Nanobody-Based Electrochemical Biosensor for Real-Time Detection of Aerosolized SARS-CoV2

基于纳米抗体的电化学生物传感器，用于实时检测气溶胶 SARS-CoV2

• 批准号: 10320998
• 负责人: John R Cirrito
• 金额: $ 44.43万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-21 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320998
• 关键词: 2019-nCoV; Aerosols; Affinity; Air; Alzheimer' s Disease; American; Amino Acids; Amyloid beta-Protein; Antibodies; Area; Atmosphere; Binding; Biosensor; Blood; Breathalyzer Tests; Breathing; COVID-19; COVID-19 detection; COVID-19 test; Cell Nucleus; Cessation of life; Collection; Communities; Contact Tracing; Coronavirus; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Dimensions; Disease; Disinfection; Dust; Electrodes; Electrons; Environment; Environmental Risk Factor; Future; Goals; Hospitalization; Hour; Human; Humidity; Individual; Inhalation; Laboratories; Lead; Liquid substance; Llama; Longevity; Measurement; Measures; Membrane; Membrane Proteins; Methodology; Monitor; Mucous Membrane; Nebulizer; Nerve Degeneration; Nose; Observational Study; Particulate Matter; Performance; Persons; Pharyngeal structure; Physical condensation; Property; Protein Dynamics; Proteins; Rapid diagnostics; Recombinants; Reporting; Respiration; SARS coronavirus; SARS-CoV-2 antigen; SARS-CoV-2 transmission; Saliva; Sensitivity and Specificity; Signal Transduction; Soot; Specificity; Surface; Symptoms; Technology; Temperature; Testing; Tetrahydrocannabinol; Time; Touch sensation; Tube; Tyrosine; United States; Viral; Virus; aerosolized; air sampling; antigen test; atmospheric aerosols; atmospheric conditions; base; carbon fiber; community transmission; design; detection limit; detector; mortality; nanobodies; novel; oxidation; pandemic disease; particle; pathogen; pollutant; prototype; residence; respiratory; sensor; web site

ABSTRACT/PROJECT SUMMARY Coronavirus 2019 (COVID-19) has afflicted 6.2 million Americans and killed 190,000 as of early September 2020 (WHO website); a roughly 3% mortality. Between a shortage in testing and unidentified asymptomatic individuals, the actual number of those infected could be 6 to 24-fold higher than that reported. SARS-CoV-2 (CoV-2), the virus underlying the disease, results in a range of symptoms; in select cases a severe respiratory illness that impedes breathing that could lead to hospitalization and death. CoV-2 is transmitted person-to- person via inhalation of the virus through mucosal membranes of the nose and throat from transfer after touching a contaminated surface or by inhaling aerosolized virus. Unfortunately, COVID-19 is likely to be prevalent well into 2021 and beyond. We must increase our ability to test for CoV-2. First, testing is needed to diagnose individuals that are symptomatic or asymptomatic to reduce community spread. And second, monitoring gathering areas for airborne virus that could inform the decision to shutdown a space or implement disinfection and mitigation of an area. We propose to use an electrochemical biosensor in two detection devices, 1) a diagnostic breathalyzer for instant detection of CoV-2 and 2) an airborne detector for real-time, continuous surveillance of a large space. We have developed a novel ultra-sensitive, antibody-based electrochemical biosensor to detect CoV-2 repeat binding domain (RBD) spike protein. The technology is based on a micro-immunoelectrode (MIE) biosensor pioneered by the Cirrito laboratory to study protein dynamics in the setting of neurodegeneration (2,3). The biosensor uses voltammetry to measure the oxidation of tyrosine amino acids; oxidation is the release of electrons that the biosensor measures as a change in current. Antibodies are covalently attached to the electrode surface to provide selectivity. Our prototype CoV-2 biosensor is sensitive to 2 femtogram/ml, compared to several current CoV-2 antigen tests that are sensitive to the low picogram/ml range. The proposal will first (Aim 1) optimize our CoV-2 biosensor to detect CoV-2 viral particles, as well as test several parameters to increase sensitivity and longevity. Aim 2 will build a test breathalyzer that will utilize a nebulizer to generate virus laden air containing aerosol droplets similar to a breath that contain defined concentrations of CoV-2 viral particles. Aim 3 will test the airborne biosensor in a realistic environment. Co-I Chakrabarty’s laboratory has unique capabilities of mimicking real-world environmental conditions, especially in the context of atmospheric aerosols, necessary for testing and optimizing the biosensor’s performance for field deployment. Atmospheric conditions include relative humidity (RH) and temperature, as well as common airborne pollutants found indoors. Finding novel means to detect the CoV-2, as well as create a platform to detect other and future pathogens, would enable us to limit the viral spread throughout the community in the current and future pandemics.

摘要/项目摘要 2019年冠状病毒（Covid-19）遭受了620万美国人的折磨，截至9月初 2020（WHO网站）；大约3％的死亡率。在测试和未识别的无症状的短缺之间 个体，被感染者的实际数量可能比报告的人数高6至24倍。 SARS-CoV-2 （COV-2）是该疾病的基础病毒，导致一系列症状；在某些情况下，严重的呼吸道 阻碍呼吸的疾病可能导致住院和死亡。 COV-2是传播的人 - 通过吸入病毒通过鼻子和喉咙的粘膜通过转移而引起的人 触摸受污染的表面或吸入雾化病毒。不幸的是，Covid-19可能是 盛行到2021年及以后。 我们必须提高测试COV-2的能力。首先，需要进行测试以诊断 有症状或无症状以减少社区传播。其次，监视收集区域 空降病毒可以告知决定关闭空间或实施消毒的决定 一个区域。我们建议在两个检测设备中使用电化学生物传感器，1）诊断 呼吸分析仪，用于即时检测COV-2和2）实时，连续监视的空降探测器 一个大空间。 我们已经开发了一种新型的超敏感，基于抗体的电化学生物传感器来检测COV-2 重复结合结构域（RBD）尖峰蛋白。该技术基于微型免疫电极（MIE） Cirrito实验室率先研究神经变性的蛋白质动力学（2,3）。 生物传感器使用伏安法来测量酪氨酸氨基酸的氧化。氧化是释放 生物传感器测量电流变化的电子设备。抗体是共价附着在 电极表面以提供选择性。我们的原型COV-2生物传感器对2个fermtogram/ml敏感， 与当前对低皮克/mL范围敏感的当前COV-2抗原测试相比。 该提案将首先（AIM 1）优化我们的COV-2生物传感器，以检测COV-2病毒颗粒并进行测试 几个参数以提高灵敏度和寿命。 AIM 2将构建一个测试繁殖器，将使用 雾化器生成含有类似于呼吸的气溶胶液滴的病毒空气 COV-2病毒颗粒的浓度。 AIM 3将在现实的环境中测试机载生物传感器。 Co-i Chakrabarty的实验室具有模仿现实世界环境条件的独特功能，尤其是 在大气气溶胶的背景下，对于测试和优化生物传感器的性能所必需的 现场部署。大气条件包括相对湿度（RH）和温度以及常见 空气污染物在室内发现。 寻找新颖的方法来检测COV-2，并创建一个平台来检测其他和未来 病原体，将使我们能够在当前和未来限制整个社区的病毒传播 大流行。