Discontinuous Epitopes of Hepatitis B Surface Antigen Derived from a Filamentous Phage Peptide Library

The structure of the small hepatitis B virus surface antigen (HBsAg) was investigated by epitope mapping of four anti-HBsAg monoclonal antibodies (mAbs). Amino acid sequences of epitopes were derived from affinity-enrichment experiments (biopanning) using a filamentous phage peptide library. The lib...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1996-03, Vol.93 (5), p.1997-2001
Main Authors: Y.-C. Jack Chen, Delbrook, Kathy, Dealwis, Chris, Mimms, Larry, Mushahwar, Isa K., Mandecki, Woldek
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Antibodies
Antibodies, Monoclonal - immunology
Antigens
Bacteriophages
Binding, Competitive
Coliphages
Disulfides
Epitope Mapping
Epitopes
Gene Library
Hepatitis
Hepatitis antigens
Hepatitis B
Hepatitis B Antibodies - immunology
Hepatitis B Surface Antigens - immunology
Hepatitis B virus
Hepatitis B virus - immunology
Immunity (Disease)
Libraries
Molecular biology
Molecular Sequence Data
Protein Structure, Tertiary
Proteins
Surface antigens
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Summary:The structure of the small hepatitis B virus surface antigen (HBsAg) was investigated by epitope mapping of four anti-HBsAg monoclonal antibodies (mAbs). Amino acid sequences of epitopes were derived from affinity-enrichment experiments (biopanning) using a filamentous phage peptide library. The library consists of 109 different clones bearing a 30-residue peptide fused to gene III. Sequence homologies between peptides obtained from panning the library against the antibodies and the native HBsAg sequence allowed for precise description of the binding regions. Three of four mAbs were found to bind to distinct discontinuous epitopes between amino acid residues 101 and 207 of HBsAg. The fourth mAb was demonstrated to bind to residues 121-124. The sequence data are supported by ELISA assays demonstrating the binding of the HBsAg-specific peptides on filamentous phage to mAbs. The sequence data were used to map the surface of HBsAg and to derive a topological model for the α -carbon trace of the 101-207 region of HBsAg. The approach should be useful for other proteins for which the crystal structure is not available but a representative set of mAbs can be obtained.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.93.5.1997