Throughput, speed, resolution, and sample consumption are typically key limiting factors for performing detailed kinetic and epitope characterization of monoclonal antibody (mAb) libraries destined for use as therapeutics. In addition, mAb immune responses can be used to survey the epitope landscape of pathogens and inform the design of more effective immunogens for vaccines. Here, we demonstrate three core applications of high throughput SPR: capture kinetics, epitope binning, and epitope mapping, that together provide a comprehensive characterization of your mAb library with minimal sample consumption, enabling more confident decisions to be made earlier in the research process and obviating the need for preliminary ELISA screening. We show that high throughput SPR can facilitate the generation of high-quality kinetic data from 384 mAbs in parallel, using < 1 μg per mAb and only 2 μg of antigen. Epitope binning in a 384-mAb array format allows for a rapid assessment of the depth and breadth of your library’s epitope diversity, providing exquisite resolution between near-identical clones, using < 5 μg per mAb and about 100 μg antigen. Finally, a large panel of mAbs can be mapped against a 384-peptide library, if their epitopes can be (partially) recapitulated on peptides, using 1 μg per mAb and <0.1 μg per peptide.