Rapid ion-exchange chromatography for preparative separation of proteins. Application to porcine and bovine carbonic anhydrases.
A method of rapid ion-exchange chromatography of DEAE-cellulose for preparative purposes is described. Basically, the flow-rate is increased by applying an air pressure on the column. By this technique it is possible to purify gram quantities of protein in 2-4 h with acceptable resolution. In preparations of bovine and porcine carbonic anhydrases the elution times were reduced by a factor of about ten compared to those of conventional methods. The enzymes purified in this way showed a high degree of homogeneity. The method should be generally applicable in protein purification, and especially advantageous in purification of unstable proteins where time-consuming separations often give rise to low yields of active material.
D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000302:isolation & purification*; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology; D011506:Proteins / Q000302:isolation & purification*; D013045:Species Specificity; D013552:Swine
D011506:Proteins; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases