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

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...

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Bibliographic Details
Published in:Protein expression and purification 2020-03, Vol.167, p.105531-105531, Article 105531
Main Authors: Yuasa, Haruna, Kotani, Eiji, Mori, Hajime, Takaki, Keiko
Format: Article
Language:English
Subjects:
Animals
Cubic protein crystals
Immobilized Proteins - chemistry
Occlusion Body Matrix Proteins - chemistry
Polyhedrin
Protein Engineering - methods
Protein immobilisation
Reoviridae - chemistry
Viral Proteins - chemistry
Viral Structural Proteins - chemistry
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Summary: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. •Polyhedrin protein cubic crystals can be a stage to immobilise foreign protein.•Several proteins were previously unable to be immobilised.•This new method allowed the crystals to immobilise diverse proteins.•This method will expand the application potential of polyhedrin protein crystals.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2019.105531