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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; Lunec, J

Authors

RJ Bevan

MF Durand

PT Hickenbotham

GD Kitas

PR Patel

HR Griffiths

HL Waller

J Lunec



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.

Citation

Bevan, R., Durand, M., Hickenbotham, P., Kitas, G., Patel, P., Podmore, I., …Lunec, J. (2003). Validation of a novel ELISA for measurement of MDA-LDL in human plasma. Free Radical Biology and Medicine, 35(5), 517-527. https://doi.org/10.1016/S0891-5849%2803%2900359-9

Journal Article Type Article
Publication Date Sep 1, 2003
Deposit Date Aug 8, 2007
Journal Free Radical Biology and Medicine
Print ISSN 0891-5849
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 35
Issue 5
Pages 517-527
DOI https://doi.org/10.1016/S0891-5849%2803%2900359-9
Keywords MDA-LDL, ELISA, antibodies, MDA, F2-isoprostanes, HPLC, GC-MS, plasma, free radicals
Publisher URL http://dx.doi.org/10.1016/S0891-5849(03)00359-9