Mimotope identification of human neutralizing monoclonal antibodies against dengue virus using random peptide phage display library

Abstract

Background: Dengue virus is the fastest emerging arboviral infection spread by Aedes mosquitoes with major public health consequences in over 100 tropical and sub-tropical regions. The DENV complex consists of four distinct but related viruses, designated as serotypes. Currently, there is no specific antiviral or vaccine is available for dengue virus infection. However, many researchers have been trying to develop the therapeutic agents against DENV. Phage display technology is a powerful method recently developed to screen selective repertories from large diversity of combinatorial peptide libraries displayed on the surface of bacteriophage. Methods: Two Human monoclonal antibodies (huMAbs) that show cross-neutralizing activity to 4 serotypes of DENV were produced from previous study (Setthapramote et al., 2012) and used as target antibodies for epitope study. Random 7 and 12 position peptide libraries were used for selection by 3 rounds of panning, and checked the target-specific phage clones by ELISA, before sending the M13 phage plasmids for sequencing. Results: The MAbs 19 and 54 recognized phages displaying peptides with the consensus motif LXXXG and LXGXG, respectively, that matched to conserved region of dengue E protein at position (107) LFGKG(111). However, for further specificity confirmation of the peptide, it need to be synthesized and checked the specificity by ELISA. The results described in this study will open avenues of development for dengue diagnostic assays and the technique can be applied for other antibodies specific to other tropical diseases.