梯度；液滴阵列；定量高通量筛选；生化分析

英文摘要： In quantitative high throughput screening (qHTS), each compound is tested at different concentrations against specific biological target to directly obtain the structure-activity relationship of identified compound in primary HTS stage. Thus, qHTS can effectively eliminate the false positive and false negative results that commonly existed in conventional HTS systems. Despite its great potentials, however, qHTS is suffered from complicated liquid handling procedures, low screening throughput, and high sample/reagent consumption. In this proposal, we dedicate to develop a novel sequential injection gradient-microfluidic droplet array (SIGDA) technique to achieve highly versatile quantitative high throughput screening with complete automation and ultra-micro amount sample/reagent consumption. We will combine computational fluid dynamics and experimental methods to build theoretical models for predicting the concentration gradient in droplet array. High sensitive, high dynamic range detection and gradient calibration methods will also be developed to measure the actual concentration in droplets. Based on these fundamental studies, we will apply the SIGDA method in various biological screening systems including enzyme inhibition screening, cell-based toxicity testing, and protein crystallization screening. We envision the SIGDA method can provide a powerful and transformative research tools for drug discovery, toxicology, structural biology, and relative biomedical fields.

英文关键词： Microfluidic chip;Sequential injection gradient;Droplet array;Quantitative high-throughput screening;Bioanalysis