权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Early diagnosis of acute kidney injury by aptasensors

适体传感器早期诊断急性肾损伤

基本信息

批准号：
8455374
负责人：
Muslum Ilgu
金额：
$ 15.05万
依托单位：
APTALOGIC, INC
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-15 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8455374
关键词：
Acute Renal Failure with Renal Papillary Necrosis Affect Affinity Antibodies Aptamer Technology Binding Bioinformatics Biological Assay Biosensor Chemicals Classification Creatinine Detection Development Devices Diabetes Mellitus Diagnosis Digestive System Disorders Early Diagnosis Early Intervention Early treatment Effectiveness Enzyme-Linked Immunosorbent Assay Evolution FDA approved Gelatinase A Goals Hour Immunoassay In Vitro Institutes Intensive Care Units Interferometry Kidney Diseases Label Lead Length Life Ligands Measurement Measures Medical Methods Modification Molecular Conformation Monitor National Institute of Diabetes and Digestive and Kidney Diseases Noise Nucleic Acids Output Patients Performance Phase Population Procedures Proteins RNA RNA Binding Renal function Reporting Research Sampling Serum Signal Transduction Silicon Speed Stress Surface System Temperature Thick Translating Triage Urea Urine Vision aptamer base cantilever chemical synthesis cost improved innovation instrument molecular marker mortality next generation sequencing nuclease outcome forecast patient population phase 1 study point of care prototype public health relevance receptor response tool

项目摘要

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. Our ultimate goal is 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 serum/urine neutrophil gelatinase-associated lipocalin (NGAL), an early marker of AKI [2-4 hours at the onset of AKI with cutoff concentration of ~ 150 ng/ml (6.8 nM)]. Aptamers are ssDNAs/RNAs that bind their targets avidly and specifically, and are selected in vitro by Systematic Evolution of 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 ¿m thick and 500-100 ¿m 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 this phase I study, we propose two specific aims: 1. Identify 2'F-modified aptamers that bind NGAL specifically with nanomolar affinity. 2. Demonstrate an aptamer-functionalized microcantilever device for NGAL detection. In phase I, we expect to develop a prototype of an aptamer-functionalized microcantilever that will be responsive to NGAL, which will be further optimized in phase II for reliable performance with serum and urine samples with robust signal/noise ratio, sensitivity, detection limit and dynamic range of measurement and that can be used at point of care to monitor for AKI.

描述（由申请人提供）：急性肾损伤（阿基）每年影响全球超过1000万人，不同患者人群的报告死亡率为15%至60%。然而，缺乏方法来支持阿基的早期诊断，这可能导致早期干预，改善治疗，改善预后和降低医疗成本。由于这些原因，开发阿基的早期诊断工具是NIDDK列出的征集研究主题之一。我们的最终目标是开发一种便携式设备，用于在护理点可靠地早期检测阿基。我们方案的创新在于将适体技术与血清/尿液中性粒细胞明胶酶相关脂质运载蛋白（NGAL）的微悬臂检测相结合，NGAL是阿基的早期标志物[阿基发作时2-4小时，截止浓度约为150 ng/ml（6.8 nM）]。适体是与其靶物强烈且特异性结合的ssDNA/RNA，并且通过指数富集配体系统进化（SELEX）在体外选择。对于生物传感器的发展，适体有许多优势，抗体，包括其较小的尺寸，化学合成和修饰的顺从性和适应性，包括微悬臂检测系统的广泛的测定格式。微悬臂梁是一种通常由硅制成的梁，其厚度为1-5 μ m，长度为500-100 μ m，仅在一端固定。当微悬臂梁的表面应力由于其表面上的配体-受体（例如适体-靶标）相互作用而发生构象变化而改变时，微悬臂梁弯曲。通过干涉测量法测量涂覆有不结合分析物的乱序核酸的参考悬臂和涂覆有结合分析物的适体的感测悬臂之间的表面应力差。在第一阶段的研究中，我们提出了两个具体目标：1。鉴定以纳摩尔亲和力特异性结合NGAL的2 'F修饰的适体。 2.展示用于NGAL检测的适体功能化微悬臂装置。在第一阶段，我们期望开发一种适体功能化的微悬臂梁的原型，该原型将对NGAL产生响应，其将在第二阶段进一步优化，以获得具有稳健的信噪比、灵敏度、检测限和动态测量范围的血清和尿液样品的可靠性能，并且可以在护理点用于监测阿基。