Purification of a baculovirus-expressed hepatitis E virus structural protein and utility in an enzyme-linked immunosorbent assay

J Clin Microbiol. 1995 Dec;33(12):3308-11. doi: 10.1128/jcm.33.12.3308-3311.1995.

Abstract

We report on the purification of the full-length structural protein encoded by open reading frame 2 (ORF-2) of hepatitis E virus. The ORF-2 protein, expressed in Sf9 cells by using a recombinant baculovirus vector system, was successfully purified to homogeneity. Gel electrophoresis of the purified ORF-2 protein showed a single polypeptide of 75 kDa by Coomassie blue staining and by Western blot (immunoblot) analysis. We demonstrated that the partially purified ORF-2 protein could be used successfully in a sensitive and specific enzyme-linked immunosorbent assay for the detection of antibodies to hepatitis E virus.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Gene Expression
  • Genetic Vectors
  • Hepatitis Antibodies / blood
  • Hepatitis E / diagnosis
  • Hepatitis E / immunology
  • Hepatitis E virus / genetics*
  • Hepatitis E virus / immunology*
  • Humans
  • Immunoglobulin M / blood
  • Nucleopolyhedroviruses / genetics
  • Open Reading Frames
  • Protein Folding
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Spodoptera
  • Viral Structural Proteins / genetics*
  • Viral Structural Proteins / immunology*
  • Viral Structural Proteins / isolation & purification

Substances

  • Hepatitis Antibodies
  • Immunoglobulin M
  • Recombinant Proteins
  • Viral Structural Proteins