In the current study, an electrochemical biosensing signal amplification system was utilized with thionine-chitosan-gold nanoparticles (Chit-GNPs) that absorbed horseradish peroxidase (HRP) and anti-His tagged protein monoclonal antibody derived from Balb/c mice. In addition, transmission electron microscopy (TEM) was used to characterize the nanogold solution and atomic force microscopy (AFM) was used to characterize the sensor assembly. To evaluate the quality of the immunosensor, the amperometric I-t curve method was applied to determine His-IL23 in PBS. The results indicated that the response current exhibited an optimal linear correlation with the His-IL23 concentration that ranged from 0.01 to 10³ ng/ml. The lowest detection limit was noted at 3.3 pg/ml (S/N = 3). The linear