A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL

Current cell-based assays for determining the functional properties of high-density lipoproteins (HDL) have limitations. We report here the development of a new, robust fluorometric cell-free biochemical assay that measures HDL lipid peroxidation (HDLox) based on the oxidation of the fluorochrome Am...

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Published in:PloS one 2014-11, Vol.9 (11), p.e111716-e111716
Main Authors: Kelesidis, Theodoros, Roberts, Christian K, Huynh, Diana, Martínez-Maza, Otoniel, Currier, Judith S, Reddy, Srinivasa T, Yang, Otto O
Format: Article
Language:English
Subjects:
Adolescent
Adult
Animal models
Animals
Anthropometry
Apolipoproteins
Arteriosclerosis
Assaying
Atherosclerosis
Biology and Life Sciences
Cardiovascular diseases
Carotid Intima-Media Thickness
Cholesterol
Coronary Artery Disease - blood
Coronary Artery Disease - metabolism
Correlation analysis
Female
Fluorometry - methods
Gynecology
Heart diseases
High density lipoprotein
High-Throughput Screening Assays - methods
HIV
HIV Infections - blood
HIV Infections - metabolism
HIV-1 - isolation & purification
Human immunodeficiency virus
Humans
Immunology
Laboratories
Lipid Peroxidation
Lipids
Lipoproteins
Lipoproteins (high density)
Lipoproteins, HDL - blood
Lipoproteins, HDL - chemistry
Lipoproteins, HDL - metabolism
Low density lipoprotein
Male
Medical research
Medicine
Medicine and Health Sciences
Metabolism
Mice, Inbred C57BL
Middle Aged
Obstetrics
Oxazines - analysis
Oxazines - metabolism
Oxidation
Peroxidation
Studies
Viability
Viruses
Young Adult
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Reddy, Srinivasa T
Yang, Otto O
description Current cell-based assays for determining the functional properties of high-density lipoproteins (HDL) have limitations. We report here the development of a new, robust fluorometric cell-free biochemical assay that measures HDL lipid peroxidation (HDLox) based on the oxidation of the fluorochrome Amplex Red. HDLox correlated with previously validated cell-based (r = 0.47, p
doi_str_mv 10.1371/journal.pone.0111716
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Other authors have no conflicts of interest to declare. This manuscript is related to the Provision Patent Application UCLA Case UCLAP123P/2014-425-1 entitled “High Throughput Biochemical Fluorometric Method for Measuring HDL Redox Activity”. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.</notes><notes>Conceived and designed the experiments: TK STR OY. Performed the experiments: TK DH CKR. Analyzed the data: TK CKR. Contributed reagents/materials/analysis tools: TK CKR JSC OMM STR OY. Wrote the paper: TK STR OY.</notes><abstract>Current cell-based assays for determining the functional properties of high-density lipoproteins (HDL) have limitations. We report here the development of a new, robust fluorometric cell-free biochemical assay that measures HDL lipid peroxidation (HDLox) based on the oxidation of the fluorochrome Amplex Red. HDLox correlated with previously validated cell-based (r = 0.47, p&lt;0.001) and cell-free assays (r = 0.46, p&lt;0.001). HDLox distinguished dysfunctional HDL in established animal models of atherosclerosis and Human Immunodeficiency Virus (HIV) patients. Using an immunoaffinity method for capturing HDL, we demonstrate the utility of this novel assay for measuring HDLox in a high throughput format. Furthermore, HDLox correlated significantly with measures of cardiovascular diseases including carotid intima media thickness (r = 0.35, p&lt;0.01) and subendocardial viability ratio (r = -0.21, p = 0.05) and physiological parameters such as metabolic and anthropometric parameters (p&lt;0.05). In conclusion, we report the development of a new fluorometric method that offers a reproducible and rapid means for determining HDL function/quality that is suitable for high throughput implementation.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25368900</pmid><doi>10.1371/journal.pone.0111716</doi><oa>free_for_read</oa></addata></record>
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identifier ISSN: 1932-6203
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1932-6203
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source Publicly Available Content Database; PubMed Central
subjects Adolescent
Adult
Animal models
Animals
Anthropometry
Apolipoproteins
Arteriosclerosis
Assaying
Atherosclerosis
Biology and Life Sciences
Cardiovascular diseases
Carotid Intima-Media Thickness
Cholesterol
Coronary Artery Disease - blood
Coronary Artery Disease - metabolism
Correlation analysis
Female
Fluorometry - methods
Gynecology
Heart diseases
High density lipoprotein
High-Throughput Screening Assays - methods
HIV
HIV Infections - blood
HIV Infections - metabolism
HIV-1 - isolation & purification
Human immunodeficiency virus
Humans
Immunology
Laboratories
Lipid Peroxidation
Lipids
Lipoproteins
Lipoproteins (high density)
Lipoproteins, HDL - blood
Lipoproteins, HDL - chemistry
Lipoproteins, HDL - metabolism
Low density lipoprotein
Male
Medical research
Medicine
Medicine and Health Sciences
Metabolism
Mice, Inbred C57BL
Middle Aged
Obstetrics
Oxazines - analysis
Oxazines - metabolism
Oxidation
Peroxidation
Studies
Viability
Viruses
Young Adult
title A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL
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