High-Throughput, Rapid, and Epitope-Specific Quantification of Neutralizing Antibodies Using Digital Nanoparticle Sensors

使用数字纳米颗粒传感器对中和抗体进行高通量、快速和表位特异性定量

• 批准号: 10432809
• 负责人: Chao Wang
• 金额: $ 22.9万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-20 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10432809
• 关键词: 2019-nCoV; ACE2; Address; Affinity; Antibodies; Antibody Therapy; Automation; Binding; Biological Assay; COVID-19; COVID-19 patient; Cells; Centrifugation; Cessation of life; Clinical; Color; Coronavirus; Country; Data; Data Analyses; Detection; Development; Disease; Ebola virus; Employment; Enzyme-Linked Immunosorbent Assay; Epitopes; Extinction (Psychology); Future; Gold; Herd Immunity; Human; Immune response; Immune system; Immunity; Immunization; Individual; Infection; Interdisciplinary Study; Laboratories; Light; Longitudinal Studies; Measurement; Measures; Metals; Monoclonal Antibodies; Mutation; Neutralizing antibody assay; Optics; Patients; Peptides; Persons; Plasma; Population; Precipitation; Process; Proteins; Public Health; RNA vaccine; Rapid diagnostics; Recombinants; Reporting; Research; Research Personnel; SARS coronavirus; SARS-CoV-2 B.1.617.2; SARS-CoV-2 antibody; SARS-CoV-2 antigen; SARS-CoV-2 immune response; SARS-CoV-2 infection; SARS-CoV-2 spike protein; SARS-CoV-2 variant; Sampling; Serum; Silver; Streptavidin; Suspensions; System; Technology; Testing; Time; Vaccination; Vaccines; Validation; Variant; Vendor; Viral; Virus; Virus Diseases; antibody detection; antibody test; base; convalescent plasma; coronavirus disease; cost; cross reactivity; design; detection limit; diagnostic platform; digital; effectiveness evaluation; efficacy study; first responder; health economics; high reward; high throughput screening; human coronavirus; nanoparticle; nanorod; nanosensors; neutralizing antibody; novel; novel coronavirus; novel virus; operation; particle; pathogen; pathogenic virus; population based; portability; rapid detection; receptor binding; scale up; sensor; serosurveillance; skills; transmission process; vaccine candidate; vaccine efficacy; variants of concern

Project Summary/Abstract As of mid June 2021, the new coronavirus disease (COVID-19), caused by SARS-CoV-2 virus, has infected ~180 million people and causing ~3.8 million deaths globally. There are more than 34 million confirmed cases and ~620,000 deaths in the U.S alone. The fast transmission, asymptotic infection in some individuals, currently still limited supplies of vaccines in many countries, and constant virus mutation have made COVID-19 an unprecedented global threat to human health and economics. Neutralizing antibodies (nAbs) recognize SARS- CoV-2’s spike protein and block cellular entry, acting as the first responders in the immune system towards pathogen clearance. Evaluating the effectiveness of nAbs against viral pathogens is important in understanding the level and duration of sterilizing immunity after natural infection or following vaccination, particularly given the rise of novel variants with vaccine escape potential. However, many of the available neutralizing assays used to assess nAb function involve propagation of viruses and thus require such assays be conducted in a biosafety level 3 (BSL3) lab settings, which, unfortunately, is unavailable to many researchers or clinicians. In addition, clinical laboratory-based antibody tests measure the total Ab level responding to SARS-CoV-2 antigens, without functionally evaluating pathogen-bound Abs and therefore cannot predict neutralizing activity. To bridge these technological gaps, we propose a multidisciplinary research plan to address the fundamental challenges in low- cost, high-throughput, fast, simple, and quantitative assay format in studying COVID-19 immune response. The investigators at ASU with complementary expertise in nanosensor design, antibody characterization, and coronaviruses will collaboratively develop a new and high-reward research strategy to establish a metal nanoparticle (MNP)-based nAb assay platform. This platform presents a few key features distinguishing it from previous technologies. First, the MNP assays are quantitative and accurate, with an expected dynamic range of 3 to 4 logs and a detection limit in the picomolar range. Further, this assay can be implemented in a rapid detection format without any washing steps, thus significantly simplifying its operation, reducing assay time to a few minutes, and making it feasible for mass-testing. Importantly, the assay is capable of detecting virus variants by targeted binding to the epitopes on the spike protein that are sensitive to mutations. Additionally, the readout can be performed in a well plate compatible with high-throughput screening with added portable electronic components, making the system automated in both detection and data analysis. We envision the employment of this rapid and quantitative nAb assay will also help timely determine the potential best uses of convalescent plasma and antibody treatment with future emerging novel viruses. Its low cost, simple operation, and automation capability are also very useful in longitudinal studies of the immune response related to COVID-19 infection, vaccination, and potential viral escape due to mutations. The nAb sensing can also be used for large-population sero-surveillance in determining the level of population-based immunity (herd immunity) against any virus strains.

项目概要/摘要 截至 2021 年 6 月中旬，由 SARS-CoV-2 病毒引起的新型冠状病毒病（COVID-19）已感染 全球约 1.8 亿人死亡，约 380 万人死亡。确诊病例超过 3400 万例。 目前，仅在美国就有约 62 万人死亡，某些人的传播速度很快，呈无症状感染。 许多国家的疫苗供应仍然有限，并且不断的病毒变异使COVID-19成为 中和抗体（nAb）对人类健康和经济构成前所未有的全球威胁。 CoV-2 的刺突蛋白可阻断细胞进入，充当免疫系统中的第一反应者 评估 nAb 对病毒病原体的有效性对于理解病原体很重要。 自然感染或疫苗接种后的灭菌免疫力的水平和持续时间，特别是考虑到 具有疫苗逃逸潜力的新变体的兴起然而，许多可用的中和测定用于 评估 nAb 功能涉及病毒的繁殖，因此需要在生物安全环境下进行检测 不幸的是，许多研究人员或精英无法获得 3 级（BSL3）实验室设置。 基于临床实验室的抗体测试可测量对 SARS-CoV-2 抗原做出反应的总抗体水平，无需 功能评估病原体结合抗体，因此无法预测中和活性。 技术差距，我们提出了一项多学科研究计划，以解决低技术领域的根本挑战 研究 COVID-19 免疫反应的低成本、高通量、快速、简单和定量的检测形式。 亚利桑那州立大学的研究人员在纳米传感器设计、抗体表征和 冠状病毒将合作开发一项新的高回报研究策略，以建立金属 该平台具有一些区别于纳米颗粒 (MNP) 的 nAb 检测平台的关键特征。 首先，MNP 检测是定量且准确的，预期动态范围为 3 至 4 个对数和皮摩尔范围内的检测限此外，该测定可以快速实施。 检测形式无需任何清洗步骤，从而显着简化其操作，将检测时间缩短至 几分钟，使其可以进行大规模测试，重要的是，该检测能够检测病毒变体。 通过靶向结合对突变敏感的刺突蛋白表位。 可以在与高通量筛选兼容的孔板中进行，并添加便携式电子设备 组件，使系统在检测和数据分析方面实现自动化。 这种快速、定量的 nAb 测定也将有助于及时确定恢复期的潜在最佳用途 血浆和抗体治疗未来新兴的新型病毒，其成本低、操作简单、自动化。 能力对于与 COVID-19 感染相关的免疫反应的纵向研究也非常有用， 疫苗接种以及由于突变而导致的潜在病毒逃逸 nAb 传感也可用于大量人群。 血清监测以确定针对任何病毒株的人群免疫（群体免疫）水平。