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

项目摘要/摘要 近病人对体液中特定蛋白质的检测和量化可以使医疗 专业人员提供及时和有效的护理，以改善结果。在蛋白质生物标志物中 在检测过程中，在分析灵敏度和结果时间之间存在权衡。亲和力试剂是关键 微流控设备的组件，可在护理时进行蛋白质分析。目前， 亲和试剂是根据平衡结合常数来选择的，平衡结合常数是 对长达数小时的时间尺度上的分析性能进行预测。拟议中的项目将开发 允许亲和试剂工程化的转化动力学筛选方法和设备 可以在几秒到几分钟的时间尺度上提供灵敏的检测，并使用试剂 实现对细胞因子面板的频繁、方便的监测，以改善对类风湿的护理 关节炎患者。这个项目利用了麻省理工学院教员的集体跨学科专业知识。 来自化学工程、电气工程和计算机系 科学。该团队在用于诊断的蛋白质工程(SIKES)和 点诊断式微流控和生物微机械的设计、制造和应用 (Voldman)将集体专注于开发新的动态筛选设备 设计卓越的亲和力试剂，并将新功能转化为现场有效的 炎性细胞因子的检测。具体目标1是创造新型微流控设备，该设备可以 用于选择快速结合的亲和试剂。具体目标2是建立和验证 由这些设备独一无二地启用的新库筛选流程。《特定目标3》使用FAST- 结合亲和试剂以创建近患者细胞因子检测，以保持所需的 无需实验室即可实现更频繁的检测和更多 类风湿性关节炎的个体化护理。

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.