New tools to provide mathematical modellers of molecular processes with rate constants of biomolecular interaction
Sergey Krylov (Chemistry, York University)
(May 12, 2009 2:30 PM - 3:30 PM)
Kinetic capillary electrophoresis (KCE) is defined as capillary electrophoresis of species that interact during electrophoresis. KCE can serve as a conceptual platform for development of homogeneous kinetic affinity methods for affinity measurements and affinity purification. Affinity measurements include two groups of applications: (i) measurements of kinetic and thermodynamic parameters (k_on, k_off, K_d, deltaH, and deltaS) of affinity interactions and (ii) quantitative measurements of concentrations using affinity probes (e.g. antibodies and aptamers). Affinity purification also includes two large groups of applications: (i) purification of known molecules and (ii) search of unknown molecules. A number of different KCE methods can be designed by varying initial and boundary conditions - the way interacting species enter and exit the capillary. The basic theory of KCE concept has been recently developed and a large number of practical applications have been proven. In this lecture, I will present the theoretical bases of KCE and explain how it can be used for: (i) measurements of rate and equilibrium constants of protein-DNA interaction, (ii) selection of smart aptamers, and (iii) using smart aptamers for protein detection in an ultra-wide range of concentrations.