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Journal of Lipid Research, Vol 23, 1206-1223, Copyright © 1982 by Lipid Research, Inc.
LI Gidez, GJ Miller, M Burstein, S Slagle and HA Eder
Studies in recent years have suggested that measurement of high density
lipoprotein (HDL) subclasses may provide significant information beyond
that provided by measurement of total HDL. However, conventional
methodology for separation of HDL subclasses involves various types of
ultracentrifugation that are time-consuming, costly, and not suitable for
many clinical or epidemiological studies. We have developed a simple
precipitation method for the separation of HDL subclasses in human plasma.
After precipitation of apoB-containing lipoproteins with heparin-Mn2+, HDL2
is precipitated by addition of dextran sulfate (mol wt 15,000). HDL2
cholesterol is calculated as the difference between total HDL cholesterol
(heparin-Mn2+ supernatant) and HDL3 cholesterol (dextran sulfate
supernatant). HDL2 determined by this method correlated well with results
obtained by preparative ultracentrifugation (n = 295, r = 0.91) and
analytical ultracentrifugation (n = 17, r = 0.92). In the original method
final concentration of dextran sulfate was 0.09 g/dl; however further
studies indicated that 0.13 g/dl is a more suitable concentration. The
chemical compositions of HDL2 and HDL3 isolated by the precipitation method
were very similar to those of HDL2 and HDL3 isolated by preparative
ultracentrifugation. The concentration of HDL2 cholesterol was 40% higher
in normal women than in normal men. In men with coronary heart disease,
total HDL was decreased by 28%, HDL2 was decreased by 44%, while HDL3 was
19% lower. A similar pattern of change was found in women with coronary
heart disease. In other conditions where total HDL either increased or
decreased, the change in HDL2 was always proportionately greater than the
change in total HDL. HDL3 showed relatively less change, and in some
instances its concentration was unchanged. Thus HDL2 is the more variable
component and may be a more meaningful index of altered HDL metabolism.
ARTICLES
Separation and quantitation of subclasses of human plasma high density lipoproteins by a simple precipitation procedure
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