Rapid SARS-CoV-2 Spike Protein Neutralizing Antibody Tests for Evaluating COVID Vaccine-Generated Immunity

摘要/摘要： 新冠肺炎疫苗的提供是结束这一大流行并使生活恢复正常的重要一步。 成功的疫苗接种应诱导足够滴度的特异性中和抗体(NAB)，以对抗严重的 预防新冠肺炎感染SARS冠状病毒2型。SARS-CoV-2病毒尖峰 蛋白质nabs测试将是验证接受者是否有足够的保护和确定 豁免期。常规SARS-CoV-2刺突蛋白NABS试验，如空斑减少中和 检测和酶联免疫吸附试验都是以实验室为基础的，不适合大规模测量。而侧向流动 检测可以提供廉价和快速的检测，现有的侧向流动测试条不能提供准确的定量 测量。在这个SBIR项目中，DL ADV-Tech计划开发一种高度敏感、可靠和 S-非典冠状病毒蛋白NABS快速检测仪测定NABS浓度。少校 该方法的意义在于能够准确、快速和大规模地量化SARS-CoV-2 S-利用我们先进的介孔纳米花(MNFS)信号放大技术进行蛋白质NABS (正在申请专利)和基于智能手机的读出技术。最近我们发现了一类新的催化剂 纳米材料，双金属铂-钯MNFS，具有令人印象深刻的类似过氧化物酶的催化活性和良好的稳定性 在恶劣的条件下。我们正在申请专利的“基于MNFS的双金属横向流动免疫分析”展示了 与商业胶体金标记物相比，PT-Pd MNFS可以将灵敏度提高1000倍。巨大的 多国框架的活动对于使拟议的测试平台更具说服力具有特别重要的意义 以提高快速分析的准确性。同时，侧向流动带的色度结果将 使用智能手机读数获取和分析，这允许快速和准确地量化 SARS-CoV-2 S蛋白NAB浓度。与智能手机集成，该测试设备将提供 定量测量，易于操作，并允许实时数据收集和共享。建议数 检测装置可实现对SARS-CoV-2 S蛋白的高度可靠、简单、可靠和定量的检测 血中NABS，为准确密切监测SARS-CoV-2 S-NABS蛋白水平提供了强有力的工具 大规模的人口规模，并为个人和临床医生提供了更多的信心来抗击这一流行病。 在第一阶段，重点是验证SARS-CoV-2 S-蛋白NABS快速定量的概念 提出了检测装置，并从以下方面论证了该检测装置在血液样本中的可行性 灵敏度、准确度和检测范围。铂-钯MNFS增强型智能手机平台预计将拥有 高准确度(5%RSD)的SARS-CoV-2 S蛋白NABS定量(1 ng/ml-100ug/ml)。在第二阶段， 测试将使用临床样本进行评估，并针对实际应用进行进一步优化。建议数 检测设备一旦得到验证，将极大地造福于美国和世界各地的公共健康。有一个 如此有效的测试设备具有巨大的市场和巨大的商业机会。 1

Summary/Abstract: The availability of COVID-19 vaccines is a significant step in ending this pandemic and returning life to normal. Successfully vaccination should induce sufficient titers of specific neutralizing antibodies (nAbs) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to prevent infection of COVID-19. SARS-CoV-2 spike protein nAbs test will be essential to verify whether the recipients have sufficient protection and determine the duration of immunity. Conventional SARS-CoV-2 spike protein nAbs test, such as plaque reduction neutralization test and ELISA, are laboratory based, which are not suitable for large scale measurement. While lateral flow assays can provide cheap and quick tests, current lateral flow test strips can’t provide accurate quantitative measurement. In this SBIR project, DL ADV-Tech proposes to develop a highly sensitive, reliable and rapid SARS-CoV-2 S-protein nAbs testing device for quantification of nAbs concentrations. The major significance of this proposed approach is the ability to accurately, quickly and massively quantify SARS-CoV-2 S-protein nAbs by leveraging our advanced mesoporous nanoflowers (MNFs) signal amplification technique (patent pending) and smartphone-based readout technique. Recently we discovered a new class of catalytic nanomaterials, bimetal Pt-Pd MNFs, with impressive peroxidase-like catalytic activity and excellent stability under harsh conditions. Our patent pending “bimetal MNFs-based lateral flow immunoassays” demonstrated Pt-Pd MNFs can improve the sensitivity by 1000 times over commercial colloidal gold labels. The tremendous activity of MNFs is of particular importance to make this proposed test platform substantially more compelling for improvement of accuracy in rapid analysis. Meanwhile, the chromatic results from the lateral flow strips will be acquired and analyzed using a smartphone readout, which allows rapid and accurate quantification of SARS-CoV-2 S-protein nAb concentrations. Integrated with smartphone, this testing device will provide quantitative measurements and easy operation, and allow real-time data collection and sharing. The proposed testing device allows highly reliable, simple, robust and quantitative measurement of SARS-CoV-2 S-protein nAbs in blood, providing a powerful tool to accurately and closely monitor SARS-CoV-2 S-protein nAbs levels at a large population scale and offering more confidence to individuals and clinicians to fight this pandemic. In Phase I, the key focus is to prove the concept of rapid quantification of SARS-CoV-2 S-protein nAbs on the proposed testing device, and demonstrate the feasibility of the testing device in blood samples in terms of sensitivity, accuracy and detection range. The Pt-Pd MNFs enhanced smartphone platform is expected to have high accuracy (< 5% RSD) for SARS-CoV-2 S-protein nAbs quantification (1ng/ml – 100ug/ml). In phase II, the tests will be evaluated using clinic samples and further optimized for real-world applications. The proposed testing device, once validated, will greatly benefit public health in the U.S. as well as worldwide. There is a large market and significant commercial opportunity for such an effective testing device. 1