Antibody affinity is a critical metric for antibody therapeutic development and is often only assessed after initial antibody discovery hit generation. However, due to the cost and throughput of small-scale antibody production, antibody affinity characterization is only reserved for a few lead candidates. Here, we demonstrate a high-throughput relative affinity ranking of recombinant antigen-specific antibodies during the antibody discovery workflow on Bruker’s Beacon optofluidic system.