ABSTRACT
Ion-exchange chromatography is the most commonly used technique for isolating and characterizing different forms of cytochrome P-450. However, this technique is time consuming and the resolution and reproducibility of the chromatograms leave much to be desired. Ion-exchange chromatography was carried out at a flow rate of 1.6 ml/min with a linear salt-gradient made by controlling buffer A and buffer B using a solvent programmer at 20—25°C. The use of high viscosity chromatography buffers lead to a decrease in resolution in gel-permeation chromatography and high column pressure. Ion-exchange liquid chromatography is commonly used to separate and purify proteins. However, this procedure has disadvantages. Considerable time is required to accomplish separation and hence the risk of obtaining denatured proteins and inactive enzymes is increased. Buffers containing the non-ionic detergent, Emulgen 911, have been utilized in solubilization and ion-exchange chromatography of cytochrome P-450.
