Production and characterization of virus-like particles and the P domain protein of GII.4 norovirus

J Virol Methods. 2012 Jan;179(1):1-7. doi: 10.1016/j.jviromet.2011.05.009. Epub 2011 May 12.

Abstract

Noroviruses are an important cause of epidemic acute gastroenteritis in humans. In this study the production and characterization of GII.4 norovirus virus-like particles (VLPs) in insect cells is reported. Furthermore, the expression of corresponding norovirus polyhistidine-tagged P domain protein in Escherichia coli is described. The protruding P domain of the norovirus capsid is known to contain determinants for antibody and receptor binding. Therefore, P domain proteins were studied as an alternative diagnostic tool for evaluating norovirus infection. Analyses by dynamic light scattering and cryo-electron microscopy revealed the presence of intact VLPs with an average diameter of about 40 nm. Immunostaining and ELISA assays using norovirus-specific human sera revealed that VLPs and the P domain are recognized by norovirus-specific antibodies and by their putative receptor. The VLPs and P domain protein are potentially useful in the development of diagnostic and vaccination tools for noroviruses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / blood
  • Caliciviridae Infections / diagnosis
  • Caliciviridae Infections / prevention & control
  • Cell Line
  • Escherichia coli / genetics
  • Gene Expression
  • Humans
  • Immunoassay
  • Norovirus / genetics*
  • Norovirus / immunology*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Spodoptera
  • Viral Proteins / genetics*
  • Viral Proteins / immunology*
  • Viral Vaccines / immunology
  • Virosomes / genetics
  • Virosomes / immunology*
  • Virosomes / isolation & purification*
  • Virosomes / metabolism

Substances

  • Antibodies, Viral
  • Recombinant Proteins
  • Viral Proteins
  • Viral Vaccines
  • Virosomes