Lipoprotein separation in a novel iodixanol density gradient, for composition, density, and phenotype analysis

Journal of Lipid Research

Soundararajan Venkatesan¹  Tira Tan¹  Darrell V. Pavitt¹  Ian F. Godsland¹  William Richmond¹  Desmond G. Johnston¹  Michael S. Yee¹ 

[1] Section of Endocrinology and Metabolic Medicine, Faculty of Medicine, Imperial College London, St Mary's Hospital, London W2 1NY, United Kingdom;

关键词: ultracentrifugation;    apolipoprotein B;    very low density lipoprotein;    low density lipoprotein;    high density lipoprotein;    lipoprotein (a);   

DOI  :  

来源: DOAJ

【 摘 要 】

Separation of lipoproteins by traditional sequential salt density floatation is a prolonged process (∼72 h) with variable recovery, whereas iodixanol-based, self-generating density gradients provide a rapid (∼4 h) alternative. A novel, three-layered iodixanol gradient was evaluated for its ability to separate lipoprotein fractions in 63 subjects with varying degrees of dyslipidemia. Lipoprotein cholesterol, triglycerides, and apolipoproteins were measured in 21 successive iodixanol density fractions. Iodixanol fractionation was compared with sequential floatation ultracentrifugation. Iodixanol gradient formation showed a coefficient of variation of 0.29% and total lipid recovery from the gradient of 95.4% for cholesterol and 84.7% for triglyceride. Recoveries for VLDL-, LDL-, and HDL-cholesterol, triglycerides, and apolipoproteins were approximately 10% higher with iodixanol compared with sequential floatation. The iodixanol gradient effectively discriminated classic lipoproteins and their subfractions, and there was evidence for improved resolution of lipoproteins with the iodixanol gradient. LDL particles subfractionated by the gradient showed good correlation between density and particle size with small, dense LDL (<25.5 nm) separated in fractions with density >1.028 g/dl. The new iodixanol density gradient enabled rapid separation with improved resolution and recovery of all lipoproteins and their subfractions, providing important information with regard to LDL phenotype from a single centrifugation step with minimal in-vitro modification of lipoproteins.

【 授权许可】

Unknown