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A quick and large-scale density gradient subfractionation method for low density lipoproteins.

D M Lee, D Downs

    Journal of Lipid Research
    |January 1, 1982
    PubMed
    Summary
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    A new method quickly separates lipoproteins from plasma using density gradient ultracentrifugation. This technique reveals compositional differences between males and females and aids in studying dyslipoproteinemias.

    Area of Science:

    • Biochemistry
    • Lipidology
    • Analytical Chemistry

    Background:

    • Lipoproteins are crucial for lipid transport; their subclasses have distinct roles in health and disease.
    • Existing methods for lipoprotein subfractionation can be time-consuming and may not yield sufficiently pure fractions.
    • Understanding lipoprotein heterogeneity is key to diagnosing and managing dyslipoproteinemias.

    Purpose of the Study:

    • To develop a rapid, large-scale density gradient subfractionation method for lipoproteins with density < 1.063 g/ml.
    • To characterize the subfractions obtained from normal male and female plasma.
    • To assess the method's utility in analyzing dyslipoproteinemias.

    Main Methods:

    • Developed a single ultracentrifugation step using a discontinuous density gradient (plasma over 1.080 g/ml NaCl solution).

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  • Separated up to 324 ml of plasma into five distinct layers based on density.
  • Verified the method's reproducibility and accuracy using analytical ultracentrifugation, chemical analyses, electrophoresis, and electron microscopy.

    • Main Results:

    • Successfully resolved VLDL, LDL(1), LDL(2) subfractions (LDL2a, LDL2b, LDL2c), HDL, and plasma proteins.
    • Identified significant compositional differences in triglyceride, cholesteryl ester, and cholesterol concentrations between normal males and females.
    • Demonstrated distinct lipoprotein profiles in patients with type III and type IV hyperlipoproteinemias compared to normal subjects.

    Conclusions:

    • The developed method provides a quick and reproducible way to isolate concentrated lipoprotein subfractions, including heterogeneous LDL(2) species.
    • The technique is valuable for identifying sex-specific lipoprotein compositional differences.
    • This subfractionation approach is applicable to the study of various dyslipoproteinemias, offering insights into disease-specific alterations.