Anion-exchange fast protein liquid chromatographic characterization and purification of apolipoproteins A-I, A-II, C-I, C-II, C-III0, C-III1, C-III2 and E from human plasma.
This paper describes a procedure for the rapid isolation of urea-soluble apolipoproteins (apo) from delipidated human very-low- and high-density lipoproteins using anion-exchange fast protein liquid chromatography. The separation was complete within 30 min and peaks corresponding to apolipoproteins A-I, A-II, C-I, C-II, C-III0, C-III1, C-III2 and E were identified by comparing their chromatographic, electrophoretic and immunological behaviour with that of purified standards of each protein. A second purification step is necessary to obtain pure apolipoproteins. Apo E, which is difficult to purify by conventional chromatography, has been obtained in a good yield. The apo C-II that was obtained produced a symmetrical peak on chromatography but three bands in isoelectric focusing. The method can be upgraded to a preparative scale and offers the possibility of direct purification of apolipoproteins both from high-density lipoproteins and (following preliminary gel chromatography) from very-low-density lipoproteins.
D000838:Anions; D016632:Apolipoprotein A-I; D016633:Apolipoprotein A-II; D053302:Apolipoprotein C-I; D053304:Apolipoprotein C-II; D053305:Apolipoprotein C-III; D001054:Apolipoproteins A / Q000097:blood; D001056:Apolipoproteins C / Q000097:blood; D001057:Apolipoproteins E / Q000097:blood; D001059:Apoproteins / Q000097:blood*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006801:Humans; D007124:Immunoenzyme Techniques
D000838:Anions; D016632:Apolipoprotein A-I; D016633:Apolipoprotein A-II; D053302:Apolipoprotein C-I; D053304:Apolipoprotein C-II; D053305:Apolipoprotein C-III; D001054:Apolipoproteins A; D001056:Apolipoproteins C; D001057:Apolipoproteins E; D001059:Apoproteins