Multiplexed electronic counting of scarce protein targets using nucleic acid nanoparticles

使用核酸纳米粒子对稀有蛋白质靶标进行多重电子计数

基本信息

批准号：
10353490
负责人：
Kirill A Afonin
金额：
$ 24.33万
依托单位：
UNIVERSITY OF NORTH CAROLINA CHARLOTTE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10353490
关键词：
Address Affinity Antibodies Binding Binding Proteins Biological Biological Assay Biological Markers Cancer Diagnostics Characteristics Clinical Complex Complex Mixtures DNA-Binding Proteins Detection Development Devices Dimensions Disease Early Diagnosis Elements Emerging Communicable Diseases Enzyme-Linked Immunosorbent Assay Event Foundations Incubated Individual Laboratories Libraries Life Liquid substance Literature Measurement Measures Membrane Microscopic Modeling Nanostructures Nanotechnology Nucleic Acids Pathogenicity Patient Care Proteins RNA-Binding Proteins Research Research Personnel Research Proposals Sampling Shapes Signal Transduction Specificity Structure Techniques Technology Time Tissues Universities antibody conjugate aptamer clinical application combinatorial cost cost effective design dimer electric field experimental study high reward high risk improved individual patient innovation instrument minimally invasive multiplex detection nano nanoparticle nanopore novel strategies particle personalized medicine portability prenatal protein biomarkers rapid diagnosis residence response sensor technology simulation single molecule solid state technology development virtual

项目摘要

PROJECT SUMMARY This interdisciplinary project synergizes the expertise of three research groups for a proof-of-principle demonstration of a novel approach for accurate and sensitive detection of scarce protein biomarkers. The key innovative element of the approach is the use of wireframe-like nucleic acid nanoparticles (NANPs) to bind protein targets with high affinity either directly or by means of auxiliary antibody proteins. Binding of the protein targets is detected by first incubating the sample with a cocktail of NANPs and then examining them using a nanopore in a solid-state membrane. Protein detection relies on measurement of the ionic current flowing through the nanopore: the ionic current and particle dwell time decreases by a characteristic amount when a NANP of a certain type enters the nanopore. Importantly, by matching the physical dimensions of the NANP to the physical dimension of the nanopore, we expect to dramatically increase the residence type of the nucleic acid nanoparticles within the nanopore and thereby achieve ultra-sensitive (sub-picomolar range) detection of the protein-bound biomarkers. By designing our NANP probes to produce distinct ionic signatures when bound to their protein targets, we will achieve multiplex detection of several protein species using the same nanopore as well as combinatorial detection of biomarkers by assembling the NANP probes and protein biomarkers into a sandwich like structures. The project will be carried out by the Afonin group at UNC Charlotte that will design NANPs, the Wanunu group at Northeastern University that will perform the nanopore detection experiments, and the Aksimentiev group that will use an arsenal of modeling techniques to optimize and improve the detection strategy. Our ultra-sensitive, portable, rapid, and potentially low-cost technology for quantification of protein levels is expected to find broad use for the analysis of biological samples, eventually offering sensitive, reliable, and minimally invasive identification of disease-indicative biomarkers that could be important innovations for early-stage diagnostics of cancer and other diseases.

项目概要这个跨学科项目整合了三个研究小组的专业知识，以进行原理验证展示了一种准确、灵敏地检测稀有蛋白质生物标志物的新方法。关键该方法的创新要素是使用线框状核酸纳米颗粒（NANP）来结合直接或通过辅助抗体蛋白以高亲和力靶向蛋白质。蛋白质的结合首先将样品与 NANP 混合物一起孵育，然后使用固态膜中的纳米孔。蛋白质检测依赖于流过的离子电流的测量纳米孔：当 NANP 为某些类型进入纳米孔。重要的是，通过将 NANP 的物理尺寸与物理尺寸相匹配纳米孔的尺寸，我们期望显着增加核酸的停留类型纳米孔内的纳米颗粒，从而实现超灵敏（亚皮摩尔范围）的检测蛋白质结合生物标志物。通过设计我们的 NANP 探针，在结合时产生不同的离子特征他们的蛋白质目标，我们将使用与以及通过将 NANP 探针和蛋白质生物标志物组装成一个生物标志物的组合检测类似三明治的结构。该项目将由北卡罗来纳大学夏洛特分校的 Afonin 小组实施，该小组将设计 NANPs，东北大学的 Wanunu 小组将进行纳米孔检测实验，以及 Aksimentiev 小组将使用一系列建模技术来优化和改进检测战略。我们的超灵敏、便携式、快速且潜在低成本的蛋白质定量技术水平预计将广泛用于生物样品分析，最终提供灵敏、可靠、以及疾病指示生物标志物的微创鉴定，这可能是重要的创新癌症和其他疾病的早期诊断。