摘要：SummaryUnderstanding the antibody response is critical to developing vaccine and antibody-based therapies and has inspired the recent development of new methods to isolate antibodies. Methods to define the antibody-antigen interactions that determine specificity or allow escape have not kept pace. We developed Phage-DMS, a method that combines two powerful approaches—immunoprecipitation of phage peptide libraries and deep mutational scanning (DMS)—to enable high-throughput fine mapping of antibody epitopes. As an example, we designed sequences encoding all possible amino acid variants of HIV Envelope to create phage libraries. Using Phage-DMS, we identified sites of escape predicted using other approaches for four well-characterized HIV monoclonal antibodies with known linear epitopes. In some cases, the results of Phage-DMS refined the epitope beyond what was determined in previous studies. This method has the potential to rapidly and comprehensively screen many antibodies in a single experiment to define sites essential for binding interactions.Graphical AbstractDisplay OmittedHighlights•A high-throughput, comprehensive method to identify antibody epitopes is needed•Phage-DMS combines phage display technology and deep mutational scanning•Phage-DMS identified single mutations that lead to escape from HIV Env antibody binding•Effect of mutations in Phage-DMS correlate with results using a parallel approachVirology; Genomic Library
