Biomedical Engineering Laboratory, Federal University of Pernambuco, Recife, Brazil.
BioMark/ISEP, School of Engineering, Polytechnic Institute of Porto, Porto, Portugal.
Biosens Bioelectron. 2016 Mar 15;77:978-85. doi: 10.1016/j.bios.2015.10.068. Epub 2015 Oct 28.
A nano-molecularly imprinted polymer (N-MIP) assembled on a screen-printed electrode for the cardiac troponin T (cTnT) was developed. The biomimetic surface was obtained by a co-polymer matrix assembled on the reduced graphene oxide (RGO) electrode surface. The cTnT active sites were engineered using pyrrole and carboxylated pyrrole that was one-step electropolymerized jointly with cTnT by cyclic voltammetry. The stepwise preparation of the biomimetic surface was characterized by cyclic and differential pulse voltammetries using the ferrocyanide/ferricyanide as redox probe. Structural and morphological characterization was also performed. The optimal relation of pyrrole and pyrrole-3-acid carboxylic to perform the cTnT biomimetic nanosurface was obtained at 1:5 ratio. The analytical performance of cTnT N-MIP performed by differential pulse voltammetry showed a linear range from 0.01 to 0.1 ngmL(-1) (r=0.995, p«0.01), with a very low limit of detection (0.006 ngmL(-1)). The synergic effect of conductive polymer and graphene forming 3D structures of reactive sites resulted in a N-MIP with excellent affinity to cTnT binding (KD=7.3 10(-13) molL(-1)). The N-MIP proposed is based on a simple method of antibody obtaining with a large potential for point-of-care testing applications.
一种基于纳米印迹聚合物 (N-MIP) 的心脏肌钙蛋白 T (cTnT) 传感器被开发出来。该仿生表面是通过共聚物基质在还原氧化石墨烯 (RGO) 电极表面组装得到的。cTnT 的活性位点是通过电聚合吡咯和羧基化吡咯来构建的,这两种物质可以通过循环伏安法与 cTnT 一步电聚合。采用亚铁氰化钾/铁氰化钾作为氧化还原探针,通过循环伏安法和差分脉冲伏安法对仿生表面的逐步制备进行了表征。还进行了结构和形态表征。通过实验得出,在吡咯和吡咯-3-羧酸的比例为 1:5 时,能够实现 cTnT 仿生纳米表面的最佳关系。通过差分脉冲伏安法对 cTnT N-MIP 的分析性能进行测试,结果表明线性范围为 0.01 至 0.1 ngmL(-1)(r=0.995,p«0.01),检测限非常低(0.006 ngmL(-1))。导电聚合物和石墨烯形成的 3D 结构的协同效应导致了对 cTnT 具有优异结合亲和力的 N-MIP(KD=7.3 10(-13) molL(-1))。所提出的 N-MIP 基于获得抗体的简单方法,具有用于即时检测应用的巨大潜力。