Demirbakan Burçak, Kemal Sezgintürk Mustafa
Çanakkale Onsekiz Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, Turkey.
Anal Chim Acta. 2021 Feb 1;1144:43-52. doi: 10.1016/j.aca.2020.12.001. Epub 2020 Dec 4.
In present study, we developed a highly sensitive, electrochemical immunosensor based on fullerene C-modified disposable graphite paper (GP) electrode for determination of Suppression of Tumorigenicity 2 (ST2) in human serum. The synthesis of the ST2 immunosensor was monitored with electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) techniques and single frequency impedance (SFI) technique which is utilized for the specific interaction between anti-ST2 and ST2 antigen. Moreover, the morphological alteration of each GP surface was examined by scanning electron microscopy (SEM), SEM-energy dispersive X-ray spectroscopy (EDX) and atomic force microscopy (AFM). All parameters such as fullerene C concentration, antibody concentration and antibody incubation time were optimized. Analytical characteristics such as linear determination range, repeatability, reproducibility, regeneration and surface coverage were determined for the immunosensor. The ST2 electrochemical immunosensor had excellent repeatability, reproducibility and a wide detection range (from 0.1 fg mL to 100 fg mL). The proposed immunosensor also had low limit of detection (LOD) and limit of quantification (LOQ) values of 0.124 fg mL and 0.414 fg mL, respectively. The proposed immunosensor was applied to real samples to test applicability in clinical practice.
在本研究中,我们基于富勒烯C修饰的一次性石墨纸(GP)电极开发了一种高灵敏度的电化学免疫传感器,用于测定人血清中的抑瘤素2(ST2)。采用电化学阻抗谱(EIS)、循环伏安法(CV)技术和单频阻抗(SFI)技术监测ST2免疫传感器的合成过程,其中SFI技术用于检测抗ST2与ST2抗原之间的特异性相互作用。此外,通过扫描电子显微镜(SEM)、扫描电子显微镜-能量色散X射线光谱(EDX)和原子力显微镜(AFM)对每个GP表面的形态变化进行了检测。对富勒烯C浓度、抗体浓度和抗体孵育时间等所有参数进行了优化。测定了免疫传感器的线性测定范围、重复性、再现性、再生性和表面覆盖率等分析特性。ST2电化学免疫传感器具有出色的重复性、再现性和宽检测范围(从0.1 fg mL至100 fg mL)。所提出的免疫传感器的检测限(LOD)和定量限(LOQ)分别为0.124 fg mL和0.414 fg mL。将所提出的免疫传感器应用于实际样品,以测试其在临床实践中的适用性。