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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Autores principales:	Theodoros Kelesidis, Christian K Roberts, Diana Huynh, Otoniel Martínez-Maza, Judith S Currier, Srinivasa T Reddy, Otto O Yang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7a035253c04b45528d912af1954e3995
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spelling	oai:doaj.org-article:7a035253c04b45528d912af1954e39952021-11-25T05:54:45ZA high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.1932-620310.1371/journal.pone.0111716https://doaj.org/article/7a035253c04b45528d912af1954e39952014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0111716https://doaj.org/toc/1932-6203Current 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<0.001) and cell-free assays (r = 0.46, p<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<0.01) and subendocardial viability ratio (r = -0.21, p = 0.05) and physiological parameters such as metabolic and anthropometric parameters (p<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.Theodoros KelesidisChristian K RobertsDiana HuynhOtoniel Martínez-MazaJudith S CurrierSrinivasa T ReddyOtto O YangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 11, p e111716 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Theodoros Kelesidis Christian K Roberts Diana Huynh Otoniel Martínez-Maza Judith S Currier Srinivasa T Reddy Otto O Yang A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
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<0.001) and cell-free assays (r = 0.46, p<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<0.01) and subendocardial viability ratio (r = -0.21, p = 0.05) and physiological parameters such as metabolic and anthropometric parameters (p<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.
format	article
author	Theodoros Kelesidis Christian K Roberts Diana Huynh Otoniel Martínez-Maza Judith S Currier Srinivasa T Reddy Otto O Yang
author_facet	Theodoros Kelesidis Christian K Roberts Diana Huynh Otoniel Martínez-Maza Judith S Currier Srinivasa T Reddy Otto O Yang
author_sort	Theodoros Kelesidis
title	A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
title_short	A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
title_full	A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
title_fullStr	A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
title_full_unstemmed	A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.
title_sort	high throughput biochemical fluorometric method for measuring lipid peroxidation in hdl.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/7a035253c04b45528d912af1954e3995
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A high throughput biochemical fluorometric method for measuring lipid peroxidation in HDL.

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