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.

项目摘要/摘要 截至2021年6月中旬，由SARS-COV-2病毒引起的新冠状病毒病（COVID-19）已感染 约1.8亿人，全球造成约380万人死亡。有超过3400万个已确认的案件 仅在美国，约有62万人死亡。快速传播，某些个体中的渐近感染，目前 在许多国家，仍然有限的疫苗供应，恒定的病毒突变使Covid-19成为 空前对人类健康和经济学的全球威胁。中和抗体（NABS）识别的SARS- COV-2的尖峰蛋白和阻止细胞进入，充当免疫系统中的第一反应者 病原体清除。评估NABS针对病毒病原体的有效性对于理解很重要 自然感染或疫苗后，尤其是考虑到疫苗后的消毒免疫的水平和持续时间 具有疫苗逃生潜力的新型变体的兴起。但是，许多可用的中和测定法 评估NAB功能涉及病毒的传播，因此需要在生物安全中进行此类评估 第3级（BSL3）实验室设置，不幸的是，许多研究人员或临床医生无法获得。此外， 基于临床实验室的抗体测试测量对SARS-COV-2抗原反应的总AB水平，没有 在功能上评估结合病原体的ABS，因此无法预测中和活性。桥接这些 技术差距，我们提出了一项多学科研究计划，以应对低 - 在研究COVID-19的免疫反应中，成本，高通量，快速，简单和定量评估格式。这 ASU的调查人员具有纳米传感器设计，抗体表征和 冠状病毒将共同制定一种新的高级研究策略来建立金属 纳米颗粒（MNP）基于NAB测定平台。该平台提供了一些与众不同的关键功能 以前的技术。首先，MNP分析是定量和准确的，预期的动态范围为 在皮莫尔范围内的3至4个原木和检测极限。此外，该测定可以快速实施 检测格式没有任何洗涤步骤，因此大大简化了其操作，将测定时间缩短为 几分钟，使其可用于大规模测试。重要的是，该测定能够检测病毒变体 通过靶向与突变敏感的尖峰蛋白上的表位结合。另外，读数 可以在兼容的井板中与高通量筛选兼容，并添加便携式电子 组件，使系统在检测和数据分析中都自动化。我们设想就业 在这种快速而定量的NAB测定中，还将有助于及时确定康复的最佳用途 血浆和抗体治疗未来新兴病毒。其低成本，简单的操作和自动化 能力在与Covid-19感染有关的免疫反应的纵向研究中也非常有用。 疫苗接种和由于突变引起的潜在病毒逃生。 NAB感应也可以用于大型人口 在确定对任何病毒菌株的基于人群的免疫组织化学（HERD免疫组织化学）的水平时，血清均应监测。