title: Complete mapping of viral escape from neutralizing antibodies.
abstract: Identifying viral mutations that confer escape from antibodies is crucial for understanding the interplay between immunity and viral evolution. We describe a high-throughput approach to quantify the selection that monoclonal antibodies exert on all single amino-acid mutations to a viral protein. This approach, mutational antigenic profiling, involves creating all replication-competent protein variants of a virus, selecting with antibody, and using deep sequencing to identify enriched mutations. We use mutational antigenic profiling to comprehensively identify mutations that enable influenza virus to escape four monoclonal antibodies targeting hemagglutinin, and validate key findings with neutralization assays. We find remarkable mutation-level idiosyncrasy in antibody escape: for instance, at a single residue targeted by two antibodies, some mutations escape both antibodies while other mutations escape only one or the other. Because mutational antigenic profiling rapidly maps all mutations selected by an antibody, it is useful for elucidating immune specificities and interpreting the antigenic consequences of viral genetic variation.
authors.affiliation: Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.|Department of Genome Sciences, University of Washington, Seattle, Washington, United States of America.|Medical Scientist Training Program, University of Washington, Seattle, Washington, United States of America.
authors.forename: Michael B
authors.identifier: 0000-0002-8172-6342
authors.initials: MB
authors.lastname: Doud
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Hensley
authors.affiliation: Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.|Department of Genome Sciences, University of Washington, Seattle, Washington, United States of America.
authors.forename: Jesse D
authors.identifier: 0000-0003-1267-3408
authors.initials: JD
authors.lastname: Bloom
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D014644:Genetic Variation; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics* / Q000276:immunology; D059014:High-Throughput Nucleotide Sequencing; D057131:Immune Evasion / Q000235:genetics* / Q000276:immunology; D009154:Mutation; D009500:Neutralization Tests; D009975:Orthomyxoviridae / Q000235:genetics* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus