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Journal of Separation Science 2010-Aug

Sugar treatment of human lipoprotein particles and their separation by capillary electrophoresis.

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Joanna Witos
Geraldine Cilpa
Gebrenegus Yohannes
Katariina Oörni
Petri T Kovanen
Matti Jauhiainen
Marja-Liisa Riekkola

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Entrapment of lipoprotein particles in the extracellular matrix of the arterial intima is a characteristic feature of the development of atherosclerosis, the disease behind myocardial infarction and stroke. In this study, sugars were exploited in the separation of lipoproteins by CE. Monosaccharides, disaccharides and one sugar alcohol used during ultracentrifugal isolation of lipoproteins prevented the strong and unfavorable adsorption of lipoprotein particles on the capillary wall, allowing their selective separation in uncoated fused silica capillary. The effect of ionic strength of the phosphate BGE solution on the separation at physiological pH was clarified. Asymmetrical flow field-flow fractionation and dynamic light scattering showed that sugars affected the structure of lipoproteins by decreasing their sizes. Although in molecular dynamics simulations, only a 19 amino acid peptide of apolipoprotein B-100 and a 15 amino acid peptide of apolipoprotein E were employed, the results also indicated a decrease in lipoprotein size, supporting the asymmetrical flow field-flow fractionation and dynamic light scattering results.

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