Method for quantitating cholesterol in subfractions of serum lipoproteins separated by gradient gel electrophoresis

M. L. Cheng, C. M. Kammerer, W. F. Lowe, B. Dyke, J. L. VandeBerg

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    Abstract

    Extensive heterogeneity in particle size distribution of serum lipoproteins of baboons was resolved by a procedure that combined Sudan black B prestaining, polyacrylamide gradient gel electrophoresis (GGE), and quantitative densitometry. Each densitometric scan represented a continuous distribution of the relative amount of cholesterol in a serum sample, as a function of the lipoprotein particle size. For analytical purposes, each scan was divided into 12 fractions, representing 12 particle size ranges. The relationship between the estimated cholesterol concentrations in the summed GGE/densitometric fractions corresponding to very low-density lipoproteins (VLDL) + low-density lipoproteins (LDL) and those corresponding to high-density lipoproteins (HDL) and concentrations measured by the heparin-Mn2+ precipitation/enzymatic procedure was linear over a broad range. However, a systematic overestimation of HDL cholesterol concentration and an underestimation of VLDL + LDL cholesterol concentration was apparent. Therefore, correction factors were developed for adjusting the estimates of VLDL + LDL and HDL cholesterol concentrations obtained by the GGE/densitometric method. This analytical method is rapid, repeatable, economical, and useful for genetic and dietary research in which cholesterol concentrations in multiple particle size ranges of lipoproteins must be measured in large numbers of samples. It also is adaptable to immunoblotting procedures for detecting the distribution of specific apolipoproteins among the size-resolved lipoproteins.

    Original languageEnglish (US)
    Pages (from-to)657-681
    Number of pages25
    JournalBiochemical Genetics
    Volume26
    Issue number11
    DOIs
    StatePublished - Dec 1988

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    Keywords

    • gradient gel electrophoresis
    • primates
    • quantitative densitometry
    • serum lipoprotein cholesterol

    ASJC Scopus subject areas

    • Biochemistry
    • Ecology, Evolution, Behavior and Systematics
    • Molecular Biology
    • Genetics

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