[en] An original method called SIEFED (specific immunological extraction followed by enzymatic detection) was developed for the specific detection of the activity of equine myeloperoxidase (MPO). The method consists of the extraction of MPO from aqueous solutions by immobilized anti-MPO antibodies followed by washing (to eliminate proteins and interfering molecules) and measurement of MPO activity using a detection system containing a fluorogenic substrate, hydrogen peroxide, and nitrite as reaction enhancer. The SIEFED technique was applied to study active MPO in horse biological fluids and the effects of 2 polyphenolic molecules, curcumin and resveratrol, on MPO activity. The detection limit of the SIEFED was 0.23 mU/ml. The SIEFED exhibited good precision with intra-assay and interassay coefficients of variation below 10% and 20%, respectively, for MPO activities ranging from 0.25 to 6.4 mU/ml. The activity of MPO was generally higher than 1 mU/ml in the fluids collected from horses with inflammatory diseases. Curcumin and resveratrol exerted a dose-dependent inhibition on MPO activity and, as they were removed before the enzymatic detection of MPO, the results suggest a direct drug-nzyme interaction or an enzyme structure modification by the drug. The SIEFED is a new tool that would be useful for specific detection of active MPO in complex media and for selection of MPO activity modulators.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Kohnen, Stephan ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Deby-Dupont, Ginette; Université de Liège - ULiège > Centre de l'Oxygène : Recherche et Développement (C.O.R.D.)
Grulke, Sigrid ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés
Deby, Carol; Centre de Biochimie de l'Oxygène
Serteyn, Didier ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Language :
English
Title :
A specific method for measurement of equine active myeloperoxidase in biological samples and in in vitro tests
Publication date :
July 2006
Journal title :
Journal of Veterinary Diagnostic Investigation
ISSN :
1040-6387
Publisher :
Amer Assoc Veterinary Laboratory Diagnosticians Inc, Turlock, United States - California
Volume :
18
Issue :
4
Pages :
326-334
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
MPOEQUI
Funders :
DGTRE - Région wallonne. Direction générale des Technologies, de la Recherche et de l'Énergie
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