Article (Scientific journals)
Fully automated electrophoretically mediated microanalysis for CYP1A1 activity monitoring optimized by multivariate approach
Farcas, Elena; Servais, Anne-Catherine; Lamalle, Caroline et al.
2016In Electrophoresis, 37 (2), p. 248-255
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Keywords :
EMMA; CYP1A1 activity monitoring; ethoxycoumarin; metabolisation; inhibition
Abstract :
[en] In this study, a fully automatized in-capillary system was developed to monitor the activity of CYP1A1 in physiological conditions. Ethoxycoumarin, the selected substrate, undergoes an in-line bioreaction in the presence of CYP1A1 supersomes and NADPH as co-factor, giving rise to hydroxycoumarin, the product that was assayed. The optimization of the experimental conditions was supported by the application of a design of experiment, providing a better understanding of electrophoretic mixing parameters that influence the metabolic reactions. The results obtained in optimal conditions were compared not only to those achieved after off-line metabolization but also with liver microsomes. Finally, inhibition studies were conducted showing an important decrease of hydroxycoumarin formation using apigenin as CYP1A1 potent inhibitor. This study demonstrates the usefulness of our in-line system for the fully automated in vitro metabolism studies and the screening of new CYP1A1 inhibitors.
Research center :
CIRM - Centre Interdisciplinaire de Recherche sur le Médicament - ULiège
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Farcas, Elena ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Servais, Anne-Catherine  ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Lamalle, Caroline ;  Université de Liège > Département de pharmacie > Analyse des médicaments
CHIAP, Patrice ;  Centre Hospitalier Universitaire de Liège - CHU > Pharmacologie clinique
Pochet, Lionel
Fillet, Marianne ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Fully automated electrophoretically mediated microanalysis for CYP1A1 activity monitoring optimized by multivariate approach
Publication date :
2016
Journal title :
Electrophoresis
ISSN :
0173-0835
eISSN :
1522-2683
Publisher :
Vch Publishers, Weinheim, Germany
Volume :
37
Issue :
2
Pages :
248-255
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 07 October 2015

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