title: Synthetic biology design to display an 18 kDa rotavirus large antigen on a modular virus-like particle.
abstract: Virus-like particles are an established class of commercial vaccine possessing excellent function and proven stability. Exciting developments made possible by modern tools of synthetic biology has stimulated emergence of modular VLPs, whereby parts of one pathogen are by design integrated into a less harmful VLP which has preferential physical and manufacturing character. This strategy allows the immunologically protective parts of a pathogen to be displayed on the most-suitable VLP. However, the field of modular VLP design is immature, and robust design principles are yet to emerge, particularly for larger antigenic structures. Here we use a combination of molecular dynamic simulation and experiment to reveal two key design principles for VLPs. First, the linkers connecting the integrated antigenic module with the VLP-forming protein must be well designed to ensure structural separation and independence. Second, the number of antigenic domains on the VLP surface must be sufficiently below the maximum such that a "steric barrier" to VLP formation cannot exist. This second principle leads to designs whereby co-expression of modular protein with unmodified VLP-forming protein can titrate down the amount of antigen on the surface of the VLP, to the point where assembly can proceed. In this work we elucidate these principles by displaying the 18.1 kDa VP8* domain from rotavirus on the murine polyomavirus VLP, and show functional presentation of the antigenic structure.
authors.affiliation: The University of Queensland, Protein Expression Facility, St Lucia, QLD 4072, Australia.
authors.forename: Linda H L
authors.initials: LH
authors.lastname: Lua
authors.affiliation: The University of Queensland, Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Fan
authors.affiliation: The University of Queensland, Protein Expression Facility, St Lucia, QLD 4072, Australia.
authors.forename: Cindy
authors.initials: C
authors.lastname: Chang
authors.affiliation: The University of Queensland, Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia.
authors.forename: Natalie K
authors.initials: NK
authors.lastname: Connors
authors.affiliation: The University of Queensland, Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia. Electronic address: a.middelberg@uq.edu.au.
authors.forename: Anton P J
authors.initials: AP
authors.lastname: Middelberg
mesh_terms: D000956:Antigens, Viral / Q000737:chemistry* / Q000235:genetics; D005822:Genetic Vectors; D056004:Molecular Dynamics Simulation; D011120:Polyomavirus / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D012401:Rotavirus / Q000235:genetics*; D022243:Rotavirus Vaccines / Q000737:chemistry* / Q000235:genetics; D058425:Vaccines, Virus-Like Particle / Q000737:chemistry* / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D022243:Rotavirus Vaccines; D058425:Vaccines, Virus-Like Particle
keywords: Computational; Polyomavirus; Rotavirus; Vaccine; Virus-like particle