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

基本信息

批准号：
8212420
负责人：
Adam Thomas Woolley
金额：
$ 23.31万
依托单位：
BRIGHAM YOUNG UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8212420
关键词：
Affinity Antibodies Benchmarking Biological Biological Assay Biological Markers Biological Sciences Blood Blood specimen Buffers Capillary Electrophoresis Complex Complex Mixtures Detection Development Devices Discipline Disease Drug Formulations Fluorescence Genetic Crossing Over Human Injection of therapeutic agent Ionic Strengths Ions Label Liquid substance Malignant Neoplasms Medical Research Membrane Methodology Methods Microchip Analytical Procedures Microchip Electrophoresis Microfabrication Microfluidic Microchips Microfluidics Modification Performance Polyethylene Terephthalates Polymers Polystyrenes Procedures Process Protein Analysis Proteins Sampling Solutions Solvents Speed Surface System Systems Analysis Techniques Testing Urine alpha-Fetoproteins base clinical Diagnosis clinically relevant cost fetal improved microchip microsystems miniaturize phase change polycarbonate polymerization protein complex tool

项目摘要

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.

不断开发新的工具和技术，提高灵敏度、选择性、速度和生物分析的通量，同时降低每次检测的成本，对于临床诊断至关重要，医学研究和生命科学的其他学科。微加工方法有潜力能够构建低成本但复杂的微流体系统来分析复杂的蛋白质混合物。在此，我们建议开发相变牺牲层（PCSL）微加工方法和聚合物表面改性方法来创建廉价的微流体装置，可以改善甲胎蛋白 (AFP)（一种癌症和胎儿健康标志物）的检测和定量血液和尿液。该提案有四个目标。首先，我们对PCSL溶剂进行概括键合制造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体系统系列。其次，我们计划评估PCSL溶剂用于制造复杂的多层微流体阵列的粘合方法，有利于样品的集成处理步骤。第三，我们将开发这些制造技术，以便能够将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。第四，我们将利用前三个目标中的工具来实现从生物体中提取并预浓缩 AFP 等低丰度蛋白质的微型装置混合物；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旨在开发小型化分析方法法新社，这些相同的制造和分析程序可以以简单的方式推广到允许快速量化其他丰度较低的癌症和疾病生物标志物。因此，PCSL 技术和聚合物表面衍生方法应提供广泛适用的微芯片用于对人体蛋白质或其他生物分子进行电泳分析和定量的平台。