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Journal of Lipid Research, Vol. 45, 1572-1579, August 2004
Copyright © 2004 by American Society for Biochemistry and Molecular Biology

Methods

Storage of human plasma samples leads to alterations in the lipoprotein distribution of apoC-III and apoE

Jeffrey S. Cohn¹, Claudia Rodriguez, Hélène Jacques, Michel Tremblay and Jean Davignon

Clinical Research Institute of Montreal, Quebec, Canada

¹ To whom correspondence should be addressed. e-mail: cohnj{at}ircm.qc.ca

The effect of frozen storage on lipoprotein distribution of apolipoprotein C-III (apoC-III) and apoE was investigated by measuring apoC-III and apoE by ELISA in HDL and apoB-containing lipoproteins of human plasma samples (n = 16) before and after 2 weeks of frozen storage (–20°C). HDLs were separated by heparin-manganese precipitation (HMP) or by fast-protein liquid chromatography (FPLC). Total plasma apoC-III and apoE levels were not affected by frozen storage. HDL-HMP apoC-III and apoE levels were significantly higher in frozen versus fresh samples: 7.7 ± 0.7 versus 6.7 ± 0.7 mg/dl (P < 0.05) and 2.0 ± 0.1 versus 1.2 ± 0.1 mg/dl (P < 0.001), respectively. HDL-FPLC apoC-III and apoE, but not triglyceride (TG) or cholesterol, levels were also higher in frozen samples: 12.0 ± 1.2 versus 7.5 ± 0.6 mg/dl (P < 0.001) and 2.7 ± 0.2 versus 1.6 ± 0.2 mg/dl (P < 0.001), respectively. Frozen storage led to a decrease in apoC-III (–17 ± 9%) and apoE (–19 ± 9%) in triglyceride-rich lipoprotein. Redistribution of apoC-III and apoE was most evident in samples with high TG levels. HDL apoC-III and apoE levels were also significantly higher when measured in plasma stored at –80°C.

Our results demonstrate that lipoprotein distribution of apoC-III and apoE is affected by storage of human plasma, suggesting that analysis of frozen plasma should be avoided in studies relating lipoprotein levels of apoC-III and/or apoE to the incidence of coronary artery disease.

Abbreviations: apo, apolipoprotein; CAD, coronary artery disease; FPLC, fast-protein liquid chromatography; HC, hypercholesterolemic; HMP, heparin-manganese precipitation; HTG, hypertriglyceridemic; NL, normolipidemic; TG, triglyceride; TRL, triglyceride-rich lipoprotein

Supplementary key words apolipoprotein • atherosclerosis • cholesterol • coronary artery disease • HDL • remnants • triglyceride

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