EAGER: Elastomeric Capture Microparticles for High Sensitivity Biodection

智能优点：在基于生物特异性相互作用的检测之前进行分离可以显著提高信噪比，从而提高生物传感测量的特异度、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特别有吸引力，因为可以最大限度地减少对流和扩散对配体结合的限制，同时消除了通过膜或柱的高压泵送的需要。此外，微粒子的受体部位占有率可以通过常规的实验室方法进行分析，如流式细胞术或平板读数。这项提议展示了声学分离微粒的概念验证，并说明了如何将这一现象发展为增强分子、病毒和细胞分析物生物传感的连续分离战略。从复杂的样品环境(例如血液或其他含有细胞的悬浮液)中分离目标分析物是通过在弹性微粒上显示生物分子受体来实现的，弹性微粒在施加在微流体流上的声波驻波中表现出负对比。声辐射使弹性微粒从细胞等不可压缩微粒中分离出来。从复杂样品组分中去除目标分析物可以进行精确的化学分析，例如通过荧光、电泳法或质谱学手段。该项目将执行三项不同的任务，包括合成稳定的弹性微粒子，对这些微粒子进行生物特异性相互作用的修饰，以及验证分子和细胞分析物的分离和检测方法。因此，这些任务的完成将为弹性粒子与声学分离一起用于高性能生物分析操作提供基本的基础。更广泛的影响：这项初步工作将为新型弹性微粒在多种生物传感方式中的使用奠定基础，包括生物分子传感、稀有细胞检测和细胞分离。开发的原理、材料和方法将适用于包括食品安全、环境监测、过程控制和国防在内的许多领域的生物检测。这项工作还将建立材料合成和生物功能化的方法，使弹性体微米和纳米颗粒能够用于体内成像、传感和靶向药物输送。该项目将资助一名女性生物化学家博士后的工资，她的目标是在生物传感器科学和工程领域获得更多经验。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。这个项目的人员将通过利用杜克大学现有的人力资源来增加，包括本科生研究人员(例如，由NSF REU计划或杜克·普拉特研究员计划支持)和进行独立研究以获得研究生学分的硕士研究生。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.