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：
10611370
负责人：
Kirill A Afonin
金额：
$ 18万
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/10611370
关键词：
Address Affinity Antibodies Binding Binding Proteins Biological Biological Assay Biological Markers Cancer Diagnostics Characteristics Clinical Collaborations Complex Complex Mixtures DNA-Binding Proteins Detection Development Devices Dimensions Disease Early Diagnosis Elasticity Electronics 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 self assembly sensor technology simulation single molecule solid state synergism 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.

项目摘要这个跨学科的项目协同基本证明的三个研究小组的专业知识协同证明一种新的方法，可准确检测稀缺的蛋白质生物标志物。钥匙该方法的创新元素是使用线框样核酸纳米颗粒（NANP）结合直接或通过辅助抗体蛋白直接或高亲和力的蛋白靶标。蛋白质的结合通过首先将样品与Nanp的鸡尾酒一起孵育，然后使用A进行检查，然后使用A 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：离子电流和粒子停留时间在A的NANP时减少了一个特征量某些类型进入纳米孔。重要的是，通过将NANP的物理维度与物理匹配纳米孔的尺寸，我们期望大大增加核酸的居住类型纳米孔内的纳米颗粒，从而实现了超敏感的（亚曲面范围）检测蛋白质结合的生物标志物。通过设计我们的NANP探针以产生不同的离子特征。它们的蛋白质靶标，我们将使用相同的纳米孔进行多种蛋白质的多重检测以及通过将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三明治喜欢结构。该项目将由UNC Charlotte的Afonin Group进行，该项目将设计 Nanps，Nanps的东北大学Wanunu集团，将进行纳米孔检测实验，并 Aksimentiev组将使用建模技术的武器库来优化和改进检测战略。我们的超敏感，便携式，快速且潜在的低成本技术，用于定量蛋白质预计水平将在分析生物样品的分析中找到广泛的用途，最终提供敏感，可靠，以及对疾病指标生物标志物的最低侵入性鉴定，这可能是重要的创新癌症和其他疾病的早期诊断。