Cholesterol Assay Based on Recombinant Cholesterol Oxidase, ABTS, and Horseradish Peroxidase

Alexey V Sviridov; Mikhail V Karpov; Victoria V Fokina; Marina V Donova

doi:10.1007/978-1-0716-3385-4_9

Cholesterol Assay Based on Recombinant Cholesterol Oxidase, ABTS, and Horseradish Peroxidase

Methods Mol Biol. 2023:2704:157-171. doi: 10.1007/978-1-0716-3385-4_9.

Authors

Alexey V Sviridov¹, Mikhail V Karpov¹, Victoria V Fokina¹, Marina V Donova¹

Affiliation

¹ G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Federal Research Center "Pushchino Center for Biological Research of the Russian Academy of Sciences", Pushchino, Russia.

PMID: 37642843
DOI: 10.1007/978-1-0716-3385-4_9

Abstract

Cholesterol determination by cholesterol oxidase reaction is a fast, convenient, and highly specific approach with widespread use in clinical diagnostics. Routinely, endpoint measurements with 4-aminophenazone or 4-aminoantipyrine as chromogens and sodium cholate, surfactants, or alcohols as solubilizing agents are used. Here we describe a novel kinetic method to determine cholesterol in 0.05-0.75 mM range in neutral or acidic buffers by use of recombinant cholesterol oxidase from Nocardioides simplex in a coupled reaction with horseradish peroxidase, ABTS as a chromogen, and methyl-β-cyclodextrin as a solubilizing agent.

Keywords: ABTS; Cholesterol; Cholesterol oxidase; Enzymatic measurement; Horseradish peroxidase; Methyl-β-cyclodextrin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cholesterol Oxidase*
Cholesterol*
Horseradish Peroxidase

Substances

Horseradish Peroxidase
2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid
Cholesterol Oxidase
azo rubin S
Cholesterol