Antibody Binding Epitope Mapping (AbMap) of Hundred Antibodies in a Single Run

Abstract

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 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.

Advances and Utility of the Human Plasma Proteome

“…An alternative protein-based detection of circulating antibodies uses arrays of peptides, which can be produced in a short time using the viral genome sequence. − In the early COVID-19 pandemic, Yu et al developed a microarray with 966 15-mer peptides, which fully covers all of the SARS-CoV-2 proteins with which the landscape of B-cell epitopes for SARS-CoV-2 IgM and IgG antibodies in early COVID-19 infection was constructed . Shrock et al developed a VirScan phage-display platform containing an oligopeptide library across the proteomes of all known pathogenic human viruses (∼400 species and strains).…”

Development and characterization of a camelid derived antibody targeting a linear epitope in the hinge domain of human PCSK9 protein

“…However, the specific interaction epitope is yet unclear. To determine the interaction sites, we employed a random peptide library, phage display, and sequencing technology and characterized the amino acid of the binding epitope 17 , 18 .…”

Epitope Analysis of Anti-SARS-CoV-2 Neutralizing Antibodies

“…The AbMap technology proposed by Qi et al [49] enables high-throughput analysis of antibody-binding epitopes on antigens at amino acid-level resolution. AbMap has already been applied for dissecting SARS-CoV-2-specific epitopes [50] .…”