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

Ruth J. Bevan*, Mireille F. Durand, Peter T. Hickenbotham, George D. Kitas, Parul R. Patel, Ian D. Podmore, Helen R. Griffiths, Helen L. Waller, Joseph Lunec

*Corresponding author for this work

Research output: Contribution to journalArticle

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 μg/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 μg/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. © 2003 Elsevier Inc.

Original languageEnglish
Pages (from-to)517-527
Number of pages11
JournalFree Radical Biology and Medicine
Volume35
Issue number5
DOIs
Publication statusPublished - 1 Sep 2003

Keywords

  • antibodies
  • ELISA
  • f-isoprostanes
  • free radicals
  • GC-MS
  • HPLC
  • MDA
  • MDA-LDL
  • plasma

Fingerprint Dive into the research topics of 'Validation of a novel ELISA for measurement of MDA-LDL in human plasma'. Together they form a unique fingerprint.

  • Cite this

    Bevan, R. J., Durand, M. F., Hickenbotham, P. T., Kitas, G. D., Patel, P. R., Podmore, I. D., Griffiths, H. R., Waller, H. L., & 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(03)00359-9