|Reference : Specific Immuno Extraction Followed by Enzymatic Detection (SIEFED), a new tool to measu...|
|Scientific congresses and symposiums : Paper published in a book|
|Human health sciences : Pharmacy, pharmacology & toxicology|
Human health sciences : Laboratory medicine & medical technology
Life sciences : Veterinary medicine & animal health
Life sciences : Biochemistry, biophysics & molecular biology
|Specific Immuno Extraction Followed by Enzymatic Detection (SIEFED), a new tool to measure active myeloperoxidase in complex media and to screen potential inhibitors of this enzyme.|
|Serteyn, Didier [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux >]|
|Mouithys-Mickalad, Ange [Université de Liège - ULg > > Centre de l'oxygène : Recherche et développement (C.O.R.D.) >]|
|Deby-Dupont, Ginette [Université de Liège - ULg > > Centre de l'Oxygène, Recherche et Développement (CORD) > >]|
|Franck, Thierry [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux >]|
|The 7th International Human Peroxidase Meeting, 22-25 May 2011, Brussels|
|The 7th International Human Peroxidase Meeting|
|22-25 May, 2011|
|[en] SIEFED ; Myeloperoxidase activity ; Pharmacological tool ; enzyme-immunocapture|
|[en] A method called SIEFED (“Specific Immuno Extraction Followed by Enzymatic Detection”) was developed for the specific measurement of myeloperoxidase (MPO) activity without interference of the sample medium. In this method, MPO is first extracted out of aqueous or biological samples by its capture by immobilized anti-MPO antibodies. A washing step then allows to eliminate the solution or biological fluid with interfering materials (proteins, lipids, reducing or oxidizing molecules, ...) and in a last step the activity of MPO bound to its antibodies is measured by using a sensitive detection system containing a fluorogenic substrate, hydrogen peroxide and nitrite as reaction enhancer.
SIEFED is a method of choice to measure easily, quantitatively, and specifically the active part of MPO present in biological samples or complex media. Results obtained with this technique applied to biological samples emphasize the importance to distinguish the total MPO concentration of a sample obtained by ELISA from the active part of MPO, which is the real witness of the oxidant potential of the enzyme.
SIEFED is also a powerful tool to study compounds or natural extracts that could have an inhibitory effect on the activity of MPO. Since the potential inhibitor is first incubated with the enzyme solution, and further eliminated by washing after the immunocapture of MPO, the tested compound or extract cannot interfere with the chromogenic substrate used to measure the activity of MPO or with products derived from the enzyme activity. Thus, an inhibitory effect could only be attributed to a direct interaction of the tested compound with the enzyme, either on the active site or another key position of the structure.
In conclusion, the SIEFED technique opens new perspectives to study pathologies in which the release of active MPO is relevant and to select interesting compounds or extracts able to modulate the MPO activity.
|Centre de l’Oxygène, Recherche et Développement - CORD|
|Researchers ; Professionals ; Students|
|An international patent (EP 1711817 B1) protects the SIEFED technique|
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