Monoclonal antibody to N protein of porcine epidemic diarrhea virus

Monoclon Antib Immunodiagn Immunother. 2015 Feb;34(1):51-4. doi: 10.1089/mab.2014.0062.

Abstract

The N gene of porcine epidemic diarrhea virus (PEDV) was amplified by RT-PCR using specific primers, and inserted into the expression vector pCold-I to construct a recombinant plasmid pCold-I-N. The recombinant plasmid was expressed in Escherichia coli BL21 (DE3) under IPTG induction. Then, female BALB/c mice were immunized with the purified recombinant N protein and one strain of hybridoma cells named 2B8 secreting anti-N protein monoclonal antibodies (MAb) was obtained by hybridoma technique. The MAb was specifically reacted with PEDV and identified by Western blot and indirect immunofluorescence assays. This work indicated that the MAb would be a valuable tool as a specific diagnostic reagent for PEDV epidemiological surveys and diagnosis in the future.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / isolation & purification
  • Antibodies, Viral / biosynthesis*
  • Antibodies, Viral / isolation & purification
  • Antibody Specificity
  • Blotting, Western
  • Cell Line
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / immunology
  • Coronavirus Infections / veterinary*
  • Coronavirus Infections / virology
  • Coronavirus Nucleocapsid Proteins
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Founder Effect
  • Gene Expression
  • Hybridomas / immunology
  • Mice
  • Mice, Inbred BALB C
  • Nucleocapsid Proteins / chemistry
  • Nucleocapsid Proteins / genetics
  • Nucleocapsid Proteins / immunology*
  • Porcine epidemic diarrhea virus / immunology
  • Porcine epidemic diarrhea virus / isolation & purification*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / immunology
  • Swine Diseases / virology

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Coronavirus Nucleocapsid Proteins
  • Nucleocapsid Proteins
  • Recombinant Proteins