Phase-Changing Sacrificial Layer Microfluidics for Enhanced Protein Analysis

用于增强蛋白质分析的相变牺牲层微流体

• 批准号: 7079019
• 负责人: Adam Thomas Woolley
• 金额: $ 27万
• 依托单位: BRIGHAM YOUNG UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7079019
• 关键词: blood; microfluidics; phase change; polymers; proteins; solvents; urine

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.

描述（由申请人提供）：持续开发新的工具和技术，这些工具和技术在生物分析中提高灵敏度，选择性，速度和吞吐量，同时降低每个测定的成本，对于临床诊断，医学研究和生活科学中的其他学科至关重要。微加工方法有可能实现低成本但复杂的微流体系统来分析复杂蛋白质混合物。本文中，我们建议开发一种改变相位的牺牲层（PCSL）微生物方法和聚合物表面修饰方法，以创建廉价的微流体设备，以改善α-五蛋白蛋白（AFP）的检测和量化，一种癌症和胎儿健康标记，在血液和尿液中。该提案的目标是四倍。首先，我们将概括PCSL溶剂键合的制造技术和原子 - 转移自由基聚合表面修饰方法，以用于一系列聚合物微流体系统进行蛋白质分析。其次，我们计划评估PCSL溶剂键合方法，以制造复杂的多层微流体阵列，以促进样品处理步骤的整合。第三，我们将开发这些制造技术，以使免疫亲和力纯化系统和蛋白质样品预浓缩剂掺入以增强微芯片毛细管电泳中的检测。第四，我们将利用前三个目标中的工具来制作微论条件，从而提取和预浓缩降低含量较低的蛋白质，例如生物混合物中的AFP。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，尽管该提案中的研究是针对法新社的小型化测定法的开发，但可以直接地将这些相同的制造和分析程序推广，以允许快速量化其他低估的癌症和疾病生物标志物。因此，PCSL技术和聚合物表面衍生化方法应为人类的蛋白质或其他生物分子的电泳分析和定量提供广泛适用的微芯片平台。