Ion-exchange fast protein liquid chromatography; optimization of the purification of cytochrome P-450 from marmoset monkeys.
Fast protein liquid chromatography (FPLC) on Mono Q and Mono S ion-exchange columns was employed to purify marmoset monkey hepatic cytochrome P-450. Cholate-solubilized liver microsomes from untreated animals as well as from animals induced with phenobarbital, beta-naphthoflavone, 3-methylcholanthrene, and ethanol were used as starting materials. Since established purification schemes for extensively studied species, such as the rat, were found to be not directly applicable, a purification method was developed and optimized by studying the effects of varying detergent types, detergent concentrations, elution buffers, pH values, elution salts, and flow-rates on the resolution and recovery obtained in analytical chromatograms.
D000818:Animals; D002021:Buffers; D002143:Callitrichinae; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification*; D003902:Detergents; D005782:Gels; D006863:Hydrogen-Ion Concentration; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*
D002021:Buffers; D003902:Detergents; D005782:Gels; D003577:Cytochrome P-450 Enzyme System