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 intellectual property. Given the millions of antibodies for which epitope information is lacking, 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. Using AbMap, profiles of the peptides bound by 202 antibodies were determined in a single test, and linear epitopes were identified for >50% of the antibodies. Using spike protein (S1 and S2)-enriched antibodies from the convalescent serum of one COVID-19 patient as the input, both linear and potentially conformational epitopes of spike protein specific antibodies were identified. We defined peptide-binding profile of an antibody as the binding capacity (BiC). Conceptually, the BiC could serve as a systematic and functional descriptor of any antibody. Requiring at least one order of magnitude less time and money to map linear epitopes than traditional technologies, AbMap allows for high-throughput epitope mapping and creates many possibilities. [Display omitted] •The first technology enables epitope mapping of 200 antibodies in a single run.•Applications of antibodies, not for original antigens, were further exploited.•The epitopes of antibodies in one COVID-19 patient were identified.•The BiC is a new systematic and functional character of antibody. In this study, we developed a strategy, Antibody binding epitope Mapping (AbMap), by combining a phage displayed peptide library with next-generation sequencing. By AbMap, epitopes of 202 antibodies were determined in a single test. Also, the epitopes of spike protein-specific antibodies from convalescent serum of one COVID-19 patient 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.