Novel expression of coat proteins from thermophilic bacteriophage ΦIN93 and evaluation for assembly into virus-like particles

Protein Expr Purif. 2021 Nov:187:105932. doi: 10.1016/j.pep.2021.105932. Epub 2021 Jun 29.

Abstract

Virus-like particles (VLPs) have the potential to be used as display platforms to develop vaccines against infectious and non-infectious agents. However, most VLPs used as vaccine display platforms are derived from viruses that infect humans; unfortunately, most humans already have pre-existing antibodies against these platforms and thus, the immunogenicity of these vaccines may be compromised. VLP platforms derived from viruses that infect bacteria (bacteriophages), especially bacteriophages that infect bacteria, which do not colonize humans are less likely to have pre-existing antibodies against the platforms in the human population. In this study, we assessed whether two putative coat proteins (ORF13 and ORF14) derived from a thermophilic bacteriophage (ΦIN93) can be expressed and purified from a mesophilic bacterium such as E. coli. We also assessed whether expressed coat proteins can assemble to form VLPs. Truncated versions of ORF13 and ORF14 were successfully co-expressed in bacteria; the co-expressed truncated proteins formed oval structures that look like VLPs, but their sizes were less than those of an authentic ΦIN93 virus.

Keywords: Assembly; Coat proteins; Expression; Thermophilic bacteriophage; Virus-like particles.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Neutralizing / chemistry
  • Antibodies, Neutralizing / genetics
  • Antibodies, Neutralizing / metabolism
  • Bacterial Infections / metabolism
  • Bacteriophages / metabolism*
  • Capsid Proteins / chemistry
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism*
  • Escherichia coli
  • Gene Expression Regulation
  • Humans
  • Protein Binding
  • Vaccines, Virus-Like Particle / chemistry
  • Vaccines, Virus-Like Particle / metabolism*
  • Viruses / genetics
  • Viruses / metabolism*

Substances

  • Antibodies, Neutralizing
  • Capsid Proteins
  • Vaccines, Virus-Like Particle