Grant: Research Tools and Instruments - Category 1 (<$150,000)

Our entire research program is concerned with the development of novel approaches for kinetic studies of non-covalent biomolecular interactions. Accordingly, proposed instrumentation for label-free solution-based kinetic analysis of such interactions is pivotal. The kinetics of biomolecular interactions control the dynamics of cellular processes, such as DNA replication and gene expression. They define the efficiency of analytical techniques, such as immunoassays and hybridization assays. They also define pharmacokinetics of drugs whose action is based on drugs' ability to form complexes with their therapeutic targets. Conventional methods for kinetic studies of biomolecular interactions require either the immobilization of one of the binders on the sensor or its labeling with a fluorophore. Not only are the immobilization and labeling difficult to achieve, but they can also drastically affect the kinetics. We pioneered Kinetic Capillary Electrophoresis (KCE) with Mass Spectrometry (MS) detection, a solution-based label-free approach to study biomolecular interactions. The approach was proved in-principle by using a tedious ""off-line"" mode. Backman-Coulter has recently invented and started marketing a sheathless interface between Capillary Elctrophoresis (CE) and MS which can facilitate simple ""on-line"" kinetic studies. Here we propose to assemble an instrument for on-line KCE-MS and use this instrument in multiple projects concerned with kinetics of biomolecular interactions. Several MS instruments are available to us; therefore, we only request support for purchasing a CE instrument with a sheathless interface. The research program is supported by NSERC Strategic, NSERC Discovery, NSERC Discovery Accelerator, and NSERC I2I grants in total exceeding $500,000 per year. Five graduate students and four postdoctoral fellows are currently working on kinetic aspects of biomolecular interactions. Training of these researchers critically depends on the availability of the proposed instrument. If the laboratory does not get the instrument in 2014 then our research in many directions will be impeded and in others completely stalled. It will also cause delays in completion of degree requirements by graduate students.