Dr Robert J Edwards

Dr Robert Edwards is a Research Lecturer in the Department of Experimental Medicine and Toxicology. His current research interests include investigation of protein biomarkers and the development of a universally-applicable approach for the generation of protein-specific antibodies.

Protein biomarkers: Modern techniques of protein analysis and identification are being used to discover new protein biomarkers of disease and toxicity. Such biomarkers may be used to improve diagnosis, provide methods for earlier detection of disease, and to give an insight into the disease process. A variety of projects are underway that are based on the use of two main proteomic techniques. These are SELDI-TOF (surface-enhanced laser desorption ionisation time of flight) mass spectrometry and label-free quantitative proteomics. SELDI-TOF mass spectrometry is rapid in its ability to produce protein profiles from complex mixtures and provide a means to compare relative levels of protein expression; the identity of proteins of interest is then established after purification. This has been done successfully for several proteins. Label-free quantitative proteomics requires pre-fractionation of samples prior to analysis by nanoflow liquid chromatography coupled to electrospray ionisation mass spectrometry. This approach yields protein identification data directly and has been successfully applied to an analysis of estrogen-responsive proteins in breast cancer cells and is currently being applied to the analysis of cultured neuronal cells, skin, lung and plasma samples from patients with cardiovascular diseases.

Development of a universally-applicable approach for the generation of protein-specific antibodies: A long-term interest in the use of anti-peptide antibodies has resulted in the development of a novel approach to the efficient production of highly specific antibodies against proteins. The approach, which is based on targeting a minimal epitope at the C-terminus of proteins to produce a CTAb (C-terminal antibody) has been shown to have a number of advantages over the use of monoclonal antibodies or polyclonal antibodies raised against purified proteins. These include the high degree of specificity of the antibodies, the ease of production and the ability to direct antibodies to pre-determined regions of the target protein. As the antibodies bind equally well to native and denatured proteins they are suitable for use in a wide range of biochemical techniques, including western blotting, immunoprecipitation and immunohistochemistry. CTAbs against P450 enzymes from human and other species have been raised, as well as against other proteins, such as theta-glutathione transferase, histone H2B, chemokine receptor CCR5, and human α-tropomyosin. This approach has recently been extended successfully to target a variety of proteins expressed by Streptococcus pyogenes and thus demonstrates its broad utility. Raising antibodies in this way is fast and efficient and thus meets the increasing need for suitable antibodies necessary in follow up studies to support the proteomic identification of proteins of interest.