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Cell-free Biochemical Fluorometric Enzymatic Assay for High-throughput Measurement of Lipid Peroxidation in High Density Lipoprotein
07:29

Cell-free Biochemical Fluorometric Enzymatic Assay for High-throughput Measurement of Lipid Peroxidation in High Density Lipoprotein

Published on: October 12, 2017

Nitrated apolipoprotein A-I, a potential new cardiovascular marker, is markedly increased in low high-density

Ahmed Bakillah1

  • 1GlaxoSmithKline, Research and Development, Clinical Pharmacology and Discovery Medicine, King of Prussia, PA, USA. abakillah@hotmail.com

Clinical Chemistry and Laboratory Medicine
|December 6, 2008
PubMed

Insights

A new assay detects elevated nitrated apolipoprotein A-I in patients with cardiovascular disease. This method may help identify individuals at higher risk for atherosclerosis by monitoring nitrated apolipoprotein A-I levels.

Area of Science:

  • Biochemistry
  • Cardiovascular Medicine
  • Biomarker Discovery

Background:

  • Oxidative stress is implicated in coronary artery disease (CAD) pathogenesis.
  • Elevated nitrated apolipoprotein A-I (nit-apoA-I) is found in high-density lipoproteins (HDL) from CAD patients.
  • Quantifying plasma nit-apoA-I could correlate with atherosclerosis risk.

Purpose of the Study:

  • To develop and validate a method for quantifying plasma nitrated apolipoprotein A-I.
  • To assess the relationship between nit-apoA-I levels and cardiovascular disease status.
  • To explore nit-apoA-I as a potential biomarker for inflammatory vascular disease.

Main Methods:

  • Western blot analysis was used to detect nit-apoA-I.
  • A novel sandwich enzyme-linked immunosorbent assay (ELISA) was developed for quantification.
  • Assay precision was validated by comparing ELISA and Western blot results (r²=0.96).

Main Results:

  • Significant 3-fold elevations in plasma nit-apoA-I were observed in low HDL cholesterol subjects compared to high HDL cholesterol subjects.
  • Both plasma and serum samples are suitable for nit-apoA-I quantification via ELISA.
  • Higher nit-apoA-I content in HDL particles correlated negatively with circulating apolipoprotein A-I levels.

Conclusions:

  • A rapid, sensitive, and reproducible ELISA method for measuring nit-apoA-I has been established.
  • This novel method requires no sample manipulation, offering practical advantages.
  • The assay is proposed as a valuable tool for monitoring circulating nit-apoA-I and assessing its potential as a biomarker for inflammatory vascular diseases like atherosclerosis.
Abstract

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