Nanoporous Gold: Extractive Substrate for High-Speed Ultrasensitive Bioassays

纳米多孔金：用于高速超灵敏生物测定的提取底物

基本信息

批准号：
7764726
负责人：
Marc D Porter
金额：
$ 18.32万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-15 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7764726
关键词：
Accounting Acids Address Adhesions Adsorption Aerosols Affinity Alloys Aluminum Oxide Analytical Chemistry Antibodies Antigens Applications Grants Area Arizona Arthropods Back Benchmarking Binding Biocide Biological Biological Assay Biomedical Engineering Bioterrorism Breathing Caliber Calicivirus Capsid Categories Centers for Disease Control and Prevention (U.S.)Chemical Engineering Chemistry Child Clinical Communicable Diseases Complication Coupling Culicidae Data Detection Development Diagnostic Diffusion Digit structure Dimensions Disasters Disease Disease Marker Disease Outbreaks Drops Effectiveness Elderly Electrolytes Emerging Communicable Diseases Engineering Environmental Health Epidemic Esters Ethylene Glycols Event Excision Face Feline Calicivirus Felis catus Figs - dietary Film Flavivirus Fluorescence Food Friction Future Goat Gold Guidelines Health Heating Hepatitis A Hepatitis C virus Hour Housing Human Human Resources Immersion Investigative Technique Immunoassay Individual Infection Ingestion International Investigation Label Laboratories Latex Particles Length Life Ligaments Ligature Link Measurement Measures Membrane Methodology Methods Military Personnel Modeling Modification Monitor Monoclonal Antibodies Morbidity - disease rate Norovirus Oligonucleotides Optics Pathway interactions Performance Phase Phosphate Buffer Planets Polyethylenes Polymers Porcine Parvovirus Porosity Preparation Prevention Procedures Process Proteins Protocols documentation Public Health Quality Control Radial Reaction Reading Regulation Reporting Reproducibility Research Research Design Research Project Grants Rest Rewards Risk Role Rotation Route S-1 Antimetabolite agent Safety Saline Sampling Schools Scientist Senior Scientist Serum Sewage Siloxanes Simulate Solid Solutions Source Specific qualifier value Speed Staging Structure Sulfhydryl Compounds Surface System Tars Techniques Teleconferences Testing Thick Ticks Time Transistors Translating United States Environmental Protection Agency United States National Institutes of Health Universities Upper Respiratory Infections Urbanization Utah Viral Viral Gastroenteritis Viral Proteins Virion Virus Viscosity Water Water Supply West Nile virus Work Yellow Fever analog antigen antibody binding base cross reactivity design drinking water ethylene glycol expectation experience fiberglass fluid flow food surveillance improved innovation interest kinematics light scattering meetings member meter milliliter monolayer mortality nanoparticle nanoscale nanowire pathogen pressure prevent programs public health relevance quality assurance research study response self assembly sensor small molecule stem tool transmission process vector water treatment

