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.

持续开发新的工具和技术，以提高灵敏度，选择性，速度和生物分析中的吞吐量，同时降低每个测定成本，在临床诊断中至关重要，医学研究以及生命科学中的其他学科。微加工方法有可能启用低成本但复杂的微流体系统的构建，以分析复杂蛋白混合物。本文中，我们建议开发一个改变相位的牺牲层（PCSL）微结构方法和聚合物表面修饰方法可以创建廉价的微流体设备改善α-毒素蛋白（AFP）的检测和定量，一种癌症和胎儿健康标记，血液和尿液。该提案的目标是四倍。首先，我们将概括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用于蛋白质分析的聚合物微流体系统的范围。第二，我们计划评估PCSL溶剂制作复杂的多层微流体阵列的键合方法，以促进样品的整合处理步骤。第三，我们将开发这些制造技术，以实现免疫亲和力纯化系统和蛋白质样品预浓缩剂用于增强检测微芯片毛细管电泳。第四，我们将利用前三个目标的工具来制作提取和预浓缩较低含量蛋白的微型发行条件，例如来自生物学的AFP 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，虽然该提案中的研究是针对开发小型化测定法的法新社，这些相同的制造和分析程序可以简单地推广到允许快速定量其他低估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提供广泛适用的微芯片用于人类中蛋白质或其他生物分子的电泳分析和定量的平台。