Scientific article
English

Development of immobilized enzyme reactors based on human recombinant cytochrome P450 enzymes for phase I drug metabolism studies

Published inJournal of chromatography, vol. 1206, no. 1, p. 2-10
Publication date2008
Abstract

Two cytochrome P450 (CYP)-based immobilized enzyme reactors (IMERs) were developed to perform automated on-line phase I drug metabolism studies. For this purpose, biotinylated recombinant CYP2D6 or CYP3A4 reconstituted systems were anchored to the surface of two monolithic mini-columns (2 mm x 6 mm I.D.), which had been covalently grafted with NeutrAvidin. After optimization of immobilization conditions, the obtained IMERs were integrated on-line into a LC hyphenated to an electrospray ionization MS/MS system. Studies with probe substrates and a known competitive inhibitor were performed, showing the potential of CYP-based IMERs in drug metabolism. In the optimized conditions, ca. 15 experiments were carried out with each bioreactor.

Keywords
  • Avidin
  • Bioreactors
  • Biotinylation
  • Chromatography, Liquid/instrumentation/methods
  • Cytochrome P-450 CYP2D6/antagonists & inhibitors/metabolism
  • Cytochrome P-450 CYP3A/antagonists & inhibitors/metabolism
  • Drug Evaluation, Preclinical/methods
  • Enzyme Inhibitors/pharmacology
  • Enzyme Stability
  • Enzymes, Immobilized/metabolism
  • Humans
  • Kinetics
  • Microsomes, Liver/chemistry
  • Recombinant Proteins/metabolism
  • Spectrometry, Mass, Electrospray Ionization/instrumentation/methods
Citation (ISO format)
NICOLI, Raul et al. Development of immobilized enzyme reactors based on human recombinant cytochrome P450 enzymes for phase I drug metabolism studies. In: Journal of chromatography, 2008, vol. 1206, n° 1, p. 2–10. doi: 10.1016/j.chroma.2008.05.080
Main files (1)
Article (Accepted version)
accessLevelRestricted
Identifiers
ISSN of the journal0021-9673
592views
5downloads

Technical informations

Creation03/26/2009 9:32:00 AM
First validation03/26/2009 9:32:00 AM
Update time03/14/2023 3:03:41 PM
Status update03/14/2023 3:03:41 PM
Last indexation10/29/2024 11:23:53 AM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack