Use of recombinant fusion proteins and monoclonal antibodies to define linear and discontinuous antigenic sites on the dengue virus envelope glycoprotein

Virology. 1992 Apr;187(2):480-91. doi: 10.1016/0042-6822(92)90450-4.


Sixteen overlapping fragments of the dengue-2 virus envelope (E) protein, expressed as trpE-E fusion products in Escherichia coli, were used to map the epitopes defined by a panel of 20 monoclonal antibodies (MAbs) by immunoblotting. Using this technique, the amino acid sequence of six antigenic domains on the E protein was characterized. Nonneutralizing MAbs were found to define either linear-specific, subcomplex-specific (amino acids 22-58), and complex-specific (amino acids 304-332) epitopes or a subcomplex conformational-dependent epitope requiring the presence of two closely linked amino acid sequences from the E protein, 60-97 and 298-397. Neutralizing MAbs, however, defined either group-reactive epitopes present on two overlapping domains (amino acids 60-135; amino acids 60-205) or type-, subcomplex-, complex-, subgroup-, and group-specific determinants (amino acids 298-397). These neutralizing epitopes were all found to be dependent upon disulfide bridges. Our results suggest that the maintenance of a topographical arrangement of discontinuous antigenic domains in the flavivirus E-protein is necessary to induce neutralizing and protective antibodies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Viral / immunology
  • Antigens, Viral / chemistry
  • Antigens, Viral / immunology*
  • Cloning, Molecular
  • Dengue Virus / immunology*
  • Disulfides
  • Epitopes
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / immunology
  • Molecular Sequence Data
  • Recombinant Fusion Proteins / immunology
  • Viral Envelope Proteins / chemistry
  • Viral Envelope Proteins / immunology*


  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • Disulfides
  • Epitopes
  • Membrane Glycoproteins
  • Recombinant Fusion Proteins
  • Viral Envelope Proteins