Display of Viral Surface Glycoprotein on Virus like particles (VPLS) for Structure and Immune Epitope Discovery

Summary

Unanchored recombinant soluble monomers are often the proteins used in many structure analyses and immunologic testing. But in reality viral surface glycoproteins are orientated, tethered and displayed on the viral surface as protein complexes, quite often as homo-trimers. The aim is the production of Virus Like Particles for the display of expressed viral surface proteins in near to native embedded state, complex and configuration, therefore highly suitable for quality downstream structure and function analysis and antibody binding studies of value for vaccine design and discovery.

Supervisor(s)

Dr Garry Lynch

Research Location

Camperdown - Central Clinical School

Program Type

Masters/PHD

Synopsis

One of the most significant roadblocks in the study of viral and bacterial surface proteins is in many cases a lack of defined high-resolution structures. Using bioinformatics and molecular modeling we have been able to reveal the surface protein structures for a number of human and animal viruses (eg Pandemic Influenza, Ebola, HIV, Hendra, Rabies). To complement those studies, Virus-like particles (VLP) are to be prepared, and used to express viral surface antigens in a manner that is fully non-infectious and safe for direct structural studies. The challenge in this study will be to prepare VLP, which are then deployed to display an expressed glycoprotein(s), such as the gp proteins that adorn the surfaces of the hemorrhagic Ebola and Marburg viruses. This would firstly be of benefit for the production of viral envelope proteins, as they naturally exist in complex configurations on the viral surface. This VLP project therefore permits structure function analysis in near native configurations, and to study their structures and interactions biochemically and by cryo-electron microscopy. This would also facilitate antibody screening and capture, and enable comparative studies of strain-specific and broad-strain antibodies. Importantly glycoprotein thus decorated on the surface of VLPs would provide a significant means for the study of anti-viral antibody binding interactions in close-to native states and configurations, directly and in fusion and cell entry assays. Each of the above would particularly assist our efforts in developing broadly acting vaccine constructs for pathogenic viruses. For biomed students with art based skills and interests there is scope for some BioVisualization of the study viruses and the tissues they infect etc. This VLP project is to be a new but highly valuable direction for our studies and would suit a student who likes a challenge, and has initiative to be creative and make it happen.

Collaborators: Dr William Bret Church (Pharmacy), A/Prof John Sullivan (Medicine), Prof Hans Zoellner (Dentistry), A/Prof Peter Williamson (SOLES, Science).

Additional Information

Techniques can involve: Proteomics, cell and protein fractionation, electrophoresis, immunoblotting, immunoprecipitation, multiplex-analysis. Bioinformatics, computational modelling, analysis and design of protein structures. Computational and wet laboratory studies of antibodies and antibody-binding, recombinant proteins, peptide studies and analyses; mass spectrometry, x-ray crystallisation, cryoelectron microscopy, surface plasmon resonance, molecular biology, protein expression.
Relevant References:
1. Lynch, G et al Seasoned adaptive antibody immunity for highly pathogenic pandemic influenza in humans. Immunol Cell Biol 90: 149-158. (2012).
2. Lynch, G.W., Selleck, P. & Sullivan, J.S. Acquired heterosubtypic antibodies in human
immunity for avian H5N1 influenza. J Mol Genet Med 3, 205-209 (2009).
3. Lynch, G.W. et al. Cross-Reactive anti-Avian H5N1 Influenza Neutralizing Antibodies in a
Normal ‘Exposure-Naïve' Australian Blood Donor Population. The Open Immunol J 1 (2008).
4. Lynch G.W et al. Imaging of High and Low Resolution Ebola Envelope GP Structures Composited with in silico Models of Difficult-to-Resolved Sections. J. Molecular & Genetic Medicine 9 (4): 186-92, 2015

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Keywords

virus, infection, Infectious Disease, Ebola, Marburg, protein, structure, gp, glycoprotein, Outbreak, Immunology, Adaptive Immunity, Vaccine, Protection

Opportunity ID

The opportunity ID for this research opportunity is: 2592

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