Identification of Mimotopes of the Japanese Encephalitis Virus Envelope Protein using Phage-Displayed Combinatorial Peptide Library
Cheng‐Wen Lin, Suh‐Chin Wu
Abstract
Cheng‐Wen Lin, Suh‐Chin Wu
Abstract
The phage-displayed combinatorial peptide library is a revolutionary method for discovering epitopes, in particular conformational epitopes. In this study, we characterized a Japanese encephalitis virus (JEV) conformational epitope by biopanning of phage-displayed random peptide libraries with a JEV envelope (E) protein-specific monoclonal antibody (mAb) 2H2. Eleven identified phage clones with high affinity to mAb 2H2 were identified using direct and inhibitory binding ELISA. Sequence alignment, structure modeling and mutational analysis revealed that the identified mimotopes for mAb 2H2 possess a conserved motif X(1)(D/E)(Y/T/S)X(2), fitting into a region at the domain III lateral surface of the E protein. The results of our study could provide useful information on the development of effective mimotope-based vaccines and diagnostic kits for the JEV infection.
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The phage-displayed combinatorial peptide library is a revolutionary method for discovering epitopes, in particular conformational epitopes. In this study, we characterized a Japanese encephalitis virus (JEV) conformational epitope by biopanning of phage-displayed random peptide libraries with a JEV envelope (E) protein-specific monoclonal antibody (mAb) 2H2. Eleven identified phage clones with high affinity to mAb 2H2 were identified using direct and inhibitory binding ELISA. Sequence alignment, structure modeling and mutational analysis revealed that the identified mimotopes for mAb 2H2 possess a conserved motif X(1)(D/E)(Y/T/S)X(2), fitting into a region at the domain III lateral surface of the E protein. The results of our study could provide useful information on the development of effective mimotope-based vaccines and diagnostic kits for the JEV infection.
Key concepts: Mimotope, Biopanning, Epitope, Phage display, Peptide library, Monoclonal antibody, Virology, Linear epitope