Abstract
The selective and reversible adsorption of bovine low density lipoproteins (LDL) by heparin-Sepharose has been exploited as the critical step in a procedure for the preparative isolation of very low density lipoproteins (VLDL)/chylomicrons, LDL, and high density lipoproteins (HDL) from bovine plasma. Molecular size exclusion chromatography and isopycnic density gradient separation steps are also involved in the method described. The resulting HDL and LDL fractions are free from contamination by one another as judged by electrophoretic mobility in agarose gels. The major lipid and apolipoprotein compositions of the three resolved lipoprotein classes have been determined.
Highlights
Abrtract The selective and reversible adsorption of bovine low density lipoproteins (LDL) by heparin-Sepharose has been exploited as the critical step in a pmcdure for the preparative isolation of very low density lipoproteins (VLDL)/chylomicrons, LDL, and high density lipoproteins (HDL) from &ne plasma
Flotation of lipoproteins contained in peak B fractions in buoyant density gradients of KBr revealed the presence of a single peak of protein centered on a density of 1.08 g/ml
Agarose gel electrophoretic analysis indicated the presence of both a-and 8-migrating lipoproteins in peak B, suggesting the presence of both LDL and HDL. By this technique, of fractions across the density gradient revealed considerable overlap in the buoyant density characteristics of the two electrophoretically discernible lipoproteins; the equilibrium density flotation method was unsuitable for the quantitative separation of LDL from HDL, as previously found by others [5,6,7, 10, 20, 21]
Summary
Abrtract The selective and reversible adsorption of bovine low density lipoproteins (LDL) by heparin-Sepharose has been exploited as the critical step in a pmcdure for the preparative isolation of very low density lipoproteins (VLDL)/chylomicrons, LDL, and high density lipoproteins (HDL) from &ne plasma. Molecular size exclusion chromatography and isopycnic density gradient separation steps are involved in the method described. The resulting HDL and LDL fractions arc f m from contamination by one another as judged by electrophoretic mobility in agarose gels. The major lipid and apolipoprotein compositions of the three resolved lipoprotein classes have been dettrmined.-Cordle, S. Purification and characterization of bovine lipoproteins: resolution of high density and low density lipoproteins using heparinSepharose chromatography.J Lipid Rar. 1985. Purification and characterization of bovine lipoproteins: resolution of high density and low density lipoproteins using heparinSepharose chromatography.J Lipid Rar. 1985. 26: 721-725
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