Microfluidic MALDI-MS Device for High-Throughput Proteomics & Biomarker Discovery

描述（由申请人提供）：癌症是美国死亡的主要原因，每年诊断出超过1，000，000例新病例。由于大规模人群筛查所需的高灵敏度和特异性，目前在临床环境中常规使用的单一蛋白质生物标志物很少。当务之急是开发新的技术，能够快速检测与早期诊断、预后、分期和治疗反应相关的大量生物标志物。 这项研究的长期目标是将新兴的微流体技术与最先进的质谱（MS）检测相结合，以提高我们分析生物系统中分子结构和功能的能力。该应用充分利用了微流体架构的独特能力，可实现过程集成、多路复用、快速和高通量处理微量样品，以及MS检测的能力，可提供明确检测痕量组分所需的灵敏度、特异性和分辨率。 具体目标1：开发一种紧凑、低成本和一次性的微流控分析平台，具有基质辅助激光解吸电离（MALDI）-MS/MS检测功能，用于高通量蛋白质组学，从而能够研究蛋白质共表达模式和生物标志物发现。微流控装置将进行平行液相色谱（LC）分离，并将集成一个新的微芯片-MS接口，使灵敏的MALDI-MS/MS检测直接从芯片。 具体目标二：展示微流控MALDI-MS/MS平台检测细胞提取物中多种癌症生物标志物的有效性。将分析MCF 7乳腺癌细胞系的细胞组分中参与驱动癌症发生和发展的基本细胞过程（细胞增殖、细胞周期调控、DNA修复、细胞凋亡和侵袭/转移）的靶蛋白。

DESCRIPTION (provided by applicant): Cancer is a leading cause of death in the United States with over 1,000,000 new cases being diagnosed every year. As a result of the high sensitivities and specificities that are required to justify large-scale population screening, only very few single protein biomarkers are routinely used today in the clinical setting. It is of critical priority to develop novel technologies that will enable the rapid detection of a plethora of biomarkers relevant to early diagnosis, prognosis, staging and treatment response. The long-term objective of this research is to combine the emerging technology of microfluidics with state-of-the-art mass spectrometry (MS) detection to enhance our capacity for analyzing molecular structure and function in biological systems. This application capitalizes on the distinguishing capabilities of microfluidic architectures that enable process integration, multiplexing, fast and high-throughput processing of minute amounts of sample, and the power of MS detection that provides the sensitivity, specificity and resolving power necessary for unambiguous detection of trace level components. Specific Aim 1: Develop a compact, low-cost and disposable microfluidic analysis platform with matrix assisted laser desorption ionization (MALDI)-MS/MS detection for high-throughput proteomics that will enable the study of protein co-expression patterns and biomarker discovery. The microfluidic device will carry out parallel liquid chromatography (LC) separations and will integrate a novel microchip-MS interface to enable sensitive MALDI-MS/MS detection directly from the chip. Specific Aim 2: Demonstrate the effectiveness of the microfluidic MALDI-MS/MS platform for the detection of multiple cancer biomarkers in cellular extracts. Cellular fractions from the MCF7 breast cancer cell line will be analyzed for target proteins that are involved in essential cellular processes driving cancer on-set and development (cell proliferation, cell cycle regulation, DNA repair, apoptosis and invasion/metastasis).