Refreshable Biosensors for Continuous Renal Function Monitoring

项目摘要 拟议的探索性研究的目标是设计和开发新型的可刷新生物传感器， 克服与现有生物传感器相关的关键根本挑战，实现实时、定量、 持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 还有汗水。常规技术，如酶联免疫吸附试验(ELISA)、Western Blot和 质谱学使生物样品中蛋白质的发现和研究成为可能。这些传统的 由于生物识别的稳定性差，生物检测通常仅限于一次性或短时间使用 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重复使用。 这些传统的工艺与可穿戴和可植入的连续分子设备不兼容 监控。因此，迫切需要开发能够实时、 定量、连续监测复杂体液中的蛋白质生物标记物。在这项调查中，急性 肾损伤是一种疾病模型。包括组织抑制因子在内的各种蛋白质生物标志物 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结合蛋白 已被确定为AKI的早期诊断/预后。每个生物标记物都有特定的时间进程 伤情的设定。持续监测一组与生理和病理生理相关的AKI生物标志物 考虑到AKI可以快速演变，损伤组织中的过程非常重要。但是，当前 蛋白质定量技术无法确定侮辱在临床环境中实际发生的时间，以及 未能提供实时的肾功能评估，以便及时进行临床干预。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来改变肾脏疾病的治疗方法 可直接连接到导尿管或患者膀胱内表面。总的目标是 本项目旨在设计和开发一种新型的可刷新生物传感器，并展示连续的实时 患者尿液中AKI蛋白生物标志物的监测。具体目标包括：1.设计并实现生物识别 用于可刷新生物传感器的具有高灵敏度、特异度和稳定性的元件。根据可行性研究， 分子印迹聚合物获得的生物识别元件对靶蛋白具有很高的亲和力 生物标志物，在极端条件下在化学、热和机械上是稳定的，这对 建议的可刷新生物传感器。2.设计、开发和验证本地和集中交付的刺激器 用于刷新生物识别元件的表面声能。主要的假设是局部和局部的 提供的高表面声波能量可以解吸特定结合的目标蛋白并刷新 生物传感器的生物识别位置。这种新的能力对监测肾功能具有很高的价值。 有AKI风险的危重患者，包括败血症、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.