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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Simplified method for the diameter sizing of serum low-density lipoprotein using polyacrylamide gradient gel electrophoresis

Hideko Tsukamoto1 / Izumi Takei2 / Keiko Ishii3 / Kiyoaki Watanabe4

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Corresponding author: Hideko Tsukamoto, Department of Laboratory Medicine, Keio University, School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. Phone: 81-3-3353-1211, ext. 62515, Fax: +81-3-3359-6963, E-mail:

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 42, Issue 9, Pages 1009–1012, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2004.204, June 2005

Publication History

Received:
December 9, 2003
Accepted:
August 20, 2004
Published Online:
2005-06-01

Abstract

The appearance of small, dense, low-density lipoprotein in serum has been demonstrated to be associated with increased risk of coronary artery disease. The molecular diameter of low-density lipoprotein is usually measured on the basis of mobility differences on polyacrylamide gel electrophoresis. However, since mobility assessed by this method is seriously affected by the increased levels of serum free fatty acids associated with hypertriglyceridemia, we used polyacrylamide gradient gel electrophoresis to eliminate the interference by fatty acids and devised a simple, precise method of polyacrylamide gradient gel electrophoresis to measure the diameter of small, dense, low-density lipoproteins in serum. We used apoferritin and thyroglobulin, which have a molecular diameter of 12.2 nm and 17.0 nm, respectively, and standard low-density lipoprotein particles having a diameter of 25.7 and 27.0 nm as calibrators, estimated by measurement of negative staining of electron microscopy. We also included apoferritin as an internal standard for polyacrylamide gradient gel electrophoresis. The only stain used was Coomassie brilliant blue, and it was used for lipoprotein staining. When we used low-density lipoprotein of 25.73 nm in diameter as a quality control specimen, the coefficient of variation of the size measurements obtained by our method was less than 1.2%. The new method markedly improved the laboratory procedure for measuring the diameter of low-density lipoproteins.

Keywords: apoferritin; Coomassie brilliant blue; internal standard; measurement of low-density lipoprotein diameter; small, dense, low-density lipoprotein

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