Comparison of different transition metal ions for immobilized metal affinity chromatography of selenoprotein P from human plasma

摘要

Cu~(2+),Ni~(2+),Zn~(2+),Co~(2+) and Cd~(2+) were evaluated in metal ion affinity chromatography for enrichment of selenoprotein P,and immobilized Co~(2+) affinity chromatography was found to be the most selective chromatographic method. The chromatography was performed by fast protein liquid chromatography and the fractionation was followed by analysis of the collected fractions for selenium by inductively coupled plasma mass spectrometry. By the combination of immobilized Co~(2+) affinity chromatography and heparin affinity chromatography a simple method was developed yielding a 14 800-fold evrichment of selenoprotein P. The purity of the protein was determined by SDS-PAGE and by sequencing from polyvinylidene diflouride blots of SDS-PAGE gels.