论文部分内容阅读
设计了一种基于金属离子螯合法固定蛋白质的新方法.首先电化学聚合苯胺(ANI)/邻氨基苯甲酸(OAA)得到在中性溶液中具有导电性的聚(苯胺-邻氨基苯甲酸)(PAOAA)共聚物膜,并对膜进行了SEM、EDS表征.随后膜负载Cu2+,Cu2+作为螯合离子通过配位作用固定过氧化氢酶(Cat).用EDS测量了铜的负载量为0.49%.脉冲伏安法(DPV)研究了Cat在金电极表面的固定过程,显示随着ANI/OAA在电极表面聚合成膜、Cat在修饰电极表面固定,电极在10mmol/LK3Fe(CN)6/K4Fe(CN)6(Fe(CN)63-/4-)溶液中的扫描峰电流由裸电极的121μA依次降到92.6、71.8μA,表明电极表面修饰的PAOAA膜和固定的Cat阻碍了电荷的传递.电化学阻抗法(EIS)对固定过程中电极表面阻抗变化的研究也得到同样结论.用线性扫描伏安法(LSV)研究了固定到电极表面后Cat对H2O2的催化还原活性.最后以计时电流法(CA)作为检测手段,Cat修饰电极作为探针实现了对H2O2的定量检测,同时该电极显示出良好的重建性.
A new method of immobilizing protein based on metal ion chelation was designed. First, aniline (ANI) / anthranilic acid (OAA) was electrochemically polymerized to obtain poly (aniline-anthranilic acid) (PAOAA) copolymer membranes were synthesized and characterized by SEM and EDS.The membranes were then loaded with Cu2 + and Cu2 + as chelating ions to fix catalase (Cat) through coordination.The copper loading was 0.49 %. The immobilization of Cat on the gold electrode surface was investigated by pulsed voltammetry (DPV). Cats were fixed on the surface of the modified electrode as the ANI / OAA was polymerized on the surface of the electrode. The electrode was immersed in 10mmol / L K3Fe (CN) The peak current in K4Fe (CN) 6 (Fe (CN) 63- / 4-) solution decreased from 121μA at the bare electrode to 92.6, 71.8μA in turn, indicating that electrode surface-modified PAOAA membrane and immobilized Cat blocked the charge Electrochemical impedance spectroscopy (EIS) was also applied to study the change of surface impedance of electrode during immobilization.Catalytic reduction activity of Cat on H2O2 was studied by linear sweep voltammetry (LSV) Chronoamperometry (CA) as a detection means, Cat modified electrode as a probe to achieve the right H2O2 quantitative detection, while the electrode showed good reconstruction.