Article on Methodology

Analytical capillary isotachophoresis of total plasma lipoproteins: a new tool to identify atherogenic low density lipoproteins

Correspondence to: Gabriele Bittolo-Bon

Plasma low density lipoproteins from 20 patients were separated by capillary isotachophoresis (ITP). In each patient the apparent diameter of the predominant LDL peak on whole plasma was also determined by nondenaturing gradient gel electrophoresis. Furthermore the concentration of the more electronegatively charged in vivo oxidized LDL^- was accomplished using anion exchange high pressure liquid chromatography. By analytical capillary ITP of whole plasma lipoproteins, prestained with a lipophilic dye, LDL were separated into four subfractions. Usually, the predominant subfraction was the slow migrating LDL₄, followed by LDL₃, and then by the faster LDL₂ and LDL₁. Slow migrating LDL₄ correlated negatively with plasma triglycerides and LDL^- and positively with plasma high density lipoprotein (HDL) cholesterol and with the LDL diameter, while the faster LDL₁ showed an inverse behavior. The LDL₁ + LDL₂ to LDL₃ + LDL₄ ratio showed a strong positive correlation with LDL^- concentration (r = 0.87; P < 0.001) and a highly significant inverse correlation with the LDL particle diameter (r = -0.74; P < 0.001). At least three highly atherogenic LDL that could be found in human plasma, namely oxidized, glycated and small-dense, are characterized by a greater electric charge. The LDL profile from capillary ITP and the relative prevalence of faster or slower migrating LDL fractions could indicate the presence of more atherogenic LDL.—Bittolo-Bon, G., and G. Cazzolato. Analytical capillary isotachophoresis of total plasma lipoproteins: a new tool to identify atherogenic low density lipoproteins. J. Lipid Res. 1999. 40: 170–177.

Supplementary key words: oxidized LDL, LDL size, lipoprotein subfractionation

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