Kinetic Separation: A Conceptual Platform for Development of Homogeneous Kinetic Affinity Methods

Synopsis

Non-covalent binding of biological molecules controls cellular regulation, it is also pivotal to the action of modern drugs and molecular diagnostics of diseases. Therefore, understanding molecular mechanisms of biological processes as well as the development of drugs and design of diagnostic systems require affinity methods suitable for three types of applications: (i) kinetic studies of affinity interactions (ii) selection of affinity ligands from combinatorial libraries and iii) the use of affinity ligands as diagnostic probes in molecular diagnostics of diseases.

Our answer to this challenge is Kinetic Separation - a conceptual platform for the development of homogeneous kinetic affinity methods suitable for all three applications. One-dimensional (column) separation of any nature, e.g. electrophoresis, gel-filtration, thermophoresis, sedimentation, that can segregate affinity complexes from the unbound interactants, can be used as the basis for kinetic separation. A variety of different detection methods can be employed in kinetic separation including fluorescence and mass-spectrometry.

In this lecture, Prof Krylov will explain the physical properties underlying kinetic separation and discuss two practical implementations of kinetic separation for (i) selection, kinetic characterisation, and analytical use of DNA aptamers by means of Kinetic Capillary Electrophoresis and (ii) studying kinetics of reversible protein-drug binding by means of Kinetic Size-Exclusion Chromatography.

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Speaker

Prof Sergey Krylov

Centre for Research on Biomolecular Interactions, York University in Toronto, Canada