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

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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.