A point-of-care ultrasensitive protein detection system

该项目的目标是开发一种用于超灵敏定量检测蛋白质的点护理（POC）传感系统。蛋白质是一大类生物分子，在许多临床和医疗保健应用中至关重要。目前蛋白质检测的定量方法都是基于实验室的。另一方面，现有的POC设备，虽然速度快，价格便宜，但不够灵敏，不能定量。因此，该项目的主要目标是：(a)开发用于检测非常小浓度蛋白质的传感器(b)开发可以提供蛋白质浓度定量测量的便携式系统。该项目的出发点是新型的基于适配体的蛋白质检测和定量技术，该技术最近由我们小组在电气和电子工程系生物启发技术中心开发。伦敦帝国理工学院。该项目的关键新颖之处在于开发了一种基于标准CMOS技术的新型传感器，该传感器将结合适体/DNA扩增，以实现低样本量的蛋白质检测。在这个项目中要回答的主要研究问题是：1。开发能够检测10-100pg/ml蛋白质水平的无标签CMOS传感器所需的标准化，优化和验证步骤是什么？在医疗点电子平台上实现传感器的技术和设计挑战是什么？拟议的项目将分三个主要步骤执行：首先，建立一个强大而有效的协议，以开发一种可访问、快速和可靠的无标签测试。关键任务将是实现一种基于微芯片的检测技术，该技术基于集成的化学传感器，如ISFET（离子敏感场效应晶体管）。2. 第二步是设计用于检测所需生物标志物的样品制备平台。这一步的挑战将是使用原始生物样本，如唾液，这将使开发非侵入性和用户友好的系统成为可能。3. 最后，第三步将是设计一个集成的电子护理点平台，能够量化所需的蛋白质。该平台将能够运行/控制DNA扩增和数据处理，目的是提供快速可靠的结果，因此可以在现实生活中轻松应用。该平台将允许基于定量蛋白质的诊断测试，用于即时检测，以改善许多应用中的医疗保健，并通过开发一种价格合理且超灵敏的新型检测方法来改变蛋白质组学领域。该平台的一个关键新颖之处是设计了一种电子芯片，能够在无标签和负担得起的系统中检测自由的互补DNA。这将有助于将基于实验室的概念验证技术转换为POC，小尺寸产品。另一个新颖之处在于开发适合POC应用的微流控平台的生化方案。进一步的创新将是设计一个具有新颖生物信号/数据处理能力的平台，这将导致基于适配体的创新具体到POC测试中。这项技术的应用非常广泛，从疾病检测到体重管理。该系统适用于任何需要快速、方便、可靠的诊断工具的应用。因此，该项目非常符合EPSRC临床技术雄心H3（医疗保健技术），旨在开发用于传感和诊断的新技术。此外，将这项技术发展成直接面向消费者的检测设备，通过直接向患者提供可获得的快速诊断系统，有可能改变社区卫生和护理（抱负H1）。

The aim of the project is to develop a Point-of-Care (POC) sensing system for ultrasensitive quantitative detection of proteins. Proteins are a large group of bio-molecules crucial in many clinical and healthcare applications. The current quantitative methods for protein detection are all lab-based. On the other side, the available POC devices, which although fast and inexpensive, are not sensitive enough and not quantitative. Therefore, the main objectives of this project are:(a) to develop sensors for detecting proteins in very small concentrations(b) to develop portable systems which can deliver quantitative measurements of protein concentrationsThe starting point of the project is the novel aptamer-based technique for protein detection and quantification which has recently been developed in our group at the Centre for Bio-Inspired Technology, Dept. of Electrical and Electronic Engineering, Imperial College London. The key novelty of the project is the development of a new sensor based on standard CMOS technology which will incorporate the aptamer/DNA amplification to enable low sample volume detection of proteins. The primary research questions to be answered in this project are:1. What are the standardisation, optimisation and validation steps necessary to develop a label-free CMOS sensor capable of detecting 10-100pg/ml protein levels?2. What are the technical and design challenges in implementing the sensor on a point-of-care electronic platform?The proposed project will be executed in three main steps:1. First, establishing a robust and efficient protocol for the development of an accessible, quick and reliable label-free test. The key task will be implementing a microchip-based detection technique, based on an integrated chemical sensor such as ISFET (Ion-Sensitive Field Effect Transistors). 2. The second step will be designing a platform for the sample preparation for the detection of the desired biomarker. Challenges in this step will be the use of raw biological samples such as saliva, which will enable the development of a non-invasive and user-friendly system. 3. Finally, the third step will be the design of an integrated electronic point-of-care platform able to quantify the desired proteins. This platform will be capable of running/controlling DNA amplification and data processing, with the aim of delivering quick and reliable results which can therefore find an easy placement in real-life applications. This platform will allow a quantitative protein-based diagnostic tests for Point-of-Care Testing to provide improved healthcare in many applications and to transform the area of proteomics with the development of an affordable and ultrasensitive novel assay.A key novelty of the platform is the design of an electronic chip capable of detecting free complementary DNA in a label-free and affordable system. This will facilitate translation of a lab-based proof-of-concept technique to a POC, small form factor product. Another novelty lies in developing a bio-chemical protocol with a microfluidic platform suitable for POC application. Further novelty will be the design of a platform with novel bio-signal/data processing capabilities, which will lead towards concretising the aptamer-based innovation into a POC test.The applications of the technology are numerous, from disease testing to weight management. This system can benefit any application which requires a quick, accessible and reliable diagnostic tool. Therefore, this project fits very well within the EPSRC Clinical Technologies Ambition H3 (Healthcare Technologies) aimed at the development of novel technologies for sensing and diagnosis. Moreover, the development of this technology into a direct-to-consumer testing device has the potential to transform community health and care (Ambition H1) by offering an accessible and fast diagnostic system directly to patients.