Lipoprotein Agarose Gel Electrophoresis. Application in HDL-Cholesterol Methodology.
Interim rept. Jan 80-Jan 85,
SCHOOL OF AEROSPACE MEDICINE BROOKS AFB TX
Pagination or Media Count:
We perform agarose gel lipoprotein electrophoresis AGLE in this laboratory using glass microscope slides to support the agarose gel and 0.025 M barbital buffer for the electrophoresis. This report describes details, including special apparatus used to facilitate making and handling agarose gel slides. To test correlations between serum lipid levels and the densities of the corresponding lipoprotein bands separated by electrophoresis, we electrophoresed eight individual sera and measured the densities of the resulting lipoprotein bands. Serum level of total cholesterol, triglycerides, and HDL-cholesterol were correlated with the measured densities of the beta, prebeta, and alpha lipoprotein bands respectively. Coefficients for correlation between total cholesterol and beta lipoprotein, triglyceride and prebeta lipoprotein, and HDL-cholesterol and alpha lipoprotein were 0.638, 0.914, and 0.920 respectively. Densitometric data from 24 replicate electrophoresis slides showed that the coefficient of variance for the percentage of dye bound to the beta, prebeta, and alpha lipoproteins was 1.9, 2.6 and 4.5 respectively. We used AGLE to monitor the separation of high-density lipoprotein HDL from the combined low-density lipoproteins LDL and very low density lipoproteins VLDL during the titration of serum lipoproteins with increasing amounts of lipoprotein precipitant. Results demonstrated which levels of precipitant removed all detectable VLDL LDL and which levels of precipitant removed a significant amount of HDL. These data, together with the levels of cholesterol in the supernate from the various tubes, identified the optimum precipitant volumes to be used in routine determinations of serum HDL-cholesterol levels.