Ramana Pranov, Schejbal Jan, Houthoofd Kristof, Martens Johan, Adams Erwin, Augustijns Patrick, Glatz Zdenĕk, Van Schepdael Ann
Department of Pharmaceutical and Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Leuven, Belgium.
Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
Electrophoresis. 2018 Apr;39(7):981-988. doi: 10.1002/elps.201700434. Epub 2018 Jan 19.
In this paper, we demonstrate the effectiveness of a new 3D printed magnet holder that enables capture of magnetic microparticles in commercially available capillary electrophoresis equipment with a liquid or air based coolant system. The design as well as the method to capture magnetic microparticles inside the capillary are discussed. This setup was tested at temperature and pH values suitable for performing enzymatic reactions. To demonstrate its applicability in CE- immobilized microenzyme reactors (IMER) development, human flavin-containing monooxygenase 3 and bovine serum albumin were immobilized on amino functionalized magnetic microparticles using glutaraldehyde. These microparticles were subsequently used to perform in-line capillary electrophoresis with clozapine as a model substrate. This setup could be used further to establish CE-IMERs of other drug metabolic enzymes in a commercially available liquid based capillary coolant system. The CE-IMER setup was successful, although a subsequent decrease in enzyme activity was observed on repeated runs.
在本文中，我们展示了一种新型3D打印磁体固定器的有效性，该固定器能够在配备基于液体或空气冷却系统的市售毛细管电泳设备中捕获磁性微粒。讨论了毛细管内部捕获磁性微粒的设计及方法。此装置在适合进行酶促反应的温度和pH值条件下进行了测试。为证明其在毛细管电泳固定化微酶反应器（IMER）开发中的适用性，使用戊二醛将人含黄素单加氧酶3和牛血清白蛋白固定在氨基功能化磁性微粒上。随后，以氯氮平作为模型底物，使用这些微粒进行在线毛细管电泳。该装置可进一步用于在市售的基于液体的毛细管冷却系统中建立其他药物代谢酶的毛细管电泳 - IMER。毛细管电泳 - IMER装置是成功的，不过在重复运行时观察到酶活性随后有所下降。
