RAPID: Large-scale functional analysis of antibody repertoires elicited by SARS-CoV-2

RAPID：对 SARS-CoV-2 引发的抗体库进行大规模功能分析

• 批准号: 2029949
• 负责人: Paul de Figueiredo
• 金额: $ 20万
• 依托单位: Texas A&M University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2029949&HistoricalAwards=false
• 关键词: RAPID; Large; scale; functional; analysis

The award to Texas A&M University will support research on a novel lab-on-a-chip system, to determine the identity, functionality, and clonality of antibodies (Abs) elicited by SARS-CoV-2, the causative agent of COVID-19. One of the most important functions of Abs during viral infection is neutralization, the process whereby Ab binding to viral or host targets prevents viral entry into host cells, thereby thwarting disease. Conventional approaches for determining the functions of pathogen-specific neutralizing Ab (nAb) repertoire are time- and labor-intensive, and thus have only been used to investigate small portions of Ab repertoires. This award will support research to develop a system that enables fast, direct, functional assays that measure viral neutralization activities of Ab-producing cells. In addition to the strong scientific impact, this award will support cross-disciplinary training postdoctoral, graduate student, and undergraduate student trainees, including women and scientists from historically under-represented groups. Results from the studies will be disseminated rapidly and in peer-reviewed journals and at scientific meetings.Until recently, the large-scale analysis of antibody repertoires was cost-prohibitive, owing to their massive sizes. However, over the past decade, novel sequencing approaches, including Ig-seq, have provided unprecedented insight into Ab gene repertoires. However, despite these dramatic advances, our understanding of the functions of Ab repertoires remains incomplete. Support from this award will advance a droplet microfluidics platform termed PRESCIENT, that enables fast, single-cell (digital) resolution, direct, functional assays that measure viral neutralization activities of Ab-producing B-cells, thereby providing a system for rapidly characterizing a large repertoire of nAbs against viral pathogens. The overall objectives are to deliver: A fully integrated microfluidic platform with dual fluorescence detection capability and system level throughput of at least 10 assays/sec achieved; demonstration that sera from inoculated mice react with recombinant SARS-CoV-2 spike protein antigen; and characterization (at single cell resolution) the nAb (functional) repertoire against pseudotyped SARS-CoV-2-GFP. This RAPID award will thus deliver the first global functional characterization of Abs that neutralize pseudotyped SARS-CoV-2, a timely response to the ongoing global pandemic caused by SARS-CoV-2 and an important complement to the ongoing computer simulation studies conducted by other labs. This RAPID award is made by the Division of Biological Infrastructure (DBI) using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

授予德克萨斯A M大学的奖项将支持对新型芯片实验室系统的研究，以确定由SARS-CoV-2（COVID-19的病原体）引发的抗体（Abs）的身份，功能和克隆性。在病毒感染期间，Ab的最重要功能之一是中和，即Ab与病毒或宿主靶标结合阻止病毒进入宿主细胞，从而阻止疾病的过程。用于确定病原体特异性中和Ab（nAb）库的功能的常规方法是时间和劳动密集型的，因此仅用于研究小部分Ab库。该奖项将支持研究开发一种系统，该系统能够快速，直接，功能性测定，测量Ab产生细胞的病毒中和活性。 除了强大的科学影响力外，该奖项还将支持跨学科培训博士后，研究生和本科生学员，包括来自历史上代表性不足的群体的女性和科学家。 研究结果将迅速在同行评审的期刊和科学会议上传播，直到最近，由于抗体库规模庞大，大规模分析成本高昂。然而，在过去的十年中，包括Ig-seq在内的新型测序方法为Ab基因库提供了前所未有的见解。然而，尽管这些戏剧性的进步，我们的理解的功能抗体剧目仍然是不完整的。 该奖项的支持将推进一个名为PRESCIENT的液滴微流体平台，该平台能够实现快速，单细胞（数字）分辨率，直接，功能性测定，测量产生Ab的B细胞的病毒中和活性，从而提供一个快速表征针对病毒病原体的大量nAb的系统。总体目标是提供：一个完全集成的微流控平台，具有双重荧光检测能力和系统水平的吞吐量至少达到10次测定/秒;证明接种小鼠的血清与重组SARS-CoV-2刺突蛋白抗原反应;和表征（在单细胞分辨率）针对假型SARS-CoV-2-GFP的nAb（功能）库。因此，RAPID奖将首次提供全球功能特性的抗体，中和假型SARS-CoV-2，及时应对正在进行的全球大流行引起的SARS-CoV-2和正在进行的计算机模拟研究的重要补充其他实验室。该奖项由生物基础设施部（DBI）使用冠状病毒援助，救济和经济安全（CARES）法案的资金颁发。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。