Nanopore Sensors for Multiplexed Analysis of DNA in Clinically Relevant Samples

纳米孔已成为DNA、RNA和蛋白质单分子分析的通用传感技术。单个分子是通过测量离子电流的变化来检测的，因为它们通过一个纳米级的孔，分隔两个含有盐溶液的房间。然而，为了通过纳米孔传感主要阶段的初步证明，有几个关键领域需要改进。该项目将研究开发一种强大的纳米孔检测方法，用于对复杂的临床相关样本中的不同生物分子进行多重检测。该项目将需要在几个方面进行创新，包括重新设计和优化在复杂溶液中进行传感的传感系统，调查纳米孔功能化对传感器性能的好处，以及概念验证测试同时检测和量化临床样本中的多个生物标志物。该项目将研究和开发用于计算生物标志物和基因的传感协议。由于纳米孔传感技术允许对与DNA结合的分子进行一般计数和大小调整，该项目还将致力于将传感器用于分析结合蛋白质质量的限制提高到亚kDa。后者与识别蛋白质修饰和小配体结合事件有关，并应允许对活体的工作方式进行独特的洞察。

Nanopores have become a versatile sensing technique for the single molecule analysis of DNA, RNA and proteins. Single molecules are detected by measuring changes in ionic current flow as they pass through a nanometre scale pore separating two chambers containing salt solution. However, to advance past an initial proof of principal stage for nanopore sensing, several key areas need to be improved. This project will investigate the development of a robust nanopore sensing assay for multiplexed detection of diverse biomolecules in complex clinically relevant samples. The project will require innovation on several fronts, including redesigning and optimizing the sensing system for sensing in complex solutions, investigating benefits of functionalizing the nanopores for sensor performance, and proof of concept tests detecting and quantifying multiple biomarkers concurrently in clinical samples. The project will investigate and develop sensing protocols for counting biomarkers and genes. As the nanopore sensing technique allows for the general counting and sizing of molecules binding to DNA, the project will also aim to push the limits to use the sensors for analysing the mass of bound proteins down to sub kDa. The latter is relevant to identify protein modification and small ligand binding events and should allow for unique insights into the workings of living organisms.