Purification and characterization of apolipoprotein C-II from human plasma by high-pressure liquid chromatography.
Apolipoprotein C-II, which activates lipoprotein lipase, was isolated from normal subjects and purified to homogeneity by reverse-phase high-pressure liquid chromatography (HPLC). The partially purified product from DEAE-cellulose chromatography was eluted from a Radial Pak C18 cartridge in a radial compression module using a linear gradient of 0.01 M ammonium bicarbonate and acetonitrile. The final product was homogeneous by polyacrylamide gel electrophoresis (pH 8.9), isoelectric focusing (pH 2.5-6.5), Ouchterlony double immunodiffusion, analytical HPLC and amino acid analysis. The purification of apolipoprotein C-II from normal subjects will permit the elucidation of its amino acid sequence and subsequent comparison with the known sequence of apolipoprotein C-II isolated from patients with hyperlipoproteinemia.
D000596:Amino Acids / Q000032:analysis; D053304:Apolipoprotein C-II; D001053:Apolipoproteins / Q000097:blood* / Q000302:isolation & purification; D001056:Apolipoproteins C; D002851:Chromatography, High Pressure Liquid; D006801:Humans
D000596:Amino Acids; D053304:Apolipoprotein C-II; D001053:Apolipoproteins; D001056:Apolipoproteins C