Electrophoretic Separation of LDL and HDL Subclasses

  • David L. Rainwater
Part of the Methods in Molecular Biology™ book series (MIMB, volume 110)

Abstract

An extensive literature supports the hypothesis that measurements of lipoprotein subclasses provide a more detailed reflection of lipoprotein metabolism and a more accurate prediction for risk of cardiovascular disease. Lipoprotein subclasses have been resolved on the basis of density fractionation, gel filtration, and immunological and electrophoretic procedures. Perhaps one of the most widely used methods has been the resolution of lipoproteins on the basis of particle size using nondenaturing pore gradient gel electrophoresis (GGE). GGE procedures have been based on highly reliable gradient gels supplied in two formats: PAA2/16 for resolving low-density lipoproteins (LDLs) and PAA4/30 for resolving high-density lipoprotems (HDLs) (Pharmacia, Piscataway, NJ). These gel formats produce repeatable and detailed separations of the two major classes of lipoprotein particles, appropriate for quantitative analyses of lipoprotein phenotypes.

Keywords

Cholesterol Sucrose Albumin Foam Ethylene Glycol 

References

  1. 1.
    Rainwater, D L, Andres, D. W., Ford, A L, Lowe, W. F, Blanche, P J, and Krauss, R. M. (1992) Production of polyacrylamide gradient gels for the electrophoretic resolution of lipoproteins. J Lipid Res 33, 1876–1881.PubMedGoogle Scholar
  2. 2.
    Li, Z., McNamara, J R, Ordovas, J. M., and Schaefer, E. J. (1994) Analysis of high density lipoproteins by a modified gradient gel electrophoresis method J Lipid Res 35, 1698–1711PubMedGoogle Scholar
  3. 3.
    Singh, ATK, Rainwater, D. L, Haffner, S M, VandeBerg, J. L, Shelledy, W R., Moore, P. H, Jr, and Dyer, T D. (1995) Effect of diabetes on lipoprotein size Arterioscler Thromb. Vasc Biol 15, 1805–1811PubMedGoogle Scholar
  4. 4.
    Schjeide, O A, Rivin, A U, and Yoshmo, J (1963) Uptake of lipid stains by lipids and serum lipoproteins. Am J Clin Patho1 39, 329–341Google Scholar
  5. 5.
    Callais, F, Roche, D., and Andreux, J. P. (1987) Value of polyacrylamide gradient gel electrophoresis of lipoproteins for determining HDL cholesterol Clin Chem 33, 1266.PubMedGoogle Scholar
  6. 6.
    Gambert, P, Fariner, M, Bouzerand, C., Athias, A, and Lallemant, C. (1988) Direct quantitation of serum high density lipoprotein subfractions separated by gradient gel electrophoresis Clin Chim Acta 172, 183–190PubMedCrossRefGoogle Scholar
  7. 7.
    Cheng, M.-L., Kammerer, C. M., Lowe, W. F., Dyke, B, and VandeBerg, J. L. (1988) Method for quantitating cholesterol in subfractions of serum lipoproteins separated by gradient gel electrophoresis. Biochem Genet 26, 657–681.PubMedGoogle Scholar
  8. 8.
    McNamara, J R., Campos, H, Ordovas, J. M, Peterson, J., Wilson, P W F, and Schaefer, E. J (1987) Effect of gender, age, and lipid status on low density lipoprotein subfraction distribution Arteriosclerosis 7, 483–490PubMedGoogle Scholar
  9. 9.
    Rainwater, D. L., Kammerer, C. M., Cheng, M-L, Sparks, M. L., and VandeBerg, J. L. (1992) Distribution of specific apolipoproteins detected by immunoblotting of baboon lipoproteins resolved by polyacrylamide gradtent gel electrophoresis Biochem Genet 30, 143–158.PubMedCrossRefGoogle Scholar
  10. 10.
    Rainwater, D. L., Blangero, J, Moore, P. H., Jr., Shelledy, W R., and Dyer, T. D (1995) Genetic control of apolipoprotein A-I distribution among HDL subclasses Atherosclerosis 118, 307–317.PubMedCrossRefGoogle Scholar
  11. 11.
    Greenwood, F. C., Hunter, W. M., and Glover, J. S (1963) The preparation of 131-labelled human growth hormone of high specific radioactivity Biochem J 89, 114–123PubMedGoogle Scholar
  12. 12.
    Montelaro, R. C. and Salinovich, O (1986) Reversible staining and peptide mapping of protems transferred to nitrocellulose after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis Anal Biochem 156, 341–347PubMedCrossRefGoogle Scholar
  13. 13.
    Blanche, P. J, Gong, E. L., Forte, T. M., and Nichols, A. V (1981) Characterization of human htgh-density lipoproteins by gradient gel electrophoresis Biochim Biophys Acta 665, 408–419PubMedGoogle Scholar
  14. 14.
    Krauss, R. M. and Burke, D J (1982) Identification of multiple subclasses of plasma low density lipoproteins in normal humans J Lipid Res 23, 97–104PubMedGoogle Scholar
  15. 15.
    Lefevre, M. (1988) Localization of lipoprotein unesterified cholesterol in nondenaturing gradient gels with filipin. J Lipid Res 29, 815–818PubMedGoogle Scholar
  16. 16.
    Williams, P. T, Krauss, R. M., Nichols, A. V., Vranizan, K. M., and Wood, P. D S (1990) Identifying the predominant peak diameter of high-density and low-density lipoproteins by electrophoresis. J. Lipid Res 31, 1131–1139PubMedGoogle Scholar
  17. 17.
    Krauss, R. M. and Blanche, P. J. (1992) Detection and quantitation of LDL subfractions Curr Open Lipidol 3, 377–383.CrossRefGoogle Scholar
  18. 18.
    Austin, M A, Brunzell, J D, Fitch, W. L, and Krauss, R M (1990) Inheritance of low density lipoprotein subclass patterns in familial combined hyperlipidemia Arteriosclerosis 10, 520–530.PubMedGoogle Scholar
  19. 19.
    Austin, M A., Pairitz Jarvik, G., Hokanson, J E, and Edwards, K (1993) Complex segregation analysis of LDL peak particle diameter. Genet Epidemiol 10, 599–604PubMedCrossRefGoogle Scholar
  20. 20.
    Campos, H., Blijlevens, E., McNamara, J. R., Ordovas, J M, Posner, B. M., Wilson, P W F, Castelli, W P., and Schaefer, E. J. (1992) LDL particle size distribution: results from the Framingham Offspring Study. Arterioscler Thromb 12, 1410–1419PubMedGoogle Scholar
  21. 21.
    Williams, P. T, Vranizan, K M, and Krauss, R. M. (1992) Correlations of plasma lipoproteins with LDL subfractions by particle size in men and women J Lipid Res. 33, 765–774.PubMedGoogle Scholar
  22. 22.
    Verdery, R. B., Benham, D. F, Baldwin, H. L., Goldberg, A P, and Nichols, A V (1989) Measurement of normative HDL subfraction cholesterol levels by Gaussian summation analysis of gradient gels J, Lipid Res 30, 1085–1095Google Scholar
  23. 23.
    Williams, P T, Krauss, R. M, Vranizan, K. M, Stefanick, M L, Wood, P D. S, and Lindgren, F. T (1992) Associations of lipoproteins and apolipoproteins with gradient gel electrophoresis estimates of high density lipoprotein subfractions in men and women Arterioscler Thromb 12, 332–340.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1998

Authors and Affiliations

  • David L. Rainwater
    • 1
  1. 1.Department of GeneticsSouthwest Foundation for Biomedical ResearchSan Antonio

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