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A self-contained 48-well fatty acid oxidation assay
Xiaojun Wang1, Rose Wang, Thomas A Nemcek
1Metabolic Disease Research, Abbott Laboratories, Abbott Park, IL 60064-3502, USA.
Assay and Drug Development Technologies
|April 20, 2004
Summary
This study introduces a novel 48-well assay for measuring beta-oxidation, significantly increasing throughput for drug discovery. The new system effectively captures CO2, aiding research into obesity and insulin resistance treatments.
Area of Science:
- Biochemistry
- Metabolic Research
- Drug Discovery
Background:
- Fatty acid metabolism modulation, particularly beta-oxidation, is a key therapeutic target for obesity and insulin resistance.
- Current beta-oxidation assays have low throughput, hindering drug discovery efforts in this field.
Purpose of the Study:
- To develop and validate a high-throughput 48-well beta-oxidation assay.
- To facilitate the evaluation of compounds targeting fatty acid metabolism.
Main Methods:
- A novel instrument design with connecting chambers for CO2 capture in a sealed 48-well format.
- Measurement of CO2 production from labeled substrates to quantify beta-oxidation.
- Assessment of cellular viability and response to known modulators (e.g., AMP kinase activator).
Main Results:
- The assay demonstrates linear CO2 capture with time and substrate concentration.
- Cellular viability is maintained, and cells respond appropriately to modulators.
- The assay is adaptable to different lipid substrates and cell lines.
Conclusions:
- The developed 48-well beta-oxidation assay offers significantly increased throughput compared to existing methods.
- This assay is a valuable tool for accelerating the discovery of drugs targeting fatty acid metabolism for obesity and insulin resistance.