Ramana Pranov, Herman Ward, Hiroux Charlotte, Adams Erwin, Augustijns Patrick, Van Schepdael Ann
Department of Pharmaceutical & Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
Department of Pharmaceutical & Pharmacological Sciences, Drug delivery & Disposition, KU Leuven - University of Leuven, Herestraat 49, O&N2, PB 921, 3000 Leuven, Belgium.
Bioanalysis. 2017 Feb;9(3):289-296. doi: 10.4155/bio-2016-0151. Epub 2017 Jan 13.
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.
The apparent K 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.
A method to immobilize hFMO3 on magnetic nanoparticles has been described and evaluated in terms of enzyme activity, inhibition, pH stability and reusability.
在本文中,我们开发了一种方法,以戊二醛作为交联剂,将人含黄素单加氧酶-3(hFMO3)固定在氨基功能化磁性纳米颗粒上。
所有分析均采用毛细管电泳结合二极管阵列检测器,以氯氮平作为底物进行。
研究了以氯氮平为底物时固定化hFMO3的表观K值以及甲巯咪唑对hFMO3的抑制作用,发现其与文献值相当。固定化酶在37°C下可连续使用三次,酶活性无损失。
已描述了一种将hFMO3固定在磁性纳米颗粒上的方法,并从酶活性、抑制作用、pH稳定性和可重复使用性方面进行了评估。