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Protein Sci. 2016 Aug;25(8):1507-16. doi: 10.1002/pro.2953. Epub 2016 Jun 13.

Design strategies to address the effect of hydrophobic epitope on stability and in vitro assembly of modular virus-like particle.

Author information

1
The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Centre for Biomolecular Engineering, St Lucia, Queensland 4072, Australia.
2
The University of Queensland, UQ Protein Expression Facility, University of Queensland, St Lucia, Queensland 4072, Australia.

Abstract

Virus-like particles (VLPs) and capsomere subunits have shown promising potential as safe and effective vaccine candidates. They can serve as platforms for the display of foreign epitopes on their surfaces in a modular architecture. Depending on the physicochemical properties of the antigenic modules, modularization may affect the expression, solubility and stability of capsomeres, and VLP assembly. In this study, three module designs of a rotavirus hydrophobic peptide (RV10) were synthesized using synthetic biology. Among the three synthetic modules, modularization of the murine polyomavirus VP1 with a single copy of RV10 flanked by long linkers and charged residues resulted in the expression of stable modular capsomeres. Further employing the approach of module titration of RV10 modules on each capsomere via Escherichia coli co-expression of unmodified VP1 and modular VP1-RV10 successfully translated purified modular capomeres into modular VLPs when assembled in vitro. Our results demonstrate that tailoring the physicochemical properties of modules to enhance modular capsomeres stability is achievable through synthetic biology designs. Combined with module titration strategy to avoid steric hindrance to intercapsomere interactions, this allows bioprocessing of bacterially produced in vitro assembled modular VLPs.

KEYWORDS:

Escherichia coli; co-expression; linkers; module titration; rotavirus; synthetic biology

PMID:
27222486
PMCID:
PMC4972206
DOI:
10.1002/pro.2953
[Indexed for MEDLINE]
Free PMC Article

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