Identification of a dengue virus type 2 (DEN-2) serotype-specific B-cell epitope and detection of DEN-2-immunized animal serum samples using an epitope-based peptide antigen

Abstract

In this study, a serotype-specific monoclonal antibody (mAb), D 2 16-1 (Ab4), against dengue virus type 2 (DEN-2) was generated. The specificity of Ab4, which recognized DEN-2 non-structural protein 1, was determined by ELISA, immunofluorescence and immunoblotting analyses. The serotype-specific B-cell epitope of Ab4 was identified further from a random phage-displayed peptide library; selected phage clones reacted specifically with Ab4 and did not react with other mAbs. Immunopositive phage clones displayed a consensus motif, His-Arg/Lys-Leu/Ile, and a synthetic peptide corresponding to the phage-displayed peptide bound specifically to Ab4. The His and Arg residues in this epitope were found to be crucial for peptide binding to Ab4 and binding activity decreased dramatically when these residues were changed to Leu. The epitope-based synthetic peptide not only identified serum samples from DEN-2-immunized mice and rabbits by ELISA but also differentiated clearly between serum samples from DEN-2-and Japanese encephalitis virus-immunized mice. This mAb and its epitope-based peptide antigen will be useful for serologic diagnosis of DEN-2 infection. Furthermore, DEN-2 epitope identification makes it feasible to dissect antibody responses to DEN and to address the role of antibodies in the pathogenesis of primary and secondary DEN-2 infections.

A Novel Peptide Specifically Binding to Nasopharyngeal Carcinoma For Targeted Drug Delivery

“…Therefore, the development of additional targeted technologies is highly desirable. Recently, phage-displayed peptide libraries have been used to select peptides that bind to specific receptors (12,13) or antibodies (14,15). Strategies for panning cells in vitro (16,17) or tissues in vivo (18 -22) with complex phage libraries have been described to yield phages with organ-or tumor-binding specificity.…”

Recombinant Multiepitope Protein for Early Detection of Dengue Infections

“…One way to address these concerns is to replace the whole virus antigens with customized recombinant antigens, consisting of carefully chosen epitopes. Immunodominant, linear dengue viral epitopes, potentially suitable for constructing multiepitope proteins, have been mapped extensively on the major structural protein, the envelope (E) protein (12,15,20), and the nonstructural (NS) protein NS1 (3,4,22,23). The majority of these epitopes specifically recognize IgG antibodies.…”

Generation and Characterization of Monoclonal Antibodies against Dengue Virus Type 1 for Epitope Mapping and Serological Detection by Epitope-Based Peptide Antigens

“…A phage-displayed 12-mer peptide library was purchased from New England Biolabs, Inc. (Beverly, MA). Phage display biopanning procedures were performed according to our previously published method (61). The titer of the unamplified third-round phage particles was determined on Luria-Bertani medium-IPTG (isopropyl-␤-D-thiogalactopyranoside)-X-Gal (5-bromo-4-chloro-3-indolyl-␤-D-galactopyranoside) plates, and immunopositive phage clones were screened by ELISA.…”

“…Phage-displayed random peptide libraries provide opportunities to map B-cell epitopes (11,14,52,60,61) and protein-protein contacts (3,7,42,53), select bioactive peptides bound to receptors (26,33) or proteins (7,9,10,27,44), search for disease-specific antigen mimics (13,36,46), and determine cell-specific (4, 31, 39) and organ-specific (2,12,45) peptides.…”