项目摘要

DESCRIPTION (provided by applicant): Viruses claimed a significant component of the nearly 15 million lives lost to infectious diseases in 2002. Viral transmission can occur through several routes, including contact with infected individuals, ingestion of contaminated food and water, or contact with a vector like mosquitoes and ticks. These pathways, which may only need to transmit a few tens to hundreds of viruses to trigger an infection, will only become more prevalent as the globalization and urbanization of our planet accelerates. Thus, the ability to detect these and many other pathogens and disease markers rapidly and at very low levels stands as an extremely challenging proposition central to pubic health monitoring, food/water safety, and bioterrorism. While recent breakthroughs have led to the capability to detect viruses and other nanometric targets (e.g., proteins) at single and double digit levels after capture on a solid phase, the time required for sample/label incubation remains a bottleneck for transitioning to the surveillance/monitoring arena. This proposal seeks to redefine assay speed by exploring the potential of nanoporous gold (NPG) to function as a flow-through capture substrate for the efficient extraction of viruses and other comparably-sized pathogens and disease markers (e.g., antibodies), while at the same time accounting for other considerations needed for effective performance. The basis for this strategy rests with predicted improvements in the mass transfer rates, and thus the binding rates, for both the capture and labeling steps in heterogeneous assays that may be realized by flow through a nanoporous material. Models project potential increases in binding rates of more than two orders of magnitude with respect to the most effective of the known approaches. Two groups of experiments are therefore planned to assess this possibility using gold nanoparticle-based surface enhanced Raman scattering (SERS) measurements. In the first group of experiments, NPG membranes of varied pore size will be fabricated, derivatized, and tested as flow through extraction phases for the model virus feline calicivirus, FCV. FCV, which has ~30-nm diameter and is an effective norovirus simulant, will enable an in-depth, systematic assessment of extraction with respect to pore size. These studies will also test the effect of flow rate on capture and label efficiency, and collectively will provide a set of predictive rules for performance optimization in other potential applications. In addition, experiments will be conducted to minimize the impact of nonspecific adsorption by use of blocking agents, as well as potential complications from membrane clogging through the incorporation of sample prefilters. In the second group of experiments, these guidelines will be applied to assays for the detection of FCV in several matrices, including whole goat serum, tap water and groundwater. PUBLIC HEALTH RELEVANCE: This grant proposal seeks to redefine the speed of heterogeneous immunoassays by exploring the potential of nanoporous gold (NPG) membranes to function as flow through capture substrates for the rapid, efficient and selective concentration of nanometrically-sized pathogens (e.g., viruses and proteins). The basis for this strategy rests with: (1) the predicted improvements in the mass transfer rates, and thus the binding rates, for both the capture and labeling steps that may be realized by flow through a nanoporous material; and (2) the high sensitivity of a readout technique that uses modified gold nanoparticles and surface enhanced Raman scattering (SERS). To carry out the above tasks, we have assembled a team of scientists and engineers from the University of Utah Departments of Chemistry, Chemical Engineering, and Bioengineering, and from the Arizona State University School of Materials.

描述（由申请人提供）： 2002 年，病毒夺走了近 1500 万人因传染病而丧生的生命。病毒传播可以通过多种途径发生，包括与感染者接触、摄入受污染的食物和水，或与受感染的人接触。蚊子和蜱虫等媒介。这些途径可能只需要传播数十到数百种病毒即可引发感染，随着地球全球化和城市化的加速，这些途径只会变得更加普遍。因此，快速且低水平地检测这些和许多其他病原体和疾病标志物的能力对于公共卫生监测、食品/水安全和生物恐怖主义来说是一个极具挑战性的命题。虽然最近的突破已经使得能够在固相捕获后以个位数和两位数水平检测病毒和其他纳米目标（例如蛋白质），但样品/标签孵育所需的时间仍然是过渡到监测/监控舞台。该提案旨在通过探索纳米多孔金（NPG）作为流通捕获底物的潜力来重新定义测定速度，以有效提取病毒和其他同等大小的病原体和疾病标记物（例如抗体），同时在同时考虑有效绩效所需的其他考虑因素。该策略的基础在于预测的传质速率的提高，从而提高了结合率，用于异质测定中的捕获和标记步骤，这可以通过流经纳米多孔材料来实现。模型预测，与最有效的已知方法相比，结合率可能增加两个数量级以上。因此，计划进行两组实验，利用基于金纳米粒子的表面增强拉曼散射（SERS）测量来评估这种可能性。在第一组实验中，将制造、衍生化不同孔径的 NPG 膜，并在模型病毒猫杯状病毒 (FCV) 的提取阶段进行测试。 FCV 直径约为 30 nm，是一种有效的诺如病毒模拟物，将能够对孔径的提取进行深入、系统的评估。这些研究还将测试流速对捕获和标记效率的影响，并共同为其他潜在应用中的性能优化提供一组预测规则。此外，还将进行实验，以最大限度地减少使用封闭剂的非特异性吸附的影响，以及通过加入样品预过滤器而导致膜堵塞的潜在并发症。在第二组实验中，这些指南将应用于多种基质中 FCV 的检测，包括全山羊血清、自来水和地下水。公共健康相关性：该拨款提案旨在通过探索纳米多孔金 (NPG) 膜作为流经捕获基质的潜力，以快速、有效和选择性地浓缩纳米级病原体（例如病毒），从而重新定义异质免疫测定的速度。和蛋白质）。该策略的基础在于：（1）对于可以通过流经纳米多孔材料来实现的捕获和标记步骤，预计传质率以及结合率的提高； (2) 使用改良金纳米粒子和表面增强拉曼散射 (SERS) 的读出技术具有高灵敏度。为了完成上述任务，我们组建了一支由来自犹他大学化学系、化学工程系和生物工程系以及亚利桑那州立大学材料学院的科学家和工程师组成的团队。