Fine mapping of a linear epitope on EDIII of Japanese encephalitis virus using a novel neutralizing monoclonal antibody

Wen Lei Deng, Chi Yu Guan, Ke Liu, Xiao Min Zhang, Xiuli Feng, Bin Zhou, Xiao Dong Su, Pu Yan Chen

Research output: Contribution to journalArticlepeer-review

6 Scopus citations


The domain III (EDIII) of the envelope protein of Japanese encephalitis virus (JEV) is proposed to play an essential role in JEV replication and infection; it is involved in binding to host receptors and contains specific epitopes that elicit neutralizing antibodies. However, most previous studies have not provided detailed molecular information about the functional epitopes on JEV EDIII protein. In this study, we described a monoclonal antibody (mAb 2B4) we produced and characterized by IFA, PRNT, ELISA and Western blot analyses. The results showed that mAb 2B4 was specific to JEV EDIII protein and possessed high neutralization activity against JEV in vitro. Furthermore, we found that the motif, 394HHWH397, was the minimal unit of the linear epitope recognized by mAb 2B4 through screening a phage-displayed random 12-mer peptide library. Using sequence alignment analysis it was found that this motif was highly conserved among JEV strains and was present in West Nile Virus (WNV). Indeed, ELISA data showed that this epitope could be recognized by both JEV-positive swine serum and WNV-positive swine serum. Notably, this linear epitope was highly hydrophilic and was located within the terminal end of a β-pleated sheet of EDIII. An analysis of the spatial conformation supported the possibility of inducing specific antibodies to this epitope. Taken together, we identified 394HHWH397 as an EDIII-specific linear epitope recognized by mAb 2B4, which would be beneficial for studying the pathogenic mechanism of JEV; and mAb 2B4 was also a potential diagnostic and therapeutic reagent.

Original languageEnglish (US)
Pages (from-to)133-139
Number of pages7
JournalVirus Research
Issue number1
StatePublished - Jan 22 2014
Externally publishedYes


  • EDIII protein
  • Epitope
  • Japanese encephalitis virus
  • Monoclonal antibody
  • Spatial conformation

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases
  • Cancer Research


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