New method for immobilising diverse proteins onto cubic micro-protein polyhedrin crystals

Protein Expr Purif. 2020 Mar:167:105531. doi: 10.1016/j.pep.2019.105531. Epub 2019 Nov 14.

Abstract

Cypovirus is an insect virus that is encapsulated in stable cubic protein crystals composed of polyhedrin protein produced in virus-infected cells. Molecular technology developed over the last decade is now able to immobilise proteins of interest on polyhedrin crystals. Modified polyhedrin crystals can be used in cell cultures for implantation in animals and vaccines, among other applications. However, this technique does not work for some proteins. Here, we developed and tested an alternative approach for immobilising foreign proteins in polyhedrin crystals using a linker method; diverse proteins, such as fluorescent proteins, enzymes, antibodies, and streptavidin were successfully contained. The immobilised antibodies retained their binding activity on filter paper, implying their potential for new immunochromatography applications. Moreover, this immobilisation method allows enzymes to be collected from one reaction reagent and transferred to another reagent. These results demonstrate the potential of this immobilisation method and the likelihood of expanding the applications of polyhedrin crystals using this approach.

Keywords: Cubic protein crystals; Polyhedrin; Protein immobilisation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Immobilized Proteins / chemistry*
  • Occlusion Body Matrix Proteins / chemistry*
  • Protein Engineering / methods
  • Reoviridae / chemistry
  • Viral Proteins / chemistry
  • Viral Structural Proteins / chemistry

Substances

  • Immobilized Proteins
  • Occlusion Body Matrix Proteins
  • Viral Proteins
  • Viral Structural Proteins