Kinetic capillary electrophoresis - an analytical Swiss army knife
Chemistry, York University
(April 14, 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 (kon, koff, Kd, Delta H, and Delta S) 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 [1-10]. I 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.
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