超氧化物歧化酶催化-电化学调控的原子转移自由基聚合方法制备分子印迹聚合物

doi:10.11944/j.issn.1000-0518.2019.05.180258

摘要/Abstract

摘要：

病理学中对含金属蛋白质的敏感检测极其重要。本文以超氧化物歧化酶(SOD)作为金属蛋白,SOD既作为模板分子又作为催化剂进行电化学调控的原子转移自由基聚合(eATRP)反应制备蛋白质印迹聚合物(PIPs),用于SOD电化学生物传感器。该方法不需要过渡金属离子,具有制备简单、节约试剂、保护环境等优点。我们选用L-半胱氨酸和纳米金修饰的金电极(Au/L-cys/nanoAu)作为工作电极将氧化型SOD催化还原为还原型SOD,利用还原型SOD的Cu(Ⅰ)粒子,在引发剂4-硫苯基-2-溴-2-甲基丙酸酯(4-mercaptophenyl 2-bromo-2-methylpropanoate,4-HTP-Br)修饰的金电极上调控丙烯酰胺、N,N-亚甲基双丙烯酰胺的eATRP聚合制备SOD PIPs。利用循环伏安法(CV)和X射线光电子能谱(XPS)方法对其进行了表征。通过微分脉冲伏安法(DPV),在最优的条件下利用此修饰电极对溶液中的SOD进行检测,线性响应范围为1.0×10^-7~1.0×10² mg/L,检测限为6.8×10^-8 mg/L(S/N=3),相关系数为0.995。与其它检测SOD的方法相比,该方法具有更宽的线性范围和较低的检测限。本研究对于制备PIPs,用蛋白质催化的eATRP和含金属蛋白的敏感检测均有重要意义。

关键词: 蛋白质印迹聚合物, 电化学调控原子转移自由基聚合, 超氧化物歧化酶, 传感器

Abstract:

Sensitive detection of metal-containing proteins is extremely important in pathology. Superoxide dismutase(SOD) was represented as metal-containing protein to prepare protein imprinted polymers(PIPs) and used for electrochemical biosensor. SOD was used as a template molecule and as a catalyst for electrochemically mediated atom transfer radical polymerization(eATRP) reaction to prepare SOD PIPs. This method does not require transition metal ions and owns the advantages of simple preparation, reagent saving, environmental protection, etc. In this experiment, we selected L-cysteine(L-cys) and nano gold modified gold electrode(Au/L-cys/nanoAu) as one working electrode for catalytic reduction of SOD, and initiator (4-mercaptophenyl 2-bromo-2-methylpropanoate, 4-HTP-Br) modified Au electrode as another working electrode for SOD PIPs modification. The electrode modified with PIPs was characterized by cyclic voltammetry(CV) and X ray photoelectron spectroscopy(XPS). Finally, the PIPs-modified electrode was used as a biosensor for the determination of SOD by differential pulse voltammetry(DPV) measurement. The linear range is 1.0×10^-7 to 100 mg/L with the detection limit of 6.8×10^-8 mg/L(S/N=3), the correlation coefficient is 0.995. Compared with other methods for detecting SOD, this method has a wider linear range and lower detection limits. This work is significant for the preparation of PIPs, eATRP catalyzed by proteins and the sensitive detection of metal-containing proteins.

Key words: protein imprinted polymer, electrochemically-mediated atom transfer radical polymerization, superoxide dismutase, sensor

刘雨桐,赵梦元,李思雨,杨艺菲,孙越. 超氧化物歧化酶催化-电化学调控的原子转移自由基聚合方法制备分子印迹聚合物[J]. 应用化学, 2019, 36(5): 585-594.

LIU Yutong,ZHAO Mengyuan,LI Siyu,YANG Yifei,SUN Yue. Preparation of Molecularly Imprinted Polymers by Superoxide Dismutase-Catalyzed Electrochemically-Mediated Atom Transfer Radical Polymerization[J]. Chinese Journal of Applied Chemistry, 2019, 36(5): 585-594.

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