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Evaluation of immobilized hFMO3 on magnetic nanoparticles by capillary zone electrophoresis
Pranov Ramana1, Ward Herman1, Charlotte Hiroux1
1Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
Aim:
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.
Materials & Methods:
All the analyses were done using capillary electrophoresis coupled with a diode array detector using clozapine as a substrate.
Results:
The apparent Km with 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.
Conclusion:
A method to immobilize hFMO3 on magnetic nanoparticles has been described and evaluated in terms of enzyme activity, inhibition, pH stability and reusability.
