Abstract
Prevention of Epstein-Barr virus (EBV) infection has focused on generating neutralizing antibodies (nAbs) targeting the major envelope glycoprotein gp350/220 (gp350). In this study, we generated 23 hybridomas producing gp350-specific antibodies. We compared the candidate gp350-specific antibodies to the well-characterized nAb 72A1 by: (1) testing their ability to detect gp350 using enzyme-linked immunosorbent assay, flow cytometry, and immunoblot; (2) sequencing their heavy and light chain complementarity-determining regions (CDRs); (3) measuring the ability of each monoclonal antibody (mAb) to neutralize EBV infection in vitro; and (4) mapping the gp350 amino acids bound by the mAbs using competitive cell and linear peptide binding assays. We performed sequence analysis to identify 15 mAbs with CDR regions unique from those of murine 72A1 (m72A1). We observed antigen binding competition between biotinylated m72A1, serially diluted unlabeled gp350 nAbs (HB1, HB5, HB11, HB20), and our recently humanized 72A1, but not gp350 non-nAb (HB17) or anti-KSHV gH/gL antibody.
Keywords:
Complementarity-determining region; Epitope; Epstein-Barr virus; Immunodominant; Immunosuppression; Infection; Neutralizing antibodies; gp350.
Copyright © 2019 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / isolation & purification
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Antibodies, Monoclonal / pharmacology
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Antibodies, Neutralizing / biosynthesis
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Antibodies, Neutralizing / chemistry*
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Antibodies, Neutralizing / isolation & purification
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Antibodies, Neutralizing / pharmacology
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Antibodies, Viral / biosynthesis
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Antibodies, Viral / chemistry*
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Antibodies, Viral / isolation & purification
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Antibodies, Viral / pharmacology
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B-Lymphocytes / immunology
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B-Lymphocytes / virology
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Binding Sites, Antibody
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Binding, Competitive
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Cell Line, Tumor
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Complementarity Determining Regions / chemistry
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Complementarity Determining Regions / immunology
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Enzyme-Linked Immunosorbent Assay
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Epithelial Cells / immunology
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Epithelial Cells / virology
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Epstein-Barr Virus Infections / immunology
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Epstein-Barr Virus Infections / prevention & control
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Epstein-Barr Virus Infections / virology
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Herpesvirus 4, Human / drug effects*
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Herpesvirus 4, Human / genetics
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Herpesvirus 4, Human / immunology
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Humans
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Hybridomas / chemistry
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Hybridomas / immunology
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Immunodominant Epitopes / chemistry*
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Immunodominant Epitopes / immunology
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Mice
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Protein Binding
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Sequence Alignment
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Sequence Homology, Amino Acid
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Viral Matrix Proteins / chemistry*
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Viral Matrix Proteins / immunology
Substances
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Antibodies, Monoclonal
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Antibodies, Neutralizing
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Antibodies, Viral
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Complementarity Determining Regions
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EBV-associated membrane antigen, Epstein-Barr virus
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Immunodominant Epitopes
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Viral Matrix Proteins