Project No. 2140

Primary Supervisor

Dr Iris Nandhakumar – University of Southampton

Co-Supervisor(s)

Dr Jurgita Zekonyte – University of Portsmouth

Summary

This project focuses on the development of a ground breaking and novel bioanalytical technique that will enable the functional imaging of cells and therefore fill a major technology gap in the biologist’s toolkit.

Structure-function relationships are central to our greater understanding of the biological world. Currently, these relationships are frequently approached using techniques such as X-ray crystallography and NMR. However cell surface proteins and receptors are embedded within the lipid bilayer of the plasma membrane and so are not amenable to analysis with these current modalities. Furthermore, crystallography and NMR do not provide information on whole cells in real time and so are unable to address complex binding interactions at the cell surface. There is therefore, a real need to develop new technologies which are able to address these key questions. In this application we aim to combine the high specificity of antibodies and the sensitivity and flexibility of atomic force microscopy in order to develop novel technology in order to address these questions.

The Atomic Force Microscope (AFM) has become an indispensable tool for studying interfacial phenomena and has greatly impacted upon many areas of science and technology due to its unique ability to image and characterize structures in liquid, ambient and vacuum environments. The functionality of AFM can be dramatically extended in chemical force microscopy (CFM) that adds the capability of chemical discrimination and involves the modification of AFM probes with specific functional groups (chemical or biological) to detect chemical contrast simultaneously with topography during AFM imaging.

Monoclonal antibodies (mAb), with their ability to bind individual target molecules (proteins, enzymes, carbohydrates, lipids etc.) with high specificity and affinity are ideally suited for this purpose. In fact, they have transformed many aspects of scientific endeavour, facilitating molecular isolation, purification and identification as well being exploited for diagnostics and front-line therapy of disease. Therefore, the combination of these two technologies is clearly appealing and will allow the interrogation of cell surface interactions at a level of specificity, real-time dynamism and resolution previously impossible.