NISED - A Nanoparticle Immunoassay based on Single-entity Electrochemical Detection

NISED - 基于单实体电化学检测的纳米颗粒免疫分析

• 批准号: 446370753
• 负责人: Professor Dr. Bernhard Wolfrum
• 金额: --
• 依托单位: Professur für Neuroelektronik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/446370753?language=en
• 关键词: NISED; Nanoparticle; Immunoassay; based; Single

The aim of this proposal is to investigate a new analytical technique for rapid and selective quantification of biomolecules using single-entity electrochemical detection on a chip. We will make use of thiol-linked silver nanoparticles with recognition molecules including antibodies and aptamers for specifically binding target molecules in solution. The sensor system will comprise an array of closely spaced and individually addressable electrodes for capturing and detecting nanoparticles. Some of the electrodes will be functionalized with recognition molecules via thiol-gold interactions to serve as capture probes for the target molecules during a preliminary incubation phase with the analyte. After incubation and exposure to functionalized nanoparticles, a reductive pulse will be applied to the capture electrodes to release bound nanoparticles by breaking the metal-sulfur bonds. The released particles will subsequently be detected at adjacent detection electrodes via impact electrochemistry. During this process, the silver nanoparticles will be oxidized generating short and defined current spikes. As the frequency of nanoparticle impacts depends on the mass transport of particles to the detection electrode, the rate of recorded current spikes will reflect the number of captured nanoparticles and the concentration of the target molecules in the analyte. A microfluidic confinement will ensure a detection efficiency of the nanoparticles close to 100%. Overall, the proposed concept will provide a highly sensitive immunoassay with electronic readout based on single-entity nanoparticle detection. As a proof of concept we will investigate the performance of this assay for the selective detection of insulin in serum as well as whole blood.

这项建议的目的是研究一种新的分析技术，利用芯片上的单一实体电化学检测来快速和选择性地定量生物分子。我们将利用硫醇连接的银纳米颗粒与识别分子(包括抗体和适配子)特异性地结合溶液中的目标分子。传感器系统将包括一组间隔紧密且可单独寻址的电极，用于捕获和检测纳米颗粒。一些电极将通过硫醇-金相互作用与识别分子进行功能化，以在与分析物的初步孵育阶段充当目标分子的捕获探针。在孵化和暴露于功能化纳米颗粒后，将在捕获电极上施加还原脉冲，通过打破金属-硫键来释放结合的纳米颗粒。释放的颗粒随后将通过撞击电化学在相邻的检测电极上被检测到。在这个过程中，银纳米粒子将被氧化，产生短而明确的电流尖峰。由于纳米颗粒撞击的频率取决于颗粒到检测电极的质量传输，记录的电流尖峰的速率将反映捕获的纳米颗粒的数量和目标分子在分析物中的浓度。微流控约束将确保纳米颗粒的检测效率接近100%。总体而言，拟议的概念将提供一种基于单一实体纳米颗粒检测的具有电子读数的高灵敏免疫分析。作为概念验证，我们将研究该方法在选择性检测血清和全血中胰岛素的性能。