Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses (IZI-BB), Potsdam-Golm, Germany.
Institute of Biochemistry and Biology, University of Potsdam, Potsdam-Golm, Germany.
Electrophoresis. 2023 Jun;44(11-12):956-967. doi: 10.1002/elps.202300010. Epub 2023 Mar 16.
Dielectrophoresis (DEP) is an AC electrokinetic effect that is proven to be effective for the immobilization of not only cells, but also of macromolecules, for example, antibodies and enzyme molecules. In our previous work, we have already demonstrated the high catalytic activity of immobilized horseradish peroxidase after DEP. To evaluate the suitability of the immobilization method for sensing or research in general, we want to test it for other enzymes, too. In this study, glucose oxidase (GOX) from Aspergillus niger was immobilized on TiN nanoelectrode arrays by DEP. Fluorescence microscopy showed the intrinsic fluorescence of the immobilized enzymes flavin cofactor on the electrodes. The catalytic activity of immobilized GOX was detectable, but a fraction of less than 1.3% of the maximum activity that was expected for a full monolayer of immobilized enzymes on all electrodes was stable for multiple measurement cycles. Therefore, the effect of DEP immobilization on the catalytic activity strongly depends on the used enzyme.
介电泳(DEP)是一种交流电动效应,已被证明不仅可以有效地固定细胞,还可以固定大分子,例如抗体和酶分子。在我们之前的工作中,已经证明了固定化辣根过氧化物酶在 DEP 后的高催化活性。为了评估固定化方法在传感或一般研究中的适用性,我们还希望对其他酶进行测试。在这项研究中,黑曲霉葡萄糖氧化酶(GOX)通过 DEP 固定在 TiN 纳米电极阵列上。荧光显微镜显示了固定在电极上的酶黄素辅因子的固有荧光。可以检测到固定化 GOX 的催化活性,但在多次测量循环中稳定的只有不到预期固定化酶单层最大活性的 1.3%。因此,DEP 固定化对催化活性的影响强烈取决于所用的酶。