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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：
9144372
负责人：
Muslum Ilgu
金额：
$ 37.39万
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 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9144372
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 Diabetes Mellitus Diagnosis Digestive System Disorders Early Diagnosis Early Intervention Early treatment Effectiveness Evolution Funding Future Goals Health Hour In Vitro Institutes Intensive Care Units Interferometry Intervention Kidney Diseases 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 receptor research and development response sensor 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. 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.