Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein

Vet Microbiol. 2008 Sep 18;131(1-2):73-81. doi: 10.1016/j.vetmic.2008.02.022. Epub 2008 Mar 4.

Abstract

S1D (residues 636-789) is a neutralizing epitope region on the spike protein (S) of porcine epidemic diarrhea virus (PEDV). To accurately identify epitopes on S1D, the S1-phage library containing the gene encoding the S1D region of PEDV S protein was micropanned by six specific monoclonal antibodies (McAbs) against the S1D region. These micropanned epitope regions (MER) were focused on 696-779 amino acids of the S protein. To further map epitopes of the MER, seven overlapping mini-fragments covering MER nucleotides were separately synthesized and expressed in Escherichia coli BL21 with a GST tag. These mini-GST fusion proteins were scanned by ELISA and Western blotting with the six McAbs, and the result showed that S1D5 (residues 744-759) and S1D6 (residues 756-771) are two linear epitopes of the PEDV S protein. The antisera of the epitopes S1D5 and S1D6 could react with the native S protein of PEDV. Furthermore, Pepscan of the two linear epitopes demonstrated that SS2 ((748)YSNIGVCK(755)) and SS6 ((764)LQDGQVKI(771)) are two core epitopes on S1D5 and S1D6, respectively, located on the S protein of PEDV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Base Sequence
  • Blotting, Western / veterinary
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Epitope Mapping
  • Epitopes, B-Lymphocyte / immunology*
  • Gastroenteritis, Transmissible, of Swine / immunology*
  • Gastroenteritis, Transmissible, of Swine / virology*
  • Molecular Sequence Data
  • Neutralization Tests / veterinary
  • Porcine epidemic diarrhea virus / genetics
  • Porcine epidemic diarrhea virus / immunology*
  • Swine
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / immunology*

Substances

  • Antibodies, Monoclonal
  • Epitopes, B-Lymphocyte
  • Viral Envelope Proteins