ABSTRACT
Enzyme purification involves the separation of a single protein from a mixture which may contain over a thousand components. Prior to the 1950s, most protein purification procedures were based upon changing ionic strength, dielectric constant, or other solution properties. Development of more specialized techniques such as isoelectric focusing, preparative electrophoresis, and affinity chromatography in the last 20 years has proven valuable for separation of many enzymes. Preparation of high-performance cation exchange chromatography supports has lagged behind the development of the anion exchange materials. As indicated by its name, the process of ion-exchange chromatography involves interaction of the solute with the surface of the support. Diethylaminoethyl (DEAE)-cellulose purified hexokinase loses the ability to rebind mitochondria, but hexokinase purified by high-performance ion exchange chromatography (HPIEC) chromatography retains this characteristic. Also, the HPIEC column can separate the bindable form of the enzyme from the non-bindable form which results from DEAE-cellulose chromatography.
