Early diagnosis of acute kidney injury by aptasensors

适体传感器早期诊断急性肾损伤

• 批准号: 9338228
• 负责人: Muslum Ilgu
• 金额: $ 30.64万
• 依托单位: APTALOGIC, INC
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9338228
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Affect; Affinity; Antibodies; Aptamer Technology; Binding; Biological Assay; Biological Markers; Biosensor; Blood; Blood Proteins; Characteristics; Chemicals; Classification; Clinics and Hospitals; Cost Analysis; Creatinine; Detection; Development; Device or Instrument Development; Devices; Diagnosis; Early Diagnosis; Early Intervention; Early treatment; Effectiveness; Evolution; Funding; Future; Goals; Hour; In Vitro; Intensive Care Units; Interferometry; Intervention; LCN2 gene; Label; Laboratories; Lead; Length; Life; Ligands; Measurement; Measures; Medical; Methods; Modification; Molecular Conformation; Monitor; National Institute of Diabetes and Digestive and Kidney Diseases; Noise; Nucleic Acids; Onset of illness; Output; Patient Monitoring; Patients; Performance; Phase; Plasma; Population; Procedures; Process; Proteins; Quantitative Evaluations; RNA; RNA Binding; Renal function; Reporting; Research; Risk; Role; Sales; Sampling; Serum; Serum Proteins; Signal Transduction; Silicon; Single-Stranded DNA; Sodium Chloride; Specificity; Stress; Surface; System; Technology; Temperature; Thick; Time; Urea; Urine; Vision; aptamer; base; cantilever; chemical synthesis; cost; improved; innovation; instrument; miniaturize; monitoring device; mortality; nanomolar; outcome forecast; patient population; phase 1 study; phase 2 study; phase 3 study; point of care; portability; public health relevance; receptor; research and development; response; sensor; tool

 DESCRIPTION (provided by applicant): Acute kidney injury (AKI) affects more than 10 million people worldwide each year with reported mortalities from 15 to 60% in different patient populations. However, methods are lacking to support the early diagnosis of AKI that could lead to early intervention, improved therapy, better prognosis, and lower medical costs. For these reasons, the development of early diagnosis tools for AKI is one of the solicited research topics listed by the NIDDK. Neutrophil gelatinase-associated lipocalin (NGAL) is a reliable biomarker for AKI and may also have some predictive uses. We propose to develop a portable device for reliable early detection of AKI at point-of-care. The innovation of our proposal lies in the combination of aptamer technology with microcantilever detection of NGAL, an early marker of AKI [2-4h at the onset of AKI; cutoff ~150 ng/ml]. Aptamers are single stranded DNAs or RNAs that bind their targets avidly and specifically, and are selected in vitro by Systematic Evolution f Ligands by Exponential enrichment (SELEX). For biosensor development, aptamers have many advantages over antibodies, including their smaller size, amenability to chemical synthesis and modifications and adaptability to a broad range of assay formats including a microcantilever detection system. A microcantilever is a beam, often of silicon, that is 1-5 µm thick and 500-100 µm long and anchored only at one end. The microcantilever bends when its surface stress changes due to the conformation change that occurs upon ligand-receptor (e.g. aptamer-target) interaction on its surface. The differential surface stress between a reference cantilever coated with a scrambled nucleic acid that does not bind the analyte and a sensing cantilever coated with an aptamer that binds the analyte is measured by interferometry. In a phase I study, we isolated several aptamers that bind NGAL and demonstrated the abilities of one of these to sense NGAL when the aptamer is attached to a microcantilever. In this phase II study, we propose the following specific aims: 1) Compare several aptamers for specificity, functionality in the presence of serum proteins, affinity for NGAL, effectiveness in the presence of salt mixtures characteristic of plasma and urine, and compatibility with the microcantilever device, 2) Optimize the best performing aptamers for length, sequence, stability and optimal assay performance in blood and urine, and 3) Optimize the microcantilever device for aptamer sensing (dynamic range of 1-100 nM for detection, signal/noise ratio > 10, covariance < 10%). Miniaturize the microcantilever device to a size equal to or smaller than a shoebox. From these studies, we expect to develop an optimized and reliable aptamer-functionalized microcantilever by which NGAL can be quantified in plasma and urine samples with robust signal/noise ratio, sensitivity, detection limit and dynamic range of measurement. Phase III studies will involve manufacturing and validating the device for monitoring NGAL to detect AKI at point-of-care.

 描述（由适用提供）：急性肾脏损伤（AKI）每年影响全球超过1000万人，报告的死亡人数为15％至60％。但是，缺乏支持AKI早期诊断的方法，这可能会导致早期干预，改善治疗，更好的预后和降低医疗费用。由于这些原因，为AKI的早期诊断工具的开发是NIDDK列出的巩固研究主题之一。中性粒细胞明胶酶相关的Lipocalin（NGAL）是AKI的可靠生物标志物，也可能具有一些预测用途。我们建议开发一种便携式设备，以在护理点上可靠地对AKI的可靠检测。我们提案的创新在于Atamer技术与MicroCantilever检测Ngal的结合，NGAL是AKI的早期标志[AKI发作时的2-4H；截止〜150 ng/ml]。 ATAMER是单个绞合的DNA或RNA，其靶标和特定地结合其靶标，并且通过指数富集（SELEX）在体外通过系统的进化F配体进行体外选择。对于生物传感器的开发，适体比抗体具有许多优势，包括其较小的尺寸，化学合成的舒适性以及对包括微型管理器检测系统在内的广泛测定格式的适应性。微型磁管是硅的光束，通常是1-5 µm厚，长500-100 µm，仅在一端固定。当微型管理器的表面应力因其表面上的配体增强器（例如apatmer-target）相互作用而发生的会议变化而发生变化时，其表面应力发生变化。通过干涉测量法测量了与分析物覆盖的参考悬臂之间的差分表面应力，该参考悬臂覆盖了不结合分析物的炒核酸和涂有结合分析物的APATMER的感应悬臂。在I阶段研究中，我们分离了几个结合NGAL的适体，并证明了其中一种适合剂时，当适体连接到微型管理员时。 In this phase II study, we propose the following specific aims: 1) Compare several aptamers for specificity, functionality in the presence of serum proteins, affinity for NGAL, effectiveness in the presence of salt mixes characteristic of plasma and urine, and compatibility with the microcantilever device, 2) Optimize the best performing aptamers for length, sequence, stability and optimal assay performance in blood and urine, and 3) Optimize the用于ATAMER感应的微型磁管设备（用于检测的1-100 nm的动态范围，信号/噪声比> 10，协方差<10％）。将微型管理器设备的小型化度达到等于或小于鞋盒的尺寸。从这些研究中，我们期望开发出优化且可靠的适体官能化的微型抗体，通过该适用性官能化的微型抗体，可以在血浆和尿液样品中量化具有强大信号/噪声比，灵敏度，检测极限和测量动态范围的尿液样品。第三阶段的研究将涉及制造和验证该设备，以监视NGAL以在护理点检测AKI。