EAGER: Elastomeric Capture Microparticles for High Sensitivity Biodection

智力优势：在基于生物特异性相互作用的检测之前的分离可以显著增加信噪比水平，并且因此可以增强生物感测测量中的特异性、灵敏度和置信度。基于悬浮液中微粒的生物特异性相互作用的分离是特别有吸引力的，因为对配体结合的对流和扩散限制可以最小化，而同时避免了对通过膜或柱的高压泵送的需要。此外，微粒的受体位点占有率可以通过常用的实验室方法如流式细胞术或平板阅读进行常规分析。该提案展示了微粒声学分离的概念验证，并说明了这种现象如何发展成为一种连续的分离策略，用于增强分子，病毒和细胞分析物的生物传感。从复杂样品环境（例如，血液或其它含细胞的悬浮液）是通过在弹性微粒上显示生物分子受体来实现的，所述弹性微粒在施加于微流体流上的声学驻波中显示负对比度。声辐射导致弹性体微粒与不可压缩的颗粒如细胞分离。从复杂样品组分中除去目标分析物允许精确的化学分析，通过荧光、电泳或质谱方法。该项目将执行三个不同的任务，包括合成稳定的弹性微粒，修饰这些微粒的生物特异性相互作用，以及验证分子和细胞分析物的分离和检测方法。因此，这些任务的实现将提供基本的基础，通过该基础，弹性体颗粒可以与声学分离结合用于高性能的生物分析操作。更广泛的影响：这项初步工作将为在许多生物传感模式中使用新型弹性体微粒奠定基础，包括生物分子传感，稀有细胞检测和细胞分离。所开发的原理、材料和方法将适用于许多背景下的生物检测，包括食品安全、环境监测、过程控制和国防。这项工作还将建立材料合成和生物功能化的方法，这将使弹性体微米和纳米粒子的体内成像，传感和靶向药物输送的检查。该项目将支持一名女性生物化学博士后研究员的工资，其目标是在生物传感器科学和工程领域获得进一步的经验。通过这个项目，她还将在生物医学工程的几个方面获得宝贵的经验，包括生物材料界面工程，微流体和生物分析仪器。该项目的人员将通过利用杜克现有的人力资源来增加，包括本科研究人员（例如，由NSF REU计划或杜克普拉特研究员计划支持）和硕士水平的研究生谁进行研究生学分的独立学习研究。这些机制中的每一个都将用于加强该项目的教育和研究培训影响。

Intellectual Merit: Separation prior to biospecific interaction based detection can significantly increase signal-to-noise level and thus can enhance specificity, sensitivity, and confidence in biosensing measurements. Separations based on biospecific interaction of microparticles in suspension are particularly attractive because convective and diffusional limitations to ligand binding can be minimized, while at the same time obviating the need for high pressure pumping though membranes or columns. Furthermore, receptor site occupancy of microparticles can be routinely analyzed by common laboratory methods such as flow cytometry or plate reading. This proposal demonstrates proof-of-concept for acoustic separation of microparticles and illustrate how this phenomenon can be developed into a continuous separation strategy for enhanced biosensing of molecular, viral and cellular analytes. Separation of a target analyte from a complex sample milieu (e.g., blood or other cell-containing suspension) is accomplished by displaying biomolecular receptors on elastomeric microparticles that exhibit negative contrast in acoustic standing waves imposed on microfluidic streams. Acoustic radiation results in the separation of elastomeric microparticles from incompressible particles such as cells. Removal of the target analyte from the complex sample components allows precise chemical analysis, e.g., by fluorometric, electrophoretic or mass spectrometric means. This project will execute three distinct tasks including synthesis of stable elastomeric miroparticles, modification of these microparticles for biospecific interaction, and validation of separation and detection methods for molecular and cellular analytes. Achievement of these tasks will thus provide the fundamental basis by which elastomeric particles can be used in conjunction with acoustic separations for high performance bioanalytical manipulations. Broader Impacts: This preliminary work will form the basis for the use of new types of elastomeric microparticles in a number of biosensing modalities, including biomolecular sensing, rare cell detection and cell isolation. The principles, materials and methods developed will be applicable to biodetection in a number of contexts including food safety, environmental monitoring, process control and national defense. This work will also establish methods for materials synthesis and biofunctionalization that will enable the examination of elastomeric micro- and nanoparticles for in vivo imaging, sensing and targeted drug delivery. This project will support the salary of a female biochemist postdoctoral fellow, whose goal is to gain further experience in the field of biosensor science and engineering. Through this project, she will also gain valuable experience in several aspects of biomedical engineering, including biomaterial interfacial engineering, microfluidics and bioanalytical instrumentation. The personnel on this project will be augmented by taking advantage of human resources available at Duke, including undergraduate researchers (e.g., supported by NSF REU program or Duke Pratt Fellows program) and Masters level graduate students who conduct independent study research for graduate credit. Each of these mechanisms will be used to enhance the educational and research training impact of this project.