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

该奖项授予德州农工大学（Texas a&m University），将支持一种新型芯片实验室系统的研究，以确定由COVID-19病原体SARS-CoV-2引发的抗体（Abs）的身份、功能和克隆性。抗体在病毒感染过程中最重要的功能之一是中和，即抗体与病毒或宿主靶标结合的过程，阻止病毒进入宿主细胞，从而阻止疾病。确定病原体特异性中和Ab （nAb）库功能的传统方法耗时耗力，因此仅用于研究一小部分Ab库。该奖项将支持开发一种系统的研究，该系统能够快速、直接、功能性地测量产生抗体的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群体的女性和科学家。这些研究的结果将在同行评议的期刊和科学会议上迅速传播。直到最近，由于抗体库的庞大规模，对其进行大规模分析的成本过高。然而，在过去的十年中，新的测序方法，包括Ig-seq，提供了前所未有的对Ab基因库的了解。然而，尽管有这些戏剧性的进展，我们对Ab库功能的理解仍然不完整。该奖项的支持将推进一种名为PRESCIENT的微流体液滴平台，该平台能够实现快速、单细胞（数字）分辨率、直接、功能分析，测量产生抗体的b细胞的病毒中和活性，从而提供一种快速表征大量抗病毒病原体的nab的系统。总体目标是提供：一个完全集成的微流控平台，具有双荧光检测能力和至少10次/秒的系统级吞吐量；证明接种小鼠血清与重组SARS-CoV-2刺突蛋白抗原发生反应；并在单细胞分辨率下鉴定抗伪型SARS-CoV-2-GFP的nAb（功能）库。因此，该RAPID奖将首次对中和假型SARS-CoV-2的抗体进行全球功能表征，及时应对SARS-CoV-2引起的持续全球大流行，并对其他实验室正在进行的计算机模拟研究进行重要补充。该RAPID奖由生物基础设施部（DBI）使用《冠状病毒援助、救济和经济安全（CARES）法案》的资金颁发。该奖项反映了美国国家科学基金会的法定使命，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。

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.