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Related Experiment Video

Updated: May 20, 2026

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

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Lipid-sensing high-throughput ApoA-I assays.

Anita Niedziela-Majka1, Latesh Lad, Jeffrey W Chisholm

  • 1Gilead Sciences Inc., 333 Lakeside Dr., Foster City, CA 94404, USA. anita.majka@gilead.com

Journal of Biomolecular Screening
|July 20, 2012
PubMed
Summary

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Developed new nonradioactive assays to measure Apolipoprotein A-I (ApoA-I) lipidation, crucial for cholesterol metabolism and preventing atherosclerosis. These high-throughput methods offer improved sensitivity and dynamic range over traditional assays.

Area of Science:

  • Biochemistry
  • Cardiovascular Biology
  • Assay Development

Background:

  • Apolipoprotein A-I (ApoA-I) is key to high-density lipoprotein (HDL) formation and cholesterol efflux.
  • Defective lipidation of ApoA-I, mediated by ABCA1, is linked to atherosclerosis.
  • Existing radioactive assays for ApoA-I lipidation lack throughput and dynamic range.

Purpose of the Study:

  • To develop sensitive, nonradioactive, high-throughput assays for measuring ApoA-I lipidation.
  • To overcome limitations of current radioactive cholesterol transport assays.

Main Methods:

  • Developed a homogeneous time-resolved fluorescence resonance energy transfer (TR-FRET) assay using labeled ApoA-I.
  • Developed a discontinuous assay utilizing the label-free Epic biosensor platform.

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Last Updated: May 20, 2026

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

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Published on: October 12, 2017

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  • Both assays measure the incorporation of lipids into ApoA-I.

    • Main Results:

    • The TR-FRET assay showed decreased signal proportional to increased lipid-to-ApoA-I ratio, indicating sensitivity to lipidation.
    • The Epic assay demonstrated a resonant wavelength shift proportional to lipid association with ApoA-I.
    • Both assays successfully quantified ApoA-I lipidation in a high-throughput, nonradioactive manner.

    Conclusions:

    • New TR-FRET and Epic assays provide sensitive, high-throughput measurement of ApoA-I lipidation.
    • These assays can advance research into cholesterol metabolism and atherosclerosis.
    • Nonradioactive methods offer advantages over traditional assays for studying HDL formation.