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Validation of a novel ELISA for measurement of MDA-LDL in human plasma

Bevan, RJ, Durand, MF, Hickenbotham, PT, Kitas, GD, Patel, PR, Podmore, I, Griffiths, HR, Waller, HL and Lunec, J 2003, 'Validation of a novel ELISA for measurement of MDA-LDL in human plasma' , Free Radical Biology and Medicine, 35 (5) , pp. 517-527.

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Abstract

The involvement of oxidatively modified low density lipoprotein (LDL) in the development of CHD is widely described. We have produced two antibodies, recognizing the lipid oxidation product malondialdehyde (MDA) on whole LDL or ApoB-100. The antibodies were utilized in the development of an ELISA for quantitation of MDA-LDL in human plasma. Intra- and inter-assay coefficients of variation (% CV) were measured as 4.8 and 7.7%, respectively, and sensitivity of the assay as 0.04 mug/ml MDA-LDL. Recovery of standard MDA-LDL from native LDL was 102%, indicating the ELISA to be specific with no interference from other biomolecules. Further validation of the ELISA was carried out against two established methods for measurement of lipid peroxidation products, MDA by HPLC and F2-isoprostanes by GC-MS. Results indicated that MDA-LDL is formed at a later stage of oxidation than either MDA or F2-isoprostanes. In vivo analysis demonstrated that the ELISA was able to determine steady-state concentrations of plasma MDA-LDL (an end marker of lipid peroxidation). A reference range of 34.3 +/- 8.8 mug/ml MDA-LDL was established for healthy individuals. Further, the ELISA was used to show significantly increased plasma MDA-LDL levels in subjects with confirmed ischemic heart disease, and could therefore possibly be of benefit as a diagnostic tool for assessing CHD risk.

Item Type: Article
Uncontrolled Keywords: MDA-LDL, ELISA, antibodies, MDA, F2-isoprostanes, HPLC, GC-MS, plasma, free radicals
Themes: Subjects / Themes > Q Science > Q Science (General)
Subjects / Themes > R Medicine > R Medicine (General)
Health and Wellbeing
Subjects outside of the University Themes
Schools: Colleges and Schools > College of Science & Technology
Colleges and Schools > College of Science & Technology > School of Environment and Life Sciences
Colleges and Schools > College of Science & Technology > School of Environment and Life Sciences > Biomedical Research Centre
Journal or Publication Title: Free Radical Biology and Medicine
Publisher: Elsevier
Refereed: Yes
ISSN: 08915849
Depositing User: H Kenna
Date Deposited: 08 Aug 2007 12:29
Last Modified: 20 Aug 2013 16:45
URI: http://usir.salford.ac.uk/id/eprint/175

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