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(e ATRP)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/nano Au)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-mercaptophenyl2-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e ATRP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. 0×102mg/L,检测限为6. 8×10-8mg/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e ATRP和含金属蛋白的敏感检测均有重要意义。
Sensitive detection of metal-containing proteins is extremely important in pathology.Superoxidedismutase(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 forelectrochemically mediated atom transfer radical polymerization(eATRP)reaction to prepare SOD PIPs.Thismethod 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 goldelectrode(Au/L-cys/nanoAu)as one working electrode for catalytic reduction of SOD,and initiator(4-mereaptophenyl 2-bromo-2-methylpropanoate,4-HTP-Br)modified Au electrode as another workingelectrode 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 abiosensor for the determination of SOD by differential pulse voltammetry(DPV)measurement.The linearrange is 1.0 x10-7 to 100 mg/L with the detection limit of 6.8 x10-8 mg/L(S/N=3),the correlationa 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 proteinsand the sensitive detection of metal-containing proteins.
作者
刘雨桐
赵梦元
李思雨
杨艺菲
孙越
LIU Yutong;ZHAO Mengyuan;LI Siyu;YANG Yifei;SUN Yue(School of Chemistry and Chemical Engineering,Liaoning Normal University,Dalian,Liaoning 116029,China)
出处
《应用化学》
CAS
CSCD
北大核心
2019年第5期585-594,共10页
Chinese Journal of Applied Chemistry
基金
国家自然科学基金(21304041)
辽宁省教育厅重点实验室(L201683656)
大连市高层次人才创新支持项目(2016RQ047)资助~~
