Phase-Changing Sacrificial Layer Microfluidics for Enhanced Protein Analysis

描述（由申请人提供）：不断开发新的工具和技术，提高生物分析的灵敏度、选择性、速度和吞吐量，同时降低每次分析的成本，将对临床诊断、医学研究和生命科学的其他学科至关重要。微加工方法有潜力使低成本的建设，但复杂的微流体系统分析复杂的蛋白质混合物。在此，我们提出了一种相变牺牲层（PCSL）微加工方法和聚合物表面改性方法，以创造廉价的微流体装置，可以提高血液和尿液中甲胎蛋白（AFP）的检测和定量，甲胎蛋白是癌症和胎儿健康的标志。这项建议的目标有四个方面。首先，我们将把PCSL溶剂键合制造技术和原子转移自由基聚合表面修饰方法推广到一系列用于蛋白质分析的聚合物微流控系统中。其次，我们计划评估PCSL溶剂键合方法用于制作复杂的多层微流体阵列，以促进样品处理步骤的集成。第三，我们将开发这些制造技术，使免疫亲和纯化系统和蛋白质样品预浓缩器的结合能够用于增强微芯片毛细管电泳的检测。第四，我们将利用前三个目标的工具制造从生物混合物中提取和预浓缩低丰度蛋白质（如AFP）的微型设备；然后我们将评估这些微系统，以确定血液和尿液中的AFP水平。重要的是，虽然本提案中的研究是针对AFP的小型化检测的发展，但这些相同的制造和分析程序可以以一种直接的方式推广，以允许其他低丰度癌症和疾病生物标志物的快速定量。因此，PCSL技术和聚合物表面衍生化方法应该为人类蛋白质或其他生物分子的电泳分析和定量提供一个广泛适用的微芯片平台。

DESCRIPTION (provided by applicant): The continued development of new tools and techniques that improve sensitivity, selectivity, speed and throughput in biological analysis, while reducing the cost per assay, will be critical in clinical diagnosis, medical research, and other disciplines in the life sciences. Micromachining methods have the potential to enable the construction of low-cost, yet sophisticated microfluidic systems for the analysis of complex protein mixtures. Herein, we propose to develop a phase-changing sacrificial layer (PCSL) microfabrication approach and polymer surface modification methods to create inexpensive microfluidic devices that can improve the detection and quantification of alpha-fetoprotein (AFP), a cancer and fetal wellness marker, in blood and urine. The objectives of this proposal are four-fold. First, we will generalize the PCSL solvent bonding fabrication technique and atom-transfer radical polymerization surface modification methods to a range of polymer microfluidic systems for protein analysis. Second, we plan to evaluate the PCSL solvent bonding approach for making complex, multilayer microfluidic arrays that facilitate the integration of sample processing steps. Third, we will develop these fabrication techniques to enable the incorporation of immunoaffinity purification systems and protein sample preconcentrators for use in enhancing detection in microchip capillary electrophoresis. Fourth, we will utilize the tools from the first three objectives to make microdevices that extract and preconcentrate lower-abundance proteins such as AFP from biological mixtures; we will then evaluate these microsystems for the determination of AFP levels in blood and urine. Importantly, while the studies in this proposal are directed toward the development of miniaturized assays for AFP, these same fabrication and analysis procedures can be generalized in a straightforward manner to allow the rapid quantification of other lower-abundance cancer and disease biomarkers. Thus, PCSL techniques and polymer surface derivatization methods should provide a broadly applicable microchip platform for the electrophoretic analysis and quantification of proteins or other biomolecules in humans.