A biochemical fluorometric method for assessing the oxidative properties of HDL[S]

Theodoros Kelesidis; Judith S. Currier; Diana Huynh; David Meriwether; Christina Charles-Schoeman; Srinivasa T. Reddy; Alan M. Fogelman; Mohamad Navab; Otto O. Yang

Journal of Lipid Research (Dec 2011)

A biochemical fluorometric method for assessing the oxidative properties of HDL[S]

Theodoros Kelesidis,
Judith S. Currier,
Diana Huynh,
David Meriwether,
Christina Charles-Schoeman,
Srinivasa T. Reddy,
Alan M. Fogelman,
Mohamad Navab,
Otto O. Yang

Affiliations

Theodoros Kelesidis: To whom correspondence should be addressed.; Department of Medicine, University of California, Los Angeles, CA 90095; To whom correspondence should be addressed.
Judith S. Currier: Department of Medicine, University of California, Los Angeles, CA 90095
Diana Huynh: Department of Medicine, University of California, Los Angeles, CA 90095
David Meriwether: Department of Obstetrics and Gynecology, University of California, Los Angeles, CA 90095
Christina Charles-Schoeman: Department of Medicine, University of California, Los Angeles, CA 90095
Srinivasa T. Reddy: Department of Medicine, University of California, Los Angeles, CA 90095; Department of Obstetrics and Gynecology, University of California, Los Angeles, CA 90095; Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095 and
Alan M. Fogelman: Department of Medicine, University of California, Los Angeles, CA 90095
Mohamad Navab: Department of Medicine, University of California, Los Angeles, CA 90095
Otto O. Yang: Department of Medicine, University of California, Los Angeles, CA 90095; Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA 90095

Journal volume & issue: Vol. 52, no. 12
pp. 2341 – 2351

Abstract

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Most current assays of HDL functional properties are cell-based. We have developed a fluorometric biochemical assay based on the oxidation of dihydrorhodamine 123 (DHR) by HDL. This cell-free assay assesses the intrinsic ability of HDL to be oxidized by measuring increasing fluorescence due to DHR oxidation over time. The assay distinguishes the oxidative potential of HDL taken from different persons, and the results are reproducible. Direct comparison of this measurement correlated well with results obtained using a validated cell-based assay (r2= 0.62, P < 0.001). The assay can be scaled from a 96-well format to a 384-well format and, therefore, is suitable for high-throughput implementation. This new fluorometric method offers an inexpensive, accurate, and rapid means for determining the oxidative properties of HDL that is applicable to large-scale clinical studies.

Published in Journal of Lipid Research

ISSN: 0022-2275 (Print); 1539-7262 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Chemistry: Organic chemistry: Biochemistry
Website: https://www.journals.elsevier.com/journal-of-lipid-research

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Abstract

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