Nanobody toolkit for human coronavirus classification

用于人类冠状病毒分类的纳米抗体工具包

基本信息

批准号：
10375561
负责人：
ANDREW HAYHURST
金额：
$ 24.75万
依托单位：
TEXAS BIOMEDICAL RESEARCH INSTITUTE
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：
2021-03-19 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10375561
关键词：
2019-nCoV Affinity Amino Acid Sequence Amino Acids Animals Antibodies Antibody Response Antigen Targeting Appearance Architecture Attention Binding Biological Assay Biological Markers Cells Classification Common Cold Communities Coronavirus Coronavirus Infections Coronavirus nucleocapsid protein Deposition Diagnostic Discrimination Disease Ecosystem Elements Ensure Enzyme-Linked Immunosorbent Assay Enzymes Epidemic Epitopes Escherichia coli Family Funding Future Genbank Genes Genome Goals Haptens Health Human Hybridomas Immune Sera Immunization Immunoglobulin G Individual Libraries Llama Logic Mediating Microscopy Middle East Respiratory Syndrome Middle East Respiratory Syndrome Coronavirus Minor Molecular Conformation Nucleocapsid Nucleocapsid Proteins Nucleoproteins Population Proteins RNA Reagent Recombinants Reporter Research Research Personnel Research Proposals Respiratory Disease Respiratory distress SARS coronavirus Sequence Homology Serum Severe Acute Respiratory Syndrome Specificity Structure System Technology Therapeutic Intervention United States National Institutes of Health Variant Virus Western Blotting Work Zoonoses antigen test base cross reactivity dimer follow-up global health human coronavirus human disease human pathogen in silico innovation milligram nanobodies novel novel coronavirus pandemic disease particle pathogen respiratory response screening seroconversion spillover event success zoonotic coronavirus zoonotic spillover

项目摘要

ABSTRACT The endemic human coronaviruses (HCoV) NL63, OC43, 229E and HKU1 have typically been associated with relatively mild and self-limiting respiratory disease resembling the common cold in healthy individuals. However, the newly emerging zoonotic coronaviruses (SARS-CoV, MERS-CoV and SARS-CoV-2) can cause more severe respiratory distress, especially in those with underlying health conditions and can be fatal. Coronaviruses are diverse and spillover from zoonotic reservoirs either directly to humans or via an intermediate host. Spillover is likely to be more frequent due to the ever-expanding human population and ecosystem erosion, thereby ensuring more collisions between natural and unnatural host. Our exploratory research proposal seeks to develop a small toolkit of immunoreagents that will form the basis of a logic gate to identify and classify coronaviruses. A single immunoreagent, with broad recognition of overall shared protein architecture, will allow the identification of the coronavirus genera. Separate immunoreagents with absolute specificity for unique amino-acid variation between the human and zoonotic coronaviruses will allow the further classification to a known pathogen. Such a toolkit will quickly establish if a newly emerging virus is a coronavirus we are familiar with or is a novel genus or lineage that has never before spilled over into the human population and may demand urgent follow up for epidemic or pandemic potential. Our immunoreagents are based on llama single domain antibodies or nanobodies that are inexpensive to produce at high yields in E. coli and highly modular, allowing fusion to enzymatic reporter enzymes for one-step probing of virus infected cells and facile establishment of sandwich assays. Immortalization of nanobody sequences in silico means these are immediately available to the wider research community following deposition in Genbank unlike polyclonal sera or hybridomas secreting IgG. Single-pot library technology enables a rapid response to be mounted against any newly emerged coronavirus in the future, ensuring we are more pro-active than re-active to help safeguard human health.

抽象的地方性人类冠状病毒 (HCoV) NL63、OC43、229E 和 HKU1 通常被与相对轻微且自限性的呼吸道疾病有关，类似于健康人的普通感冒个人。然而，新出现的人畜共患冠状病毒（SARS-CoV、MERS-CoV 和 SARS-CoV-2）可能会导致更严重的呼吸窘迫，尤其是那些有潜在健康问题的人，并且可能会致命的。冠状病毒多种多样，可以从人畜共患病宿主直接传播到人类，也可以通过传播途径传播给人类。中间宿主。由于人口不断增长，溢出效应可能会更加频繁生态系统侵蚀，从而确保自然和非自然宿主之间更多的碰撞。我们的探索研究提案旨在开发一个小型免疫试剂工具包，该工具包将构成逻辑门的基础识别和分类冠状病毒。单一免疫试剂，可广泛识别整体共享蛋白质架构，将允许识别冠状病毒属。绝对分离免疫试剂人类和人畜共患冠状病毒之间独特氨基酸变异的特异性将允许进一步研究对已知病原体进行分类。这样的工具包将快速确定新出现的病毒是否是我们熟悉的冠状病毒或者是一种以前从未扩散到全球的新属或谱系人口，可能需要紧急跟进流行病或大流行的可能性。我们的免疫试剂基于美洲驼单域抗体或纳米抗体，这些抗体或纳米抗体可以在大肠杆菌中以低廉的成本高产量生产。大肠杆菌和高度模块化，允许与酶报告酶融合，对感染的病毒进行一步探测细胞并轻松建立夹心测定法。纳米抗体序列的计算机永生化与 Genbank 不同的是，这些内容在存放于 Genbank 后可立即提供给更广泛的研究界分泌 IgG 的多克隆血清或杂交瘤。单罐文库技术可实现快速响应应对未来任何新出现的冠状病毒，确保我们更加主动而不是被动帮助维护人类健康。