CHROMATOGRAPHIC-SEPARATION AND BEHAVIOR OF MICROSOMAL CYTOCHROME-P450AND CYTOCHROME B(5)

The methods used for separation of the multiple mammalian cytochrome P 450 enzymes by liquid chromatography are reviewed. In addition to the chromatographic techniques, preparation and handling of samples and pr efractionation procedures are considered. Conditions that affect stabi lity and chromatographic resolution of cytochromes P450 are also discu ssed. Special emphasis is put on useful methods which are not routinel y used for P450 separation, such as immobilized metal affinity or hydr ophobic-interaction chromatography. Applications of low- and high-pres sure methods with regard to preparative and analytical separations are compared. It is shown that high- and medium-pressure ion-exchange chr omatography are suitable tools for separation of closely related P450 enzymes, especially when specific detection methods are available. In addition to fractionation of cytochromes P450, the isolation and chrom atographic behavior of cytochrome b(5) is discussed.