Development of a High-Throughput Microfludics-Enabled Functional Assay for Rapidly Identifying Neutralizing Antibodies
We will generate stocks of influenza pandemic A/California/04/09 H1N1 (pH1N1) and A/Wyoming/3/03 H3N2 influenza A viruses as well as influenza B/Brisbane/60/08 mCherry-expressing viruses that will be used to optimize the performance of PRESCIENT (Aim 1), to test the working hypothesis that PRESCIENT can identify in a mixed population of individual human hybridoma cells neutralizing antibodies against pH1N1 virus (Aim 2) and to identify influenza A H1N1, H3N2; and B virus neutralizing antibodies from EBV-immortalized patient-derived B cells (Aim 3). If needed, we will also generate stocks of the single-cycle infectious influenza pH1N1 viruses pseudotyped with the HA of influenza A pH1N1, H3N2; and B viruses (Aim 1). We will also perform traditional neutralization assays for the confirmation of the positive hits identified in the PRESCINT screening with patient-derived B cells (Aim 3).