Generation of MERS-CoV Pseudotyped Viral Particles for the Evaluation of Neutralizing Antibodies in Mammalian Sera

Methods Mol Biol. 2020:2099:117-126. doi: 10.1007/978-1-0716-0211-9_10.

Abstract

Pseudotyped viral particle production has been used extensively and broadly for many viruses to evaluate levels of neutralizing antibodies, viral entry inhibitors and vaccine immunogenicity. This assay is extremely safe and useful alternative to live virus-based assay without the need for high containment facilities. In this chapter, we describe the generation of MERS-CoV pseudotyped viral particles (MERSpp) expressing full-length spike protein using second-generation lentiviral packaging system. This platform is optimized to generate high titer of MERSpp and to test sera from different mammalian species.

Keywords: MERS-CoV; Neutralization assay; Pseudotype viral particles; Pseudovirus; Transfection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood*
  • Coronavirus Infections / virology*
  • HEK293 Cells
  • Humans
  • Immunogenicity, Vaccine / immunology
  • Mammals
  • Middle East Respiratory Syndrome Coronavirus / immunology*
  • Middle East Respiratory Syndrome Coronavirus / isolation & purification
  • Neutralization Tests
  • Spike Glycoprotein, Coronavirus / genetics
  • Spike Glycoprotein, Coronavirus / immunology*
  • Spike Glycoprotein, Coronavirus / metabolism
  • Viral Vaccines / immunology*
  • Virion / genetics
  • Virion / immunology*
  • Virion / metabolism
  • Virus Internalization

Substances

  • Antibodies, Neutralizing
  • Spike Glycoprotein, Coronavirus
  • Viral Vaccines