Development of a Procedure for Analysis of High Density Lipoprotein Subclasses

Human serum high density lipoprotein subclasses, HDL2 and HDL3 , were isolated by preparative salt density gradient ultracentrifugation and further analyzed by electrophoresis on a 4 to 15% concentration gradient polyacrylamide gel.The separation of the major classes of serum lipoproteins was achieved after a single ultracentrifugation for 272,000 g at 15°c in a swinging bucket rotor. High resolution concentration gradient gel electrophoresis was found to be particularly suitable for the separation of lipoproteins. The isolation of HDL by precipitation methods, heparin-manganese and dextran-magnesium have also been evaluated in this study, and were found not to completely separate HDL from apo B containing lipoproteins.The combined procedure of salt gradient ultracentrifugation for isolation of serum HDL and concentration gradient polyacrylamide gel electrophoresis analysis of HDL2 and HDL3 was further evaluated with a series of normal and lipemic sera. The results suggested that the new procedures can be adopted by the clinical laboratory for the evaluation of serum HDL subclasses in patients with various cardiovascular diseases.