title: Potent dengue virus neutralization by a therapeutic antibody with low monovalent affinity requires bivalent engagement.
abstract: We recently described our most potently neutralizing monoclonal antibody, E106, which protected against lethal Dengue virus type 1 (DENV-1) infection in mice. To further understand its functional properties, we determined the crystal structure of E106 Fab in complex with domain III (DIII) of DENV-1 envelope (E) protein to 2.45 Å resolution. Analysis of the complex revealed a small antibody-antigen interface with the epitope on DIII composed of nine residues along the lateral ridge and A-strand regions. Despite strong virus neutralizing activity of E106 IgG at picomolar concentrations, E106 Fab exhibited a ∼20,000-fold decrease in virus neutralization and bound isolated DIII, E, or viral particles with only a micromolar monovalent affinity. In comparison, E106 IgG bound DENV-1 virions with nanomolar avidity. The E106 epitope appears readily accessible on virions, as neutralization was largely temperature-independent. Collectively, our data suggest that E106 neutralizes DENV-1 infection through bivalent engagement of adjacent DIII subunits on a single virion. The isolation of anti-flavivirus antibodies that require bivalent binding to inhibit infection efficiently may be a rare event due to the unique icosahedral arrangement of envelope proteins on the virion surface.
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Edeling
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: S Kyle
authors.initials: SK
authors.lastname: Austin
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Bimmi
authors.initials: B
authors.lastname: Shrestha
authors.affiliation: Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Dowd
authors.affiliation: Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Swati
authors.initials: S
authors.lastname: Mukherjee
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Nelson
authors.affiliation: MacroGenics, Rockville, Maryland, United States of America.
authors.forename: Syd
authors.initials: S
authors.lastname: Johnson
authors.affiliation: Integral Molecular, Philadelphia, Pennsylvania, United States of America.
authors.forename: Manu N
authors.initials: MN
authors.lastname: Mabila
authors.affiliation: Integral Molecular, Philadelphia, Pennsylvania, United States of America.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Christian
authors.affiliation: Integral Molecular, Philadelphia, Pennsylvania, United States of America.
authors.forename: Joseph
authors.initials: J
authors.lastname: Rucker
authors.affiliation: Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Theodore C
authors.initials: TC
authors.lastname: Pierson
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, United States of America; Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Diamond
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America; Department of Biochemistry & Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Daved H
authors.initials: DH
authors.lastname: Fremont
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D000915:Antibody Affinity; D003715:Dengue / Q000188:drug therapy / Q000276:immunology; D003716:Dengue Virus / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D051379:Mice; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014771:Virion / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000939:Epitopes; D007074:Immunoglobulin G; D014759:Viral Envelope Proteins; C072281:E protein TH Sman, Dengue virus
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