Surface plasmon resonance spectroscopy for characterization of COVID-19 biosensors

动机在可靠、非常快速和稳健地检测包括COVID-19在内的冠状病毒方面存在重大技术差距。基于电化学的感测方法由于简单、快速响应、最小标签要求和低成本而对临床诊断有吸引力。靶cDNA和RNA与固定在电极上的DNA探针的结合引起氧化还原物质的传质变化并改变电流。影响生物传感器临床性能的因素还没有得到系统的研究。表面等离子体共振（SPR）光谱是生物传感器表征和用于确定目标分析物与表面固定的生物识别探针之间的结合亲和力的常用技术。SPR可以潜在地用于区分随机靶标与生物识别探针的非特异性结合和基质污染物在单层上的非特异性吸附。 目标。本项目的目的是开发和验证对SARS-CoV-2病毒RNA具有高亲和力和特异性的DNA探针。在该项目结束时，将开发有效的DNA探针和新的电化学传感方案，这些方案将适用于快速诊断COVID-19的电化学生物传感器。目标#1：DNA探针和靶分子的基于SPR的生物亲和性分析。目标#2：使用在目标#1中开发的优化的DNA探针开发电化学DNA生物传感器。 冲击使用合作伙伴的SPR技术开发和验证的DNA探针将被电化学生物传感器用于加拿大SARS-CoV-2的快速诊断。该项目开发的DNA探针和生物传感协议将有助于实施新的测试和生产策略，以提高下一代电化学生物传感器和诊断工具的质量控制。

Motivation. There is a critical technological gap in reliable, very rapid, and robust detection of coronaviruses, including COVID-19. Electrochemical-based sensing methods are appealing for clinical diagnostics, due to simplicity, fast response, minimum label requirement, and low cost. Binding of target cDNAs and RNAs to DNA probes immobilized on the electrode causes changes in the mass transfer of redox species and alters electrical currents. There are still factors affecting biosensors' clinical performance that have not been systematically investigated. Surface plasmon resonance (SPR) spectroscopy is a commonly used technique for biosensor characterization and for determining binding affinity between target analytes and surface-immobilized biorecognition probes. SPR can potentially be used to differentiate between non-specific binding of random targets to the biorecognition probe and non-specific adsorption of matrix contaminants on the monolayer. Aims. The objective of the proposed project is to develop and validate DNA probes with high affinity and specificity to viral RNA of SARS-CoV-2. At the end of this project, effective DNA probes and new electrochemical sensing protocols will be developed, which will be applicable in electrochemical biosensors for the rapid diagnosis of COVID-19. Aim #1: SPR-based bio-affinity analysis of DNA probes and target molecules. Aim #2: Electrochemical DNA biosensor development using optimized DNA probes developed in #Aim 1. Impact. The DNA probes developed and validated using the SPR technology of the partner will be used by electrochemical biosensors for the rapid diagnosis of SARS-CoV-2 in Canada. The DNA probes and biosensing protocols developed in this project will contribute to implementing new testing and production strategies to increase the quality control of the next generation of electrochemical biosensors and diagnostic tools.