Development of capillary electrophoresis Methodologies to investigate the interactions of neutralizing agents with human rhinoviruses

Common cold viruses consist of a protein capsid which encloses an RNA genome. They thus exhibit a zeta potential giving rise to migration in an electric field. The migration behaviour is expected to be modified upon binding of antibodies, soluble virus receptors or small antivirally active compounds. This might occur either by changing the mass, masking the surface charge, modifying the conformation, or by a combination of all these effects. In the proposed project capillary electrophoresis techniques will be established to determine parameters such as binding constants and the stoichiometry of the complexes formed by the virus and the neutralizing agent. Appropriate techniques (such as affinity capillary zone electrophoresis or capillary isoelectric focusing) will be employed and the variables decisive for resolution will be investigated systematically. This work will result in an efficient and fast method allowing for separation of native virus from virus-ligand complexes. Amongst other results of these experiments we expect to establish a capillary electrophoretic method for the simple and rapid screening of various (low or high molecular weight) compounds for a potential neutralizing effect towards common cold viruses. In this context sensitive methods detecting conformational changes which are known to occur upon binding of certain antivirally active compounds will also be developed. Knowledge of the parameters influencing separation of these large macromolecular complexes will allow to extend the techniques to the investigation of the activity of antiviral agents against other viruses.