Two-Dimensional Electrophoresis Device for Biomarker Detection

描述（由申请人提供）：拟研究的目标是开发用于药物发现和医学诊断的蛋白质组学技术。具体来说，我们的目标是开发一种微流体驱动的二维电泳设备，它有可能取代传统的二维凝胶电泳（2DGE）。除了流行的蛋白质组学技术，如质谱（MS）和蛋白质微阵列，2DGE仍然是蛋白质组学研究中最有效的方法之一。2DGE的主要优势是能够将样品与对照进行比较，以鉴定疾病的生物标志物，而其主要局限性是可重复性差和耗时的过程。

DESCRIPTION (provided by applicant): The goal of the proposed research is to develop proteomics technology for drug discovery and medical diagnostics. Specifically we aim to develop a microfluidics-enabled, two-dimensional electrophoresis device that has a potential to replace conventional two-dimensional gel electrophoresis (2DGE). In addition to popular proteomics techniques such as mass spectrometry (MS) and protein microarrays, 2DGE remains one of the most potent methods in proteomics studies. The key advantage of 2DGE is its ability to compare samples with controls for identification of biomarkers of diseases while its major limitations are poor reproducibility and time-consuming processes. To address the limitations, we will develop a 2D electrophoresis device consisting of one channel for the first dimension (isoelectric focusing) and a number of orthogonal channels for the second dimension (polyacrylamide gel electrophoresis). An array of microfluidic pseudovalves will be created for introducing different separation media; a fluidic network allows the transfer of proteins from the first to the second dimension. Faster electrophoresis and higher separation resolution are expected due to higher separation voltage, in an analogy to DNA sequencing that has shifted from slab gel electrophoresis to the capillary array format. In addition, the device will provide enhanced reproducibility of 2D maps due to elimination of gelwarping and improved accuracy in protein identification resulting from the elimination of the diffusion and interaction among proteins in adjacent gel lanes. The specific aims include: (1) designing and fabricating the 2D electrophoresis device consisting of an array of microvalves and the fluidic network; (2) demonstrating the device by protein analysis; and (3) validating the device by analyzing the biomarkers associated with traumatic brain injury (TBI). The significance of the research lies in the fact that the 2D electrophoresis device has the potential to replace 2DGE, which is widely used in biological laboratories. Secondly, if the platform is successfully implemented, it will likely become a rapid, disposable, and quantifiable diagnostic tool to help physicians determine the seriousness of TBI and to guide implementation of appropriate medical treatment. In addition, the platform will likely be useful in searching for the association between biomarkers and a variety of other diseases, and in screening/diagnosing a particular disease using the established biomarkers. Public Health Relevance Statement: The proposed research is to develop proteomics technology for drug discovery and medical diagnostics. Specifically we will develop a microfluidics-enabled electrophoresis device that has a potential to replace classical two-dimensional slab gel electrophoresis. The device will be used for detecting the biomarkers associated with traumatic brain injury.