J. Lipid Res.
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A more recent version of this article appeared on June 1, 2008

Papers In Press, published online ahead of print March 12, 2008
J. Lipid Res., doi:10.1194/jlr.D700044-JLR200
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Submitted on December 17, 2007
Revised on March 4, 2008
Accepted on March 11, 2008

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

Michael S. Yee, Darrell V. Pavitt, Tira Tan, Soundararajan Venkatesan, Ian F. Godsland, William Richmond, and Desmond G. Johnston

Metabolic Medicine, Imperial College, London W2 1NY

Corresponding Author: m.yee{at}imperial.ac.uk

Separation of lipoproteins by traditional sequential, salt density floatation is a prolonged process (~72hr) with variable recovery while iodixanol-based, self-generating density gradients provide a rapid (~4hr) alternative. A novel, 3-layered iodixanol gradient was evaluated for its ability to separate lipoprotein fractions in 63 subjects with varying degrees of dyslipidaemia. 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 CV 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, triglyceride and apolipoproteins were approximately 10% higher with iodixanol compared with sequential floatation. The iodixanol gradient effectively discriminated classic lipoproteins and their sub-fractions and there was evidence for improved resolution of lipoproteins with the iodixanol gradient. LDL particles sub-fractionated by the gradient showed good correlation between density and particle size with small dense LDL <25.5nm 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 sub-fractions, giving important information with regards to LDL phenotype from a single centrifugation step with minimal in-vitro modification of lipoproteins.


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