Honours supervisors and projects

Dr Diane Webster

Plant-made recombinant proteins with therapeutic potential

Telephone: 9905 4343 or 0425 809 396
Email: diane.webster@sci.monash.edu.au

Traditional systems for the expression of recombinant proteins are hampered by high costs, problems with post-translational modifications, and human pathogen contamination risks. Plants represent an alternative production system that produces high quality recombinant protein, in an affordable, safe and scalable way. Plant-made human proteins have been shown to be fully functional and structurally comparable to native protein, with a similar pattern of glycosylation. Moreover, plant-based expression systems are free from contamination by pathogens.

Our research aims to express and characterize recombinant proteins in plants. Recombinant proteins are expressed by transient infiltration, stable transformation of plants and hairy root cultures and/or from viral vectors. Once characterized some proteins are then selectively modify to generate recombinant proteins with enhanced bioactivity, improved safety profiles, and increased yields. We have a particular interest in post-translational processing and its relationship to protein stability and function.

We are currently investigating a range of recombinant proteins with therapeutic potential, including vaccines for measles, malaria, hookworm and avian influenza (poultry vaccine), and the expression of proteins with potential therapeutic applications in diabetes, and heart disease. Projects may be co-supervised by Dr Amanda Walmsley and/or Prof John Hamill. Specific projects can be tailored to match the interest of individual students.

Some potential projects for 2009 include:

• Expression of hookworm proteins in plants (using transient and viral vector expression systems).
• Characterisation of the influenza HA protein produced in plants.
• Evaluation of the impact of glycosylation on plant-made measles virus HA protein stability – in planta, in vitro and in vivo.
• Investigation of the effect of post-translational modifications on receptor-ligand interactions and protein stability.
• Expression of scFv (antibody fragments) in plants (a potential therapeutic for heart disease).

Students will learn a range of techniques.  Depending on the project these may include; molecular biology (bacterial cloning, PCR, gel electrophoresis); plant transformation (transient agroinfiltration, stable transformation, hairy root cultures); protein analysis (ELISA, SDS-PAGE, Western blots, purification, glycosylation studies, in vitro functional assays); animal trials (animal handling, injection techniques, gavage and feeding); immunological analysis (ELISA, antibody isotyping, antibody affinity, IFNg ELISPOTs). Note; animal handling is optional for plant projects.