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Journal of Lipid Research, Vol 24, 1060-1069, Copyright © 1983 by Lipid Research, Inc.

ARTICLES

Apolipoprotein E phenotyping with a single gel method: application to the study of informative matings

D Bouthillier, CF Sing and J Davignon

Two-dimensional gel electrophoresis or isoelectric focusing before and after treatment with cysteamine are currently used to determine the six apolipoprotein E isomorphic phenotypes from isolated very low density lipoproteins. A technique is described that makes this possible by performing isoelectric focusing on a single polyacrylamide cylindrical gel under standardized conditions. The technique is simple and accurate enough to obtain 99.5% concordance when the gels are interpreted independently by four different skilled and unskilled observers in the absence of any knowledge of the origin of the samples. There was complete agreement between our technique and the bidimensional method carried out independently in another laboratory on 74 aliquots of plasma very low density lipoproteins. Its application to 16 informative matings involving 101 subjects confirmed the recent demonstration that the apolipoprotein E phenotype inheritance is autosomal and compatible with three common alleles acting at a single genetic locus. Analyses of the contribution of apoE polymorphism to lipid and lipoprotein variability demonstrated a recessive allelic effect of epsilon 2 on plasma very low density lipoprotein cholesterol and a dominant epsilon 4 effect on low density lipoprotein cholesterol. As much as 30% of the variability in low density lipoprotein cholesterol was attributable to this polymorphic gene locus. A simplified scheme is proposed for the symbolic representation of the six phenotypes for clinical and genetic applications.
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