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

支持微流体的定向亲和试剂工程可实现快速、灵敏的诊断

• 批准号: 10527811
• 负责人: HADLEY D SIKES
• 金额: $ 21.66万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10527811
• 关键词: Affinity; Autoimmune Diseases; Binding; Biological; Biological Assay; Biological Products; Biology; Caring; Cell Separation; Chemical Engineering; Chemicals; Chronic; Detection; Development; Devices; Diagnostic; Diagnostic tests; Dissociation; Electrical Engineering; Elements; Engineering; Enzyme-Linked Immunosorbent Assay; Equilibrium; Faculty; Future; Generations; Goals; Gold; Health; Healthcare; Home; Hour; Inflammatory; Interleukin-1 beta; Interleukin-17; Interleukin-6; Investigation; Kinetics; Laboratories; Libraries; Liquid substance; Medical; Medicine; Methods; Microfluidic Microchips; Microfluidics; Modeling; Monitor; Monoclonal Antibodies; Patients; Penetration; Performance; Process; Protein Analysis; Protein Engineering; Proteins; Rapid diagnostics; Reagent; Receptor Signaling; Reproducibility; Rheumatoid Arthritis; Scheme; Science; Site; Specificity; Speed; Surface; System; TNF gene; Testing; Therapeutic; Time; Tissues; Translating; Yeasts; base; biomarker panel; chronic autoimmune disease; clinically relevant; combinatorial; computer science; cytokine; design; design-build-test; effective intervention; improved; improved outcome; mathematical model; monitoring device; novel; pathogen; personalized care; point of care; point-of-care diagnostics; predictive test; protein biomarkers; screening; tumor

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.

项目概要/摘要 靠近患者的体液中特定蛋白质的检测和定量可以使医疗 专业人员提供及时有效的护理，以改善结果。在蛋白质生物标志物中 检测时，检测灵敏度和得出结果的时间之间存在权衡。亲和试剂是关键 能够在护理点进行蛋白质分析的微流体装置的组件。现在， 亲和试剂的选择基于平衡结合常数，这是最 预测数小时的时间尺度上的测定性能。拟议项目将开发 允许亲和试剂工程化的变革动力学筛选方法和装置 可以在秒到分钟的时间尺度上提供灵敏的检测，并使用试剂 能够频繁、方便地监测细胞因子组，以改善类风湿病的护理 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 (Sikes) 和 用于现场诊断测试的微流体和 BioMEMS 的设计、制造和应用 （沃尔德曼）将共同致力于开发新的动力学筛选设备 设计卓越的亲和试剂，并将新功能转化为有影响力的现场 炎症细胞因子测定。具体目标 1 是创建新型微流体装置 用于选择快速结合的亲和试剂。具体目标 2 是建立并验证 这些设备独特地启用了新的文库筛选过程。具体目标 3 使用快速 结合亲和试剂创建近患者细胞因子测定，维持所需的 无需实验室即可实现更频繁的测试和更多的灵敏度 类风湿性关节炎的个体化护理。