Isolation of lipoprotein (a) using the regenerate of a dextran sulfate cellulose LDL apheresis system.
A simple method for the preparation of lipoprotein (a) is presented. The procedure uses the eluate of an LDL apheresis system operating on the basis of LDL adsorbing dextran sulfate cellulose. The eluate is concentrated by tangential flow membrane filtration and subjected to ultracentrifugation, first at a density of 1.125 kg/liter and then at 1.050 kg/liter. The crude lipoprotein (a)-containing fraction is chromatographed on agarose (Bio-Gel A-15m) to remove contaminating low-density and high-density lipoproteins. As demonstrated by immunoelectrophoresis with intermediate gel, the method provides lipoprotein (a) completely free of LDL. SDS-polyacrylamide gel electrophoresis showed that apolipoprotein E was associated with purified lipoprotein (a). On agarose gel electrophoresis and two-dimensional immunoelectrophoresis, lipoprotein (a) prepared by the proposed method cannot be distinguished from native lipoprotein (a). The major advantage of the procedure is that it allows the isolation of large amounts of lipoprotein (a) from a single donor.
Gross
E
E
Center of Internal Medicine, Johann Wolfgang Goethe-University, Frankfurt/Main, Federal Republic of Germany.
März
W
W
Siekmeier
R
R
Scharrer
I
I
Gross
W
W
D000327:Adsorption; D001781:Blood Component Removal; D002482:Cellulose; D002850:Chromatography, Gel; D016264:Dextran Sulfate; D004587:Electrophoresis, Agar Gel; D006493:Heparin; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000097:blood; D007122:Immunoelectrophoresis; D017270:Lipoprotein(a) / Q000302:isolation & purification*; D008077:Lipoproteins, LDL / Q000737:chemistry* / Q000302:isolation & purification; D014461:Ultracentrifugation
D017270:Lipoprotein(a); D008077:Lipoproteins, LDL; D002482:Cellulose; D006493:Heparin; D016264:Dextran Sulfate