The Effects of Heparin on Lipoproteins in High-Resolution Electrophoresis of Serum

Abstract

Sera from heparinized patients commonly display anodal slurring of both the alpha and beta lipoproteins when they are examined by high-resolution electrophoresis (HRE). In this study, the authors examined the effect of heparin and lipoprotein lipase on the electrophoretic migration of alpha and beta lipoproteins in sera. The addition of 10 to 1,000 units of heparin/mL to normal sera resulted in a concentration-dependent anodal slurring of the beta lipoproteins. At 40 units/mL, the beta lipoprotein band was not visible when the Paragon blue protein stain was used. The beta lipoprotein could be seen as a wide, faintly staining band with lipoprotein stain. The alpha lipoprotein band on the same gel was unaffected by the added heparin. High-resolution electrophoresis of other sera from patients who were therapeutically heparinized demonstrated anodal slurring of both alpha and beta lipoproteins independent of heparin concentration. Immunofixation electrophoresis (IFE) studies confirmed that apolipoproteins A and B were slurred within their respective bands. Heparin activates lipoprotein lipase with release of free fatty acids (FFA) from very low density lipoproteins and chylomicrons. To test this effect on migration, sera were incubated with lipoprotein lipase in vitro. The anodal slurring of both the alpha and beta lipoprotein was associated with the amount of FFA production. Individuals interpreting electrophoretic patterns should be aware that both the alpha and beta lipoproteins can migrate and slur anodally in heparinized patients. In addition, when the beta lipoprotein band interferes with the identification of monoclonal gammopathies, its migration can be selectively altered by the addition of 40 units/mL heparin to the sample.