Purification of complex protein mixtures by ion-exchange displacement chromatography using spacer displacers.
The anion-exchange separation of complex protein mixtures by displacement chromatography using spacer displacers driven by high-affinity final displacers is demonstrated. Guinea pig serum was separated on a medium-resolution adsorbent using a single heterogeneous mixture of carboxymethyldextran displacers to space the protein components. Mouse liver cytosol was separated on a low-resolution adsorbent using six carboxymethyldextran spacer displacers of increasing column affinity. The demonstration of the purification of alkaline phosphatase from E. coli periplasm by displacement chromatography on a high-performance liquid chromatography column is reviewed. The benefits of spacer displacers for separating minor components from complex biological protein mixtures is discussed. A simplified method for preparing carboxymethyldextran displacers is presented.
Torres
A R
AR
Bio-Fractionations, Logan, UT 84321.
Peterson
E A
EA
D000327:Adsorption; D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification; D001798:Blood Proteins / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003911:Dextrans; D004926:Escherichia coli / Q000737:chemistry; D006168:Guinea Pigs; D008099:Liver / Q000737:chemistry; D051379:Mice; D011506:Proteins / Q000302:isolation & purification*
D001426:Bacterial Proteins; D001798:Blood Proteins; D003911:Dextrans; D011506:Proteins; C014392:carboxymethyl dextran