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Cholesterol Efflux Assay
Published on: March 6, 2012
A simple method for the determination of the cholesterol esterase activity
Agnieszka Ewa Stępień1, Mykhailo Gonchar
1Department of Biotechnology, Institute of Applied Biotechnology and Basic Science; and Centre of Applied Biotechnology and Basic Sciences, University of Rzeszow, Kolbuszowa, Poland.
Acta Biochimica Polonica
|September 21, 2013
Summary
This study introduces a precise, low-cost assay for determining cholesterol esterase (CEH) activity using human serum. The method is valuable for quality control of cholesterol enzymatic kits and screening bacterial enzyme producers.
Area of Science:
- Biochemistry
- Enzymology
- Analytical Chemistry
Background:
- Cholesterol esterase (CEH) plays a crucial role in cholesterol metabolism.
- Accurate determination of CEH activity is essential for diagnostic and research applications.
- Existing methods may be costly or lack precision.
Purpose of the Study:
- To develop a novel, cost-effective, and precise spectrophotometric assay for quantifying cholesterol esterase (CEH) activity.
- To utilize human serum as a natural substrate source for CEH in the assay.
- To validate the assay for quality control of enzyme preparations and microbial screening.
Main Methods:
- The assay adapts Allain's spectrophotometric method for cholesterol determination.
- It employs cholesterol oxidase and peroxidase enzymes.
- A key modification involves using 3,5-dichloro-dihydroxybenzenesulfonic acid (DHBS) as the phenolic chromogen and human serum as the substrate source.
Main Results:
- The proposed method accurately determines CEH activity.
- It offers a low-cost alternative to existing assays.
- The assay demonstrates high precision, suitable for quantitative analysis.
Conclusions:
- The developed assay provides a reliable and economical tool for measuring CEH activity.
- It is applicable for quality control of cholesterol esterase preparations used in diagnostic kits.
- The method facilitates the screening of bacterial strains for novel enzyme production.
