title: Evaluation of immobilized hFMO3 on magnetic nanoparticles by capillary zone electrophoresis.
abstract: OBJECTIVE
  In this paper, we developed a method to immobilize human flavin-containing monooxygenase-3 (hFMO3) using glutaraldehyde as a cross-linker onto amino-functionalized magnetic nanoparticles.
  
  
  METHODS
  All the analyses were done using capillary electrophoresis coupled with a diode array detector using clozapine as a substrate.
  
  
  RESULTS
  The apparent Kmwith clozapine as substrate and inhibition of hFMO3 by methimazole were explored for immobilized hFMO3 and were found to be comparable to literature values. The immobilized enzyme could be used three-times continuously at 37°C with no loss in enzyme activity.
  
  
  CONCLUSIONS
  A method to immobilize hFMO3 on magnetic nanoparticles has been described and evaluated in terms of enzyme activity, inhibition, pH stability and reusability.
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
authors.forename: Pranov
authors.initials: P
authors.lastname: Ramana
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
authors.forename: Ward
authors.initials: W
authors.lastname: Herman
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Hiroux
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
authors.forename: Erwin
authors.initials: E
authors.lastname: Adams
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, Drug delivery & Disposition, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 921, 3000 Leuven, Belgium.
authors.forename: Patrick
authors.initials: P
authors.lastname: Augustijns
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
authors.forename: Ann
authors.initials: A
authors.lastname: Van Schepdael
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D010105:Oxygenases / Q000378:metabolism*; D013696:Temperature
chemical_list: D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)
keywords: NADPH; amino-functionalized magnetic nanoparticles; capillary electrophoresis; clozapine; glutaraldehyde conjugation; hFMO3; immobilization; magnetic nanoparticles; methimazole; sodium borohydride