Development and Application of Antibody Discovery Technologies For Diverse Viral Pathogens

Abstract

Existing viral pathogens like human immunodeficiency virus (HIV), influenza virus, and coronaviruses, along with emerging infectious diseases, are constant worldwide threats, causing illness and death for millions of people each year. Studying the immune response to these pathogens, and specifically the antibody response, can guide the development of vaccines and therapeutic tools to prevent and treat these diseases. To efficiently and effectively screen B cells in order to characterize the antibody response, I helped develop an antibody discovery technology termed LIBRA-seq (Linking B cell receptor to antigen specificity through sequencing). LIBRA-seq allows for high throughput determination of antibody sequence and antigen specificity at the single cell level. First, I tested and validated the technology using cell lines with known B cell receptor sequences and specificities and demonstrated that LIBRA-seq could successfully discriminate diverse antigen reactivity patterns. Next, I used LIBRA-seq to identify broadly neutralizing HIV and influenza antibodies from HIV infection samples. Additionally, I characterized broadly reactive coronavirus antibodies from a SARS-CoV recovered donor. Further, I extended the LIBRA-seq technology to include a functional readout from sequencing data by including a viral protein and its receptor in the antigen screening library. I tested this idea with SARS-CoV-2 spike protein and the ACE2 receptor and showed high rates of neutralizing antibody discovery. These experiments highlighted the power and utility of LIBRA-seq while contributing discovery of novel antibodies to the field. Overall, the application of LIBRA-seq to diverse viral pathogens has led to analysis of antibody repertoires, and discovery of novel antibodies, enhancing our knowledge of antibody-antigen interactions and elucidating viral epitopes, all of which ultimately aid in vaccine design and therapeutic development.