Development of diagnostic platform for biomolecular interactions and biocatalysis using capillary electrophoresis and mass spectrometry

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There are two main goals of our application to the Collaborative Research and Development Grant. The first goal is to develop a new platform for screening enzymes, substrates and inhibitors based on capillary electrophoresis (CE) coupled to electrospray ionization mass spectrometry (MS). CE-MS requires only nanoliters of reagent solutions and uses the separation capillary as a microreactor, allowing multiple substrates to be assayed simultaneously. The method can be used to rapidly profile the substrate specificity of any enzyme and to measure steady-state kinetics in an automated fashion. The second goal is to develop a new diagnostic platform to study protein dynamics and affinity interactions based on kinetic CE-MS. Kinetic CE-MS establishes a new paradigm that separation methods together with MS are used as comprehensive kinetic tools with mass and structure elucidation in proteomics and genomics. Kinetic CE-MS serves as a valuable platform for studying protein structure and function, including the in situ catalytic activity of conformers and their susceptibility to inhibitor binding.We aim to combine a CE instrument made by LUMEX Instruments Canada in Canada with a mass spectrometer in one diagnostic platform. The successful completion of this collaborative project between uOttawa and LUMEX is expected to generate new technologies based on the capillary electrophoresis platforms that will find wide application in diagnostics, drug discovery, enzyme development, antibody screening and will impact both Canadian industry and the well-being of Canadians.