Abstract
BackgroundHigh-density lipoprotein-cholesterol (HDL-C) is generally measured using several homogeneous assays. We aimed to clarify whether apolipoprotein E-containing HDL (apoE-HDL) subfractions are altered during storage, and if so, whether such changes affect the HDL-C concentration measured using homogeneous assays. MethodsWe stored serum from normolipidemic (n=32) and dyslipidemic (n=17) subjects at 4°C for up to 7days. ApoE-HDL subfractions were analyzed using native 2-dimensional gel (native 2D-gel) electrophoresis. HDL-C concentrations were determined using 2 precipitation and 4homogeneous assays. ResultsNative 2D-gel electrophoresis revealed variously sized apoE-HDL subfractions. After 4h incubation at 37°C, subfractions of smaller particles were converted into larger particles by lecithin:cholesterol acyltransferase (LCAT) activity. After 7days storage at 4°C, the smaller subfractions were decreased in the normolipidemic group, accompanying increases in larger subfractions, whereas changes in the respective subfractions varied among individuals in the dyslipidemic group. HDL-C concentrations were significantly lower after storage at 4°C in all assays, except that using Sekisui Medical's reagent. Therefore, changes in HDL-C concentration and apoE-HDL subfractions were independent of each other. ConclusionApoE-HDL subfractions change during storage, but these changes are not linked to those in HDL-C concentration measured using homogeneous assays.
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