Lipoprotein Analysis Using Agarose Gel Electrophoresis and Differential Staining of Lipids

• Published in: Journal of Atherosclerosis and Thrombosis Vol. 8; no. 1; pp. 7 - 13
• Main Authors: Kido, Toshimi, Kurata, Hideaki, Matsumoto, Akiyo, Tobiyama, Rie, Musha, Toshiaki, Hayashi, Katsuji, Tamai, Seiichi, Utsunomiya, Kazunori, Tajima, Naoko, Fidge, Noel, Itakura, Hiroshige, Kondo, Kazuo
• Format: Journal Article
• Language: English
• Published: Japan Japan Atherosclerosis Society 2001
• Subjects: Cholesterol; Chylomicrons - blood; Elecrtophoretic profiling; Electrophoresis, Agar Gel - methods; Humans; Hyperlipidemias - classification; Hyperlipidemias - diagnosis; Hyperlipoproteinemia; Image Processing, Computer-Assisted; Linear Models; Lipids - analysis; Lipoproteins - analysis; Lipoproteins - blood; Software; Staining and Labeling - methods; Triglyceride; Ultracentrifugation - methods
• ISSN: 1340-3478; 1880-3873
• DOI: 10.5551/jat1994.8.7

We established a strategy to directly measure cholesterol and triglyceride levels of each lipoprotein fraction using a combination of agarose gel electrophoresis and differential staining. The cholesterol and triglyceride levels determined by electrophoresis correlated significantly with those of ultracentrifugation. The correlation coefficients between these methods were, for cholesterol levels 0.975 (very low density lipoproteins, VLDL), 0.986 (low density lipoproteins, LDL) and 0.965 (high density lipoproteins, HDL) and for triglyceride levels 0.994 (VLDL), 0.963 (LDL) and 0.959 (HDL) respectively. Both intra-and inter-assays showed low values of coefficients of variation (CV) (less than 3.57%). We observed a strong linearity between staining and triglyceride concentration. An increased VLDL-cholesterol was observed in type Ill subjects, a result which enabled distinction between type Ill and type llb or type V lipoproteinemia. The method revealed lipoprotein patterns in some samples otherwise unexpected from their corresponding serum lipid parameters. Analyses of these electrophoretic patterns thus provide an effective technique to classify types of hyperlipidemia defined by the WHO. Furthermore, quantitative measurement of chylomicrons, usually difficult, proved to be achievable, providing an additional analysis of postprandial hyperlipidemia and the exact measurement of LDL-cholesterol after diet. Consequently, we recommend this simple and easy method for clinical evaluation of abnormalities in lipoprotein profiles.