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Journal of Lipid Research, Vol. 44, 212-217, January 2003
Copyright © 2003 by Lipid Research, Inc.

Methods

Non-denaturing polyacrylamide gradient gel electrophoresis for the diagnosis of dysbetalipoproteinemia

Dirk J. Blom, Pamela Byrnes, Sheena Jones and A. David Marais¹

Lipidology Division of the Department of Medicine and MRC Cape Heart Group, University of Cape Town, Faculty of Health Sciences, Observatory, South Africa

¹ To whom correspondence should be addressed. e-mail: dmarais{at}capeheart.uct.ac.za

Dysbetalipoproteinemia, an uncommon but highly atherogenic mixed hyperlipidemia due to the accumulation of remnants of triglyceride-rich lipoproteins, is characterized by cholesterol-enriched VLDL that migrates in the ß-position on agarose gels. The demonstration of a broad ß-band on agarose gel electrophoresis of plasma is an insensitive method and ultracentrifugation is an impractical method of diagnosing this condition. Non-denaturing polyacrylamide gradient gel electrophoresis (PGGE) was investigated as a screening method for the diagnosis of dysbetalipoproteinemia. A minigel procedure separating the Sudan Black prestained apolipoprotein B (apoB)-containing lipoproteins on a 2–8% polyacrylamide gel at 4°C overnight was analyzed for ultracentrifugally and genetically proven dysbetalipoproteinemic subjects as well as matched controls for mixed hyperlipidemia. Visual inspection revealed that the presence of only small VLDL- and IDL-like particles in untreated patients was highly sensitive (72%) and specific (95%) for dysbetalipoproteinemia. Videodensitometric analysis of area under the curve for large and small VLDL, as well as IDL and LDL, permitted even better discrimination in subjects whose profiles included some staining in the LDL-like region. A ratio of area under the curve of more than 0.5 for IDL-LDL allowed for a specificity of 100% and a sensitivity of 89% for the diagnosis of dysbetalipoproteinemia.

This modified PGGE system may be useful in screening for dysbetalipoproteinemia.

Abbreviations: HSPG, heparin sulfate proteoglycans; PGGE, polyacrylamide gradient gel electrophoresis; R_f, retardation factor; TEMED, tetramethylethylenediamine; TGRL, triglyceride-rich lipoproteins

Supplementary key words Type III hyperlipidemia • remnants • apolipoprotein E • method

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This article has been cited by other articles:

				D. J. Blom, F. H. O'Neill, and A. D. Marais Screening for Dysbetalipoproteinemia by Plasma Cholesterol and Apolipoprotein B Concentrations Clin. Chem., May 1, 2005; 51(5): 904 - 907. [Full Text] [PDF]