Reagents to Chemically Tag Specific Coronavirus Spike Proteins

化学标记特定冠状病毒刺突蛋白的试剂

基本信息

批准号：
10259048
负责人：
STEVEN A BENNER
金额：
$ 25.89万
依托单位：
FIREBIRD BIOMOLECULAR SCIENCES, LLC
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-07 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10259048
关键词：
2019-nCoV Acylation Affinity Aminoacylation Antibodies Antigens Antiviral Agents Arboviruses Architecture Asia Binding Binding Proteins Biological Assay Biological Sciences Breast Cancer Cell COVID detection Cell surface Chemicals Chemistry Clinical Laboratory Information Systems Coronavirus Coronavirus Infections Coronavirus spike protein Crystallization Cystic Fibrosis DNA Detection Development Diagnostic Disulfides Electrodes Ensure Environment Europe Evolution Fright Generations Genes Image In Vitro India Individual Information Systems Institutional Review Boards Laboratories Lateral Lung diseases Lysine Malignant neoplasm of liver Measures Medical Middle East Respiratory Syndrome Molecular Conformation Norovirus Nucleic Acids Nucleotides Oxidation-Reduction Peptides Performance Phase Preparation Procedures Protein Engineering Proteins Publishing RNA Reaction Reagent Regulation Reproducibility Research Saliva Sampling Schools Science Severe Acute Respiratory Syndrome Specificity Structure Surface Testing Time Toxin Vaccines Virion Virus Work amino group analog antibody test aptamer base cancer cell catalyst covalent bond disulfide bond experience genetic information improved innovation instrument instrumentation novel coronavirus nuclease pandemic disease recruit respiratory pathogen response symptom treatment tool

项目摘要

Reagents to Chemically Tag Specific Coronavirus Spike Proteins Firebird Biomolecular Sciences, LLC. Steven A. Benner Bharat Gawande Abstract While vaccines and antivirals may be long term solutions to the current coronavirus pandemic, it is now widely appreciated that the pandemic can be managed before these emerge by rapid, point-of-sampling "public space entry tests" (PSETs), given appropriate FDA regulatory relief. PSETs will be even more important if a good vaccine never emerges, a possibility given experience with other coronaviruses. In the 2003 SARS crisis, we delivered a "best in class" PCR kit for SARS that targeted coronaviral RNA. This was possible due to our first generation nucleic acid innovations, which were also used in respiratory pathogen panels and cystic fibrosis tests, inter alia. Luminex acquired EraGen in 2011 for $34 million. Since then, the PI generated 2nd and 3rd generation innovations that supported tests for MERS, arboviruses, norovirus and, this year, CoV-19 itself. These include a PSET that gives 10 minute results using proprietary displaceable probe loop amplification. Intrinsic chemistry suggests, however, this is the fastest any RNA-targeted test can be. Here, we will use our 3rd-generation DNA innovations not to target coronaviral RNA, but the coronavirus itself. Firebird will create reagents (AEGISZymes) from an artificially expanded genetic information system (AEGIS) that catalyze covalent acylation of lysines on the surface of ~300 spike proteins per virus. Each acylation will generate ~300 redox active moieties on an electrode surface per virion. The test is conceptually advanced over antibody tests, as it uses stable covalent bond formation, rather than non-covalent antibody: antigen interactions, and exploits electrochemical detection rather than a lateral flow. Our Phase 2 partner, MightyGate, has shown with standard aptamers detection in less than a minute with such architectures. With >100x sensitivity over Abbott BinaxNOWTM, the assay should detect virus in any contagious individual at entrances to arenas, schools, and other spaces as fast has handbags and are checked. Specifically, we will: Aim 1. Under IRB 2020001, we will use AEGIS-LIVE to evolve AEGISZymes in saliva that acylate lysines in spike proteins and peptide loops of CoV-19, SARS, and MERS. Aim 2. Metric the AEGISZymes that emerge, measuring their binding affinities, acylation rates, and specificity among the 3 homologous surface loops. The metrics are: M1. Acylation with kcat ≈ 1 sec-1. This should be readily achieved based on pmolar affinities that are now routine with AEGIS-LIVE; similar rate constants are now routine in analogous catalysts. M2. Specificity with kcat/Kdiss ratios >103, to ensure that CoV-19 is distinguished from other coronaviruses. If the metrics are met, MightyGate has committed itself to participate in Phase 2 work that will incorporate AEGISZymes onto its kiosk-style instrument. The kiosk has been proven with standard aptamers to give electrochemical readouts in less than 1 minute. Firebird's AEGISZymes will immediately improve the sensitivity of that kiosk, and immediate commercial value. However, our approach is general to any virus target.

化学标记特异性冠状病毒峰值蛋白的试剂火鸟生物分子科学有限责任公司。史蒂文·本纳（Steven A. Benner）巴拉特·加瓦德（Bharat Gawande）抽象的虽然疫苗和抗病毒药可能是当前冠状病毒大流行的长期解决方案，但现在是广泛赞赏的是，大流行可以在这些大流行中通过快速的，抽样的点出现。鉴于适当的FDA调节缓解，太空进入测试”（PSET）。如果A 好疫苗从未出现过，这是有其他冠状病毒经验的可能性。在2003年的SARS危机中，我们为针对冠状病毒RNA的SARS提供了“最好的班级” PCR套件。这由于我们的第一代核酸创新，这也是可能的，这也用于呼吸病原体面板和囊性纤维化测试，除其他外。 Luminex在2011年以3400万美元的价格收购了Eragen。从那以后，Pi 产生了第二代和第三代创新，这些创新支持了MERS，Arboviruses，Norovirus的测试一年，COV-19本身。其中包括一个使用专有替代探针循环提供10分钟结果的PSET 放大。固有的化学表明，这是任何靶向RNA的测试最快的方法。在这里，我们将使用我们的第三代DNA创新来靶向冠状病毒RNA，而是冠状病毒本身。 Firebird将从人为扩展的遗传信息系统中创建试剂（私元）（宙斯盾）催化赖氨酸在每个病毒300个尖峰蛋白表面上的共价酰化。每个每个病毒体的电极表面上的酰化将产生约300个氧化还原活性部分。该测试在概念上是在抗体测试上进行了高级，因为它使用稳定的共价键形成，而不是非共价抗体：抗原相互作用，并利用电化学检测而不是横向流动。我们的第二阶段合作伙伴， Mightygate在不到一分钟的时间内就以标准的适体检测显示了此类体系结构。对Abbott Binaxnowtm的灵敏度> 100倍，该测定应检测到任何传染性个体的病毒竞技场，学校和其他空间的条目随着快速的方式提供手袋，并已进行检查。具体来说，我们将：目的1。在IRB 2020001下，我们将使用Aegis-live在唾液中进化，以在唾液中酰化赖氨酸的脂肪酶 COV-19，SARS和MERS的尖峰蛋白和胡椒环。 AIM 2。公制的销售的销售，测量其结合亲和力，酰化率和特异性在3个同源表面环中。指标是： M1。用KCAT≈1秒1酰基化。这应该根据现在的PMolar亲和力很容易实现宙斯盾的常规；类似的速率常数在类似催化剂中是常规的。 M2。具有KCAT/KDISS比率> 103的特异性，以确保COV-19与其他冠状病毒区分开。如果达到指标，Mightygate已承诺参加第二阶段的工作，该工作将纳入将其赋予其售货亭风格的仪器。售货亭已通过标准的适体证明不到1分钟的电化学读数。 Firebird的盟友将立即提高灵敏度该售货亭和直接的商业价值。但是，我们的方法是任何病毒靶标的一般方法。