Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter June 1, 2005

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

  • Hideko Tsukamoto , Izumi Takei , Keiko Ishii and Kiyoaki Watanabe

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.


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:

References

1 Krauss RM, Burke DJ. Identification of multiple subclasses of plasma low-density lipoproteins in normal humans. J Lipid Res 1982; 23:97–104.10.1016/S0022-2275(20)38178-5Search in Google Scholar

2 Sacks FM, Campos H. Cardiovascular endocrinology 4. Low-density lipoprotein size and cardiovascular disease: a reappraisal. J Clin Endocrinol Metab 2003; 88:4525–532.10.1210/jc.2003-030636Search in Google Scholar

3 Kuller L, Arnold A, Tracy R, Otvos J, Burke G, Psaty B, et al. Nuclear magnetic resonance spectroscopy of lipoproteins and risk of coronary heart disease in the cardiovascular health study. Arterioscler Thromb Vasc Biol 2002; 22:1175–80.10.1161/01.ATV.0000022015.97341.3ASearch in Google Scholar

4 Williams PT, Krauss RM, Nochols AV, Vranizan KM, Wood PDS. Identifying the predominant peak diameter of high-density and low-density lipoproteins by electrophoresis. J Lipid Res 1990; 31:1131–9.10.1016/S0022-2275(20)42753-1Search in Google Scholar

5 Kondo A, Muranaka Y, Ohta I, Kanno T. Dynamic reaction in a homogeneous HDL-cholesterol assay visualized by electron microscopy. Clin Chem 1999; 45:1974–80.10.1093/clinchem/45.11.1974Search in Google Scholar

Received: 2003-12-9
Accepted: 2004-8-20
Published Online: 2005-6-1
Published in Print: 2004-9-1

© Walter de Gruyter

Downloaded on 5.3.2024 from https://www.degruyter.com/document/doi/10.1515/CCLM.2004.204/html
Scroll to top button