Koenig Meike, König Ulla, Eichhorn Klaus-Jochen, Müller Martin, Stamm Manfred, Uhlmann Petra
Department of Nanostructured Materials, Leibniz Institute of Polymer Research Dresden, Dresden, Germany.
Department of Analytics, Leibniz Institute of Polymer Research Dresden, Dresden, Germany.
Front Chem. 2019 Mar 7;7:101. doi: 10.3389/fchem.2019.00101. eCollection 2019.
Herein, we report on the use of a combined setup of quartz-crystal microbalance, with dissipation monitoring and spectroscopic ellipsometry, to comprehensively investigate the covalent immobilization of an enzyme to a polymer layer. All steps of the covalent reaction of the model enzyme glucose oxidase with the poly(acrylic acid) brush by carbodiimide chemistry, were monitored . Data were analyzed using optical and viscoelastic modeling. A nearly complete collapse of the polymer chains was found upon activation of the carboxylic acid groups with N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide and N-Hydroxysuccinimide. The reaction with the amine groups of the enzyme occurs simultaneously with re-hydration of the polymer layer. Significantly more enzyme was immobilized on the surface compared to physical adsorption at similar conditions, at the same pH. It was found that the pH responsive swelling behavior was almost not affected by the presence of the enzyme.
在此，我们报道了使用一种结合了石英晶体微天平、耗散监测和椭圆偏振光谱的装置，来全面研究酶与聚合物层的共价固定化。通过碳二亚胺化学方法，对模型酶葡萄糖氧化酶与聚丙烯酸刷的共价反应的所有步骤进行了监测。使用光学和粘弹性模型对数据进行了分析。在用N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺和N-羟基琥珀酰亚胺活化羧酸基团后，发现聚合物链几乎完全塌陷。与酶的胺基的反应与聚合物层的再水合同时发生。在相同pH值的相似条件下，与物理吸附相比，表面固定的酶明显更多。发现pH响应性溶胀行为几乎不受酶存在的影响。
