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Updated: Jun 28, 2026

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A high-throughput screen for endothelial lipase using HDL as substrate.

Paul M Keller¹, Timothy Rust, Dennis J Murphy

¹GSK Screening & Compound Profiling, Collegeville, Pennsylvania, USA.

Journal of Biomolecular Screening

|June 21, 2008

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Endothelial lipase (EL) inhibition may increase good cholesterol (HDL-C) levels, potentially reducing cardiovascular disease risk. Researchers developed a high-throughput assay to discover novel EL inhibitors for therapeutic development.

Area of Science:

Biochemistry
Cardiovascular Biology
Drug Discovery

Background:

Endothelial lipase (EL) is a lipase enzyme uniquely expressed in endothelial cells, preferring phospholipids as substrates.
EL plays a role in regulating high-density lipoprotein cholesterol (HDL-C) levels, with animal models showing an inverse correlation.
Elevated HDL-C is associated with reduced cardiovascular disease risk due to its role in reverse cholesterol transport.

Purpose of the Study:

To develop a high-throughput screening (HTS) assay for identifying inhibitors of endothelial lipase.
To discover novel compounds that modulate EL activity for potential therapeutic applications in cardiovascular disease.

Main Methods:

A coupled fluorescent assay was developed utilizing HDL as the native substrate for EL.

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High-Density Lipoprotein-Specific Phospholipid Efflux Assay

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Published on: September 30, 2025

The assay measures free fatty acids released by EL hydrolysis, coupled to a resorufin-based fluorescent detection system.

A 1536-well, 5-microliter format assay was employed for high-throughput screening against the GSK compound collection.

Main Results:

A functional high-throughput assay for endothelial lipase was successfully established and validated.
A large-scale HTS campaign was conducted, identifying initial screening hits.
Novel mechanism-of-action studies were developed and applied to characterize a screening hit.

Conclusions:

The developed HTS assay provides a robust platform for discovering endothelial lipase inhibitors.
Inhibition of EL is a promising strategy for increasing HDL-C levels and mitigating cardiovascular disease.
Further investigation of identified hits is warranted to develop EL-targeted therapeutics.