1.Identification of Dengue-specific B-Cell Epitopes by Phage-display Random Peptide Library
Nevis Amin ; Alicia Aguilar ; Frank Chamac ho ; Yaime Vázquez ; Maritza Pupo ; Juan Carlos Ramirez ; Luis Izquierdo ; Felix Dafhnis ; David Ian Stott ; Ela Maria Perez ; Armando Acosta
Malaysian Journal of Medical Sciences 2009;16(4):4-14
Background: Dengue is the most important human viral disease transmitted by arthropod
vectors. The availability of random peptide libraries (RPL) displayed on phage has provided a powerful
tool for selecting sequences that mimic epitopes from microorganisms that are useful for diagnostic
and vaccine development purposes. In this paper, we describe peptides that resemble the antigenic
structure of B-cell epitopes of dengue virus identified from a phage-peptide library using human sera
containing polyclonal antibodies against dengue virus.
Materials and Methods: Eighteen phage clones were isolated from the phage-display peptide
library, J404, by affinity selection using human antisera against dengue virus type 3. These clones were
tested for reactivity by ELISA with a panel of hyperimmune ascitic fluids (HAFs) containing antibodies
either against all four dengue serotypes, West Nile virus (WNV) or Eastern equine encephalitis virus
(EEEV) with control ascitic fluid (NAF) used as a negative control.
Results: Eight clones were recognized by HAFs against the four dengue serotypes, of which
four significantly inhibited binding of anti-dengue antibodies to the virus. Two peptides with similar
sequences to regions of NS3 and NS4B non-structural dengue virus proteins were identified.
Conclusion: Our results suggest that these peptides could be used for the development of
diagnostic tools for the detection of dengue virus infection and for a potential vaccine against this
pathogen.
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