Phase-Changing Sacrificial Layer Microfluidics for Enhanced Protein Analysis

不断开发新的工具和技术，以提高灵敏度、选择性、速度和 生物分析的吞吐量，同时降低每份分析的成本，将是临床诊断的关键， 医学研究，以及生命科学的其他学科。微机械加工方法有可能 能够构建低成本但复杂的微流控系统，用于复杂蛋白质的分析 混合物。在这里，我们建议发展一种相变牺牲层(PCSL)微制造 方法和聚合物表面改性方法，以创建廉价的微流控器件，该微流体器件可以 改进甲胎蛋白(AFP)的检测和定量，甲胎蛋白是一种癌症和胎儿健康的标志 血液和尿液。这项提案的目标有四个方面。首先，我们将推广PCSL溶剂 键合制备技术和原子转移自由基聚合表面改性方法 用于蛋白质分析的聚合物微流控系统系列。其次，我们计划对PCSL溶剂进行评估 用于制造便于样品集成的复杂、多层微流控阵列的键合方法 处理步骤。第三，我们将开发这些制造技术，以使 用于增强检测的免疫亲和纯化系统和蛋白质样品预浓缩器 微芯片毛细管电泳法。第四，我们将利用前三个目标中的工具来制定 从生物中提取和预浓缩甲胎蛋白等低丰度蛋白质的微型设备 然后，我们将评估这些用于测定血液和尿液中甲胎蛋白水平的微系统。 重要的是，虽然本提案中的研究针对的是开发微型化的检测方法 法新社，这些相同的制造和分析程序可以简单地推广到 允许对其他丰度较低的癌症和疾病生物标志物进行快速量化。因此，PCSL 技术和聚合物表面衍生化方法应提供广泛适用的微芯片 用于对人类蛋白质或其他生物分子进行电泳分析和定量的平台。

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.