POINT OF CARE ACUTE KIDNEY INJURY DETECTION USING MOLECULARLY IMPRINTED GOLD NANOCAGES

项目描述(由申请人提供)：该项目的主要目标是设计和开发一种新的护理点检测方法，用于快速检测三种蛋白质生物标记物，即中性粒细胞明胶酶相关脂钙蛋白(NGAL)、肾脏损伤分子-1(Kim-1)和脂肪酸结合蛋白1(FABP1)，用于快速诊断和预测急性肾损伤(AKI)。在2000年至2009年间，美国需要透析的急性肾损伤的发病率每年上升约10%。急性肾损伤涉及肾脏排泄功能的迅速丧失。在同一时期，与需要透析的AKI相关的死亡人数增加了一倍多。在美国，每年约有120万患者被诊断为AKI，约30万人死于AKI。重症监护室的平均住院时间增加了3.5天，增加了大约90亿美元的成本。因此，血浆和/或尿液生物标记物水平的测定具有重要的临床意义。NGAL、白介素18(IL-18)、KIM-1、胱抑素C(CysC)和肝脏脂肪酸结合蛋白(FABP1)等蛋白被广泛研究为AKI的潜在生物标志物。例如，在AKI期间，尿NGAL水平增加了几个数量级，而血浆水平增加了5到10倍。已经认识到，仅血浆和尿液NGAL水平就可以早期发现有发生AKI风险的患者并预测其预后，并支持对已建立的AKI的鉴别诊断。目前的商业分析主要是基于免疫的，需要临床实验室来测量生理液体中的蛋白质生物标记物浓度，这不仅使分析成本高昂，而且会导致显著的延迟。我们建议开发一种新的基于血浆纳米换能器和人工抗体的护理点检测方法，用于检测生理和病理浓度的蛋白质生物标记物。具体地说，我们将研究金纳米笼作为等离子体纳米换能器和分子印迹人工抗体作为识别元件选择性捕获生物标志物，并用简单的手持式VIS-NIR光谱仪进行定量。除了显著降低成本外，设想的基于人造抗体的等离子生物传感器具有优异的化学和时间稳定性，无需特殊的存储条件(如冷藏、水合)，即使在资源有限的环境中也能够进行护理点生物标记物的定量。一旦这一早期的、创新的和探索性的研究完成，我们将为检测尿液和/或血浆中的蛋白质生物标志物奠定基础并部分开发高灵敏度的护理点式等离子生物传感器，从而实现AKI的快速筛查、早期检测、诊断和预后。将为下一步开发血浆纸基生物芯片奠定基础，该芯片用于监测尿液中的多个生物标志物，从而提高对各种病理状况的诊断和预后的准确性。

DESCRIPTION (provided by applicant): The primary objective of the project is to design and develop a novel point-of-care assay for rapid detection of three protein biomarkers, namely, neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1) and fatty acid binding protein 1 (FABP1), for rapid diagnosis and prognosis of acute kidney injury (AKI). The incidence of dialysis-requiring AKI, which involves rapid loss of kidney's excretory function, has risen about 10% per year in the United States between 2000 and 2009. During the same time period, the number of deaths associated with dialysis-requiring AKI has more than doubled. In the United States, about 1.2 million patients are diagnosed with AKI and about 0.3 million die of AKI each year. The average length of stay in an ICU is increased by 3.5 days adding about 9 billion dollars in cost. Thus, the measurement of plasma and/or urine biomarker levels is of huge clinical importance. Various proteins such as NGAL, interleukin-18 (IL-18), KIM-1, cystatin C (CysC), and liver fatty acid-binding protein (FABP1) have been extensively studied as potential biomarkers for AKI. For example, during AKI, urinary NGAL levels are increased by several log-orders of magnitude while plasma levels increase 5 to10-fold. It has been recognized that plasma and urine levels of NGAL alone provide early detection and prognosis of patients at risk of developing AKI and support the differential diagnosis of established AKI. Current commercial assays are primarily immune-based and require clinical lab settings to measure protein biomarker concentration in physiological fluids, which not only makes the assay expensive but also results in significant delay. We propose to develop a novel point-of-care assay based on plasmonic nanotransducers and artificial antibodies for the detection of protein biomarkers at physiological and pathological concentrations. Specifically, we will investigate gold nanocages as plasmonic nanotransducers and molecularly imprinted artificial antibodies as recognition elements for selective capture of biomarkers and their quantification by simple hand-held vis-NIR spectrometer. Apart from significantly lowering the cost, the envisioned plasmonic biosensor based on artificial antibodies offers excellent chemical and temporal stability, obviating the need for special storage conditions (e.g., refrigeration, hydration) of and enabling point-of-care biomarker quantification even in resource-limited settings. Once this early, innovative and exploratory research is completed, we will have laid the groundwork and partially developed highly sensitive point-of-care plasmonic biosensor for the detection of protein biomarkers in urine and/or plasma, which enables rapid screening, early detection, diagnosis and prognosis of AKI. The stage would be set for the next steps, of developing plasmonic paper-based biochips for monitoring multiple biomarkers from urine, which enhances the accuracy of diagnosis and prognosis of various pathological conditions.