Microfluidics-enabled directed affinity reagent engineering for fast, sensitive diagnostics

项目概要/摘要 体液中特定蛋白质的近患者检测和定量可以实现医疗诊断。 专业人员提供及时有效的护理，改善结果。蛋白质生物标志物 检测时，在测定灵敏度和得到结果的时间之间存在折衷。亲和试剂是关键 微流控设备的组件，使蛋白质分析在护理点。目前， 基于平衡结合常数选择亲和试剂，平衡结合常数是最重要的。 在数小时的时间尺度上预测测定性能。该项目将开发 允许亲和试剂工程化的转化动力学筛选方法和装置 可以在几秒到几分钟的时间尺度上提供灵敏的检测， 能够频繁、方便地监测细胞因子组，以改善类风湿性关节炎的护理， 关节炎患者该项目利用了麻省理工学院教师的集体跨学科专业知识 来自化学工程系、电气工程系和计算机系 科学该团队在诊断用蛋白质工程（赛克斯）方面的协同专业知识， 用于即时诊断测试的微流体和BioMEMS的设计、制造和应用 （Voldman）将共同致力于开发新的动力筛选设备， 设计上级亲和试剂，并将新能力转化为有效的现场 炎性细胞因子的测定。具体目标1是创造新的微流体装置， 用于选择快速结合亲和试剂。具体目标2是建立和验证 新的文库筛选过程由设备独特地实现。具体目标3使用快速- 结合亲和试剂，以创建接近患者的细胞因子测定， 灵敏度，无需实验室，可以进行更频繁的测试， 类风湿关节炎的治疗方法

PROJECT SUMMARY/ABSTRACT Near-patient detection and quantification of specific proteins in bodily fluids can enable medical professionals to offer timely and effective care that improves outcomes. In protein biomarker detection, a tradeoff exists between assay sensitivity and time to result. Affinity reagents are key components of microfluidic devices that enable protein analysis at the point of care. At present, affinity reagents are selected on the basis of equilibrium binding constants, which are most predictive of assay performance on hours-long timescales. The proposed project will develop transformative kinetic screening methods and devices that allow engineering of affinity reagents that can offer sensitive detection on the timescales of seconds to minutes, and use the reagents to enable frequent, convenient monitoring of cytokine panels to improve care for rheumatoid arthritis patients. This project leverages the collective cross-disciplinary expertise of faculty at MIT from the Departments of Chemical Engineering and Electrical Engineering and Computer Science. The team's synergistic expertise in protein engineering for diagnostics (Sikes) and design, fabrication and application of microfluidics and BioMEMS for point-of-care diagnostic tests (Voldman) will be collectively focused on developing new kinetic screening devices for engineering superior affinity reagents, and translating the new capability into an impactful on-site assay for inflammatory cytokines. Specific Aim 1 is to create novel microfluidic devices that can be used to select for fast-binding affinity reagents. Specific Aim 2 is to establish and validate new library screening processes uniquely enabled by the devices. Specific Aim 3 uses fast- binding affinity reagents to create a near-patient cytokine assay that maintains the required sensitivity without requiring a laboratory, which can enable more frequent testing and more individualized care for rheumatoid arthritis.