Antibody binding epitope Mapping (AbMap) of hundred antibodies in a single run

Antibodies play essential roles in both diagnostics and therapeutics. Epitope mapping is essential to understand how an antibody works and to protect the intellectual property. Given millions of antibodies without epitope information, there is a need for high-throughput epitope mapping. To address this, we developed a strategy, Antibody binding epitope Mapping (AbMap), by combining a phage displayed peptide library with next generation sequencing. By AbMap, profiles of the binding peptides of 202 antibodies were determined in a single test, and linear epitopes were identified for >50% of the antibodies. Using the spike protein (S1 and S2) enriched antibodies from convalescent serum of one COVID-19 patient as input, both linear and conformational epitopes of spike protein specific antibodies were identified. We defined the profile of the binding peptides of an antibody as Binding Capacity (BiC), conceptually, BiC could serve as a systematic and functional character for any antibody. By mapping linear epitope at least one order of magnitude lower of time and cost than that of traditional technologies, AbMap enables high-throughput epitope mapping and opens many possibilities.