Identification of Properties that Affect in vitro Virus-Like Particle Assembly

Natalie Connors (2011). Identification of Properties that Affect in vitro Virus-Like Particle Assembly PhD Thesis, Aust Institute for Bioengineering & Nanotechnology, The University of Queensland.

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
s33638700_phd_final_abstract.pdf PhD Abstract Final application/pdf 9.00KB 3
s33638700_phd_final_thesis.pdf PhD Thesis Final application/pdf 8.18MB 18
Author Natalie Connors
Thesis Title Identification of Properties that Affect in vitro Virus-Like Particle Assembly
School, Centre or Institute Aust Institute for Bioengineering & Nanotechnology
Institution The University of Queensland
Publication date 2011-11
Thesis type PhD Thesis
Total pages 264
Total colour pages 41
Total black and white pages 223
Language eng
Subjects 060112 Structural Biology (incl. Macromolecular Modelling)
060102 Bioinformatics
Abstract/Summary Virus-like particles (VLPs) are biological nanoparticles formed from the self-assembly of viral subunits into a shell or capsid, which lacks the infectious DNA or RNA genome of the virus, and are presently one of the major focal points for therapeutic delivery systems research. Predominantly, the focus of VLPs is largely based on vaccine technology, with several highly efficacious VLP-based vaccines currently on the market to prevent human disease including Hepatitis B and Cervical Cancer. The self-assembling property of VLPs has lead to the development of a cell-free bioprocessing route utilising recombinant protein technology and chemical self-assembly. However, current understanding of this bioprocess is poor, with inadequate control over ordered assembly, and often resulting in VLPs with poor architectural consistency and low quaternary structure stability. Here we have taken a process-inspired biomolecular engineering approach to this processing problem. There are three key parts to this VLP bioprocessing challenge that are addressed within this project: (i) quaternary structure stability, (ii) architectural consistency and homogeneity, and (iii) self-assembly. We were able to use computational analyses to direct mutagenesis studies for a better understanding of VLP quaternary structure, and alter the proteolytic profile of the major coat protein to address sample heterogeneity. This included mutagenesis of known and predicted proteolytic cleavage sites, and the introduction of intercapsomeric disulfide bonding. We also present the first known second virial coefficient computational model for the viral assembly process. This study has enabled a greater understanding of the structure-function relationships for the VLP proteins and highlighted both the rigidity and flexibility the protein has to mutagenesis. This information is key for future modifications to the VLP for vaccine development potential. In addition, fundamental understanding of the mechanisms controlling the self-assembly process has been probed here with the second virial coefficient model. Exploration of the intermolecular interactions between assembling subunits will provide a knowledge base, whereby we can predict the optimal conditions for VLP assembly. Ultimately there is the potential to create a generic VLP, applicable to a range of biological problems or diseases, generating a platform technology that can be manipulated or adapted to suit the required function of the VLPs. The key to enable this platform technology is to understand the VLP’s response to modification, and understand the self-assembly. This project contributes to this understanding.
Keyword Virus-like Particles
Murine Polyomavirus
Second virial coefficient
self assembly
disulfide engineering
protein conservation
Additional Notes 20, 23, 33-35, 37-39, 64, 82, 88, 89, 92, 93, 97, 98, 100, 102, 106, 109, 130, 132, 133, 136, 144, 145, 149, 170, 175, 186, 191, 255-264.

Citation counts: Google Scholar Search Google Scholar
Created: Fri, 04 May 2012, 09:10:04 EST by Ms Natalie Connors on behalf of Library - Information Access Service