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Separation of the principal HDL subclasses using iodixanol ultracentrifugation

Harman, NL and Griffin, BA and Davies, IG (2013) Separation of the principal HDL subclasses using iodixanol ultracentrifugation. Journal of Lipid Research, 54. pp. 2273-2281. ISSN 1539-7262

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Abstract

HDL subclasses detection, in cardiovascular risk, has been limited due to the time-consuming nature of current techniques. We have developed a time-saving and reliable separation of the principal HDL subclasses employing iodixanol density gradient ultracentrifugation (IxDGUC) combined with digital photography. HDL subclasses were separated in 2.5 h from prestained plasma on a three-step iodixanol gradient. HDL subclass profiles were generated by digital photography and gel scan software. Plasma samples (n = 46) were used to optimize the gradient for the resolution of HDL heterogeneity and to compare profiles generated by IxDGUC with gradient gel electrophoresis (GGE); further characterization from participants (n = 548) with a range of lipid profiles was also performed. HDL subclass profiles generated by IxDGUC were comparable to those separated by GGE as indicated by a significant association between areas under the curve for both HDL2 and HDL3 (HDL2, r = 0.896, P < 0.01; HDL3, r = 0.894, P < 0.01). The method was highly reproducible, with intra- and interassay coefficient of variation percentage < 5 for percentage area under the curve HDL2 and HDL3, and < 1% for peak Rf and peak density. The method provides time-saving and cost-effective detection and preparation of the principal HDL subclasses.

Item Type: Article
Uncontrolled Keywords: 0601 Biochemistry And Cell Biology, 1101 Medical Biochemistry And Metabolomics
Subjects: Q Science > QD Chemistry
Q Science > QH Natural history > QH301 Biology
Divisions: School of Sport Studies, Leisure and Nutrition
Publisher: American Society for Biochemsitry and Molecular Biology
Date Deposited: 11 Mar 2016 09:00
Last Modified: 11 Mar 2016 09:00
DOI or Identification number: 10.1194/jlr.D037432
URI: http://researchonline.ljmu.ac.uk/id/eprint/3152

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