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Multiplexed electronic counting of scarce protein targets using nucleic acid nanoparticles

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
批准号:
10611370
负责人:
Kirill A Afonin
金额:
$18.0万
依托单位国家:
美国
项目类别:
财政年份:
2022
资助国家:
美国
项目状态:
已结题
起止时间:
2022-05-01 至 2024-02-29

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中文摘要
翻译
项目摘要 这个跨学科的项目协同三个研究小组的专业知识,以证明原则 证明了一种新的方法,用于准确和灵敏地检测稀缺的蛋白质生物标志物。关键 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以高亲和力靶向蛋白。蛋白质结合 通过首先将样品与NANP的混合物一起孵育,然后使用 固态膜中的纳米孔。蛋白质检测依赖于测量流过的离子电流 纳米孔:当纳米孔的NANP为0.5 μ m时,离子电流和粒子停留时间降低特征量。 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,我们期望显著增加核酸的驻留类型 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。 蛋白质结合的生物标志物。通过设计我们的NANP探针,当结合到 他们的蛋白质目标,我们将实现多重检测的几种蛋白质物种使用相同的纳米孔, 以及通过将NANP探针和蛋白质生物标志物组装成生物标志物的组合检测。 三明治状结构。该项目将由位于夏洛特夏洛特的Afonin集团进行, NANP,东北大学的Wanunu小组将进行纳米孔检测实验, 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、快速和潜在低成本的蛋白质定量技术 水平预期将广泛用于生物样品的分析,最终提供灵敏,可靠, 和微创识别疾病指示生物标志物,这可能是重要的创新, 癌症和其他疾病的早期诊断。
英文摘要
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.
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