单细胞功能蛋白质组学微流控芯片新方法研究

项目名称： 单细胞功能蛋白质组学微流控芯片新方法研究

项目编号： No.21275060

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 数理科学和化学

项目作者： 刘笔锋

作者单位： 华中科技大学

项目金额： 80万元

中文摘要： 从单个细胞水平探讨生命过程已成为近年来生命分析化学新的挑战和机遇。目前，国际单细胞蛋白质组学主要采用化学细胞术或流式细胞术两条截然不同路线。本项目提出建立一种单细胞功能蛋白质组学新方法，以微流控芯片为平台，采用流式细胞术中蛋白质功能性标记策略，利用单色荧光抗体集对单细胞功能蛋白质组或群进行识别和标记，并进一步采用化学细胞术策略，在微流控芯片上完成细胞的运送、定位、溶胞和蛋白质分子电泳分离与激光诱导荧光检测，从而克服了化学细胞术在蛋白质鉴定上的困扰及峰容量问题，也解决了流式细胞术多色复杂标记的难点及大量原代细胞消耗的困境；采用上述新思路，重点针对中国人群发病率最高的癌症肝癌，研究单个细胞水平癌症信号相关蛋白组的差异表达，并描画出肝癌信号调控网络。本课题对于生命分析化学、蛋白质科学和癌症等基础研究以及个体化医疗等应用领域具有重要意义，并促进分析化学及其相关交叉学科的发展和人才培养。

中文关键词： 单细胞；蛋白质组；微流控芯片；异质性；肿瘤

英文摘要： Investigation on life at single cell level has become the new challenge for bioanalytical chemistry in recent years. Currently, single cell proteomics is manly achieved by two thoroughly different strategies such as chemical cytometry and flow cytometry. In this project, we demonstrate a novel method to realize single cell functional proteomics. In brief, on a microfluidic chip cell proteins are functionally identified and labeled by fluorescently-tagged antibodies (idea learned from flow cytometry strategy). The labeled cells are then transported and selected for subsequent on-line single cell lyses, electrophoretic separation and highly sensitive laser-induced fluorescence detection (idea learned from chemical cytometry strategy). This new method not only overcomes problems in proteins identification and peak capacity of chemical cytometry, but also overcomes problems in the use of multi-color antibodies and high cost on primary cells of flow cytometry. With this novel roadmap, we will focus our interest in liver cancer, a tumor disease of the highest risk in Chinese population, and try to characterize the expressions of cancer-related signal transduction proteome, and map out the network of liver cancer signal regulations as well. This method paves a new avenue to early cancer detection and personalized medic

英文关键词： Single cell；Proteomics；Microfluidic chip；Heterogeinity；Tumor