Scanning Gel Chromatograpy
Gel permeation chromatography is widely used both as a preparative separation technique and as an analytical tool for the estimation of molecular size and the study of interactions between macromolecules. In this latter category it is unsurpassed among physical techniques in sensitivity, precision, and flexibility. The migration rates and detailed shapes of zones containing interacting solutes permit the determination of reaction stoichiometry and equilibrium constants over a very broad range of conditions. In addition to these nonequilibrium transport methods, the technique of equilibrium gel permeation provides a powerful means to study thermodynamic properties of multicomponent interacting solute systems, such as macromolecule-ligand binding (see Ackers, 1975, for a general review). A new and particularly promising development is the technique of active enzyme chromatography (Jones, 1975; Jones et al., 1976), in which a small band of enzyme is passed down a column previously saturated with substrate and the product of the enzymatic reaction is monitored by successive optical scanning of the column during transport. By determining the “rate of motion” of the chromophoric product being formed, it is possible to infer the molecular size of the active species.
KeywordsPartition Coefficient Methyl Orange Axial Dispersion Solute Profile Molecular Radius
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