Engineering of a specific binding site for protein labelling with luminescent lanthanide coated nanoparticles: A study of protein labelling and nanoparticles

Project completed 2013.

Supervisors:
Professor Zoe Pikramenou, School of Chemistry
Professor Mike Hannon, School of Chemistry
Dr Josh Rappoport, School of Biosciences
Professor John Heath, School of Biosciences
Professor Ela Claridge, School of Computer Science

Gold and platinum nanoparticles provide a unique scaffold for the co-attachment of fluorescent agents and targeting vectors for delivery into cells. In addition lanthanide coated nanoparticles have properties that make them attractive probes for use in single cell and organism imaging studies. Recent results, based on a range of imaging techniques, have shown that lanthanide coated nanoparticles can be delivered into cells. However, to fully exploit the potential of nanoparticles in cells, tissues and organisms, it is necessary to understand and develop chemical methods for their conjugation to biologically active probes and how this affects delivery into cells.

This project aims to develop a new molecular probe for generic protein labelling. Progression will be dependent on the following practical elements:

- Development of a suitable peptide binding site to preferentially bind to nanoparticles in vivo.

- Establishment of cell locations accessible to nanoparticles using live cell fluorescence microscopy and transmission electron microscopy.

- Development of protocol to transfect cells for the expression of protein-peptide binding site complexes.

- Imaging of protein labelling by nanoparticles carrying different probes in vivo.

The research will include analysis of the success of nano-probes by using a range of light and electron microscope methods on cell lines treated with the compounds as well as by using analytical techniques such as STEM and ICP-MS. Once this is established the project will extend to the labelling and/or delivery of other molecules into cells, including anti-cancer drugs, antibodies and biomolecular compounds.

Link to ethesis: http://etheses.bham.ac.uk/5576/