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: 7194205
• 负责人: Adam Thomas Woolley
• 金额: $ 26.22万
• 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 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7194205
• 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; Facility Construction Funding Category; 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; Object Attachment; Performance; Polyethylene Terephthalates; Polymers; Polystyrenes; Procedures; Process; Protein Analysis; Proteins; Range; Sampling; Solutions; Solvents; Speed; Standards of Weights and Measures; Surface; System; Systems Analysis; Techniques; Testing; Urine; alpha-Fetoproteins; base; clinical Diagnosis; clinically relevant; cost; desire; fetal; improved; microchip; microsystems; miniaturize; phase change; polycarbonate; polymerization; size; tool

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）：不断开发新的工具和技术，提高生物分析的灵敏度、选择性、速度和通量，同时降低每次测定的成本，这对于临床诊断、医学研究和其他学科至关重要。生命科学。微加工方法有可能构建低成本但复杂的微流体系统，用于分析复杂的蛋白质混合物。在此，我们建议开发一种相变牺牲层（PCSL）微加工方法和聚合物表面修饰方法，以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，我们将 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体系统。其次，我们计划评估 PCSL 溶剂键合方法，用于制造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，以实现免疫亲和纯化系统和蛋白质样品预浓缩器的结合，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中的工具来制造微型设备，从生物混合物中提取和预浓缩低丰度蛋白质，例如 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，虽然本提案中的研究旨在开发 AFP 的小型化测定，但这些相同的制造和分析程序可以以直接的方式推广，以允许快速量化其他低丰度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该为人类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。