Affinity Chromatography, Adsorption Chromatography, Gel Filtration Chromatography, Spacer Arms

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Affinity Chromatography
In this method, enzymes are purified according to their specificity for a particular substrate or cofactor. One component of the mixture, containing the enzyme, binds covalently to the solid support of the column, and the other components percolate down through the column. The basic requirements for affinity chromatography are the same as in adsorption or gel filtration chromatography but the packing gel must have some component which can bind with one component of the mixture.

Many commercial gels available for affinity chromatography contain functional groups attached to the 'spacer arms' of the gel. The spacer arm is a chemical linkage between the functional group and the gel or matrix proper, so that the binding between functional group and the enzyme is kept away from the gel. Thus the steric hindrance will be unlikely to prevent binding of the specific enzyme to the column.

An Arrangement for Affinity Chromatography, Showing the Role of Functional Group having Affinity with the Enzyme

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