Refreshable Biosensors for Continuous Renal Function Monitoring

项目摘要 所提出的探索性研究的目标是设计和开发新的可刷新生物传感器， 克服与现有生物传感器相关的关键基本挑战，以实现实时，定量， 连续监测复杂体液中的蛋白质生物标志物，包括尿液、血液、间质液、唾液 和汗水。常规技术，如酶联免疫吸附测定（ELISA）、蛋白质印迹和免疫印迹法， 质谱法使得能够发现和研究生物样品中的蛋白质。这些常规 由于生物识别的稳定性差 元素此外，它们依赖于额外的试剂或pH变化来去除结合的分析物以实现可重复使用性。 这些传统方法与用于连续分子生物学的可穿戴和可植入装置不兼容。 监测.因此，迫切需要开发可刷新的生物传感器， 定量、连续监测复杂体液中的蛋白质生物标志物。在这次调查中， 采用肾损伤（阿基）作为疾病模型。各种蛋白质生物标志物，包括 金属蛋白酶-2、胰岛素样生长因子结合蛋白7和中性粒细胞明胶酶相关脂质运载蛋白 已被确定用于阿基的早期诊断/预后。每种生物标志物在特定的时间范围内都有特定的时间过程 伤害的设置。连续监测与生理和病理生理相关的一组阿基生物标志物， 考虑到阿基可以迅速发展，因此在受伤组织中的过程非常重要。但目前的 蛋白质定量技术不能确定在临床环境中何时实际发生损伤，并且 无法提供实时肾功能评估，以及时进行临床干预。长期目标是 通过开发一种完全无线、小型化、多功能的生物传感器， 可以直接连接到导尿管或患者膀胱的内表面。的总体目标 本计画旨在设计与发展一种新颖的可更新生物感测器，并以实验验证其连续性即时性。 监测患者尿液中的阿基蛋白生物标志物。具体目标包括：1.生物识别的设计与实现 元件具有高灵敏度、特异性和稳定性，可用于可更新的生物传感器。根据可行性研究， 通过聚合物的分子印迹获得的生物识别元件对靶蛋白表现出高亲和力 生物标志物，并且在极端条件下具有化学、热和机械稳定性，这对于 所提出的可刷新生物传感器。2.设计、开发和验证刺激器， 表面声能来更新生物识别元件。主要假设是局部和病灶 所递送的高表面声能可以解吸特异性结合的靶蛋白并刷新 生物传感器的生物识别位点。这种新的能力对于监测肾功能是非常有价值的 存在AKI风险或患有阿基的危重患者，包括脓毒症、休克、大手术和创伤患者。

Project Summary The goal of the proposed exploratory research is to design and develop novel refreshable biosensors that overcome key fundamental challenges associated with existing biosensors to achieve real-time, quantitative, continuous monitoring of protein biomarkers in complex body fluids, including urine, blood, interstitial fluid, saliva and sweat. Conventional techniques such as enzyme-linked immunosorbent assay (ELISA), Western Blot, and mass spectrometry have enabled discovery and study of proteins in biological samples. These conventional bioassays are typically limited to one-time or short time use due to the poor stability of biological recognition elements. Furthermore, they rely on additional reagents or pH change to remove bound analytes for reusability. These traditional processes are not compatible with wearable and implantable devices for continuous molecular monitoring. Therefore, there is a critical need to develop refreshable biosensors that enable real-time, quantitative, continuous monitoring of protein biomarkers in complex body fluids. In this investigation, acute kidney injury (AKI) is employed as a disease model. Various protein biomarkers including tissue inhibitor of metalloproteinases-2, insulin-like growth factor-binding protein 7, and neutrophil gelatinase-associated lipocalin have been identified for early diagnosis/prognosis of AKI. Each biomarker has a specific time course in a specific setting of injury. Continuously monitoring a panel of AKI biomarkers relevant to physiologic and pathophysiologic processes in the injured tissue is very important, considering that AKI can evolve quickly. However, current protein quantification technologies cannot determine when an insult actually happens in the clinical setting, and fail to provide real-time evaluation of renal function for prompt clinical interventions. The long-term goal is to transform kidney disease treatment by developing a fully wireless, miniaturized, multifunctional biosensor that can be directly interfaced to a urinary catheter or the internal surface of patient bladder. The overall objective of this project is to design and develop a novel refreshable biosensor and to demonstrate continuous real-time monitoring of AKI protein biomarkers in patient urine. Specific aims include: 1. Design and realize biorecognition elements with high sensitivity, specificity and stability for refreshable biosensors. Based on feasibility studies, the biorecognition elements achieved by molecular imprinting of polymers exhibit high affinity to target protein biomarkers and are chemically, thermally, and mechanically stable under extreme conditions, which is critical for the proposed refreshable biosensor. 2. Design, develop and validate stimulators that locally and focally deliver surface acoustic energy to refresh biorecognition elements. The primary hypothesis is that locally and focally delivered high surface acoustic energy can desorb specifically bound target proteins and refresh the biorecognition sites of the biosensors. Such a new capability is highly valuable for monitoring the renal function of critically ill patients at risk of or with AKI, including patients with sepsis, shock, major surgery, and trauma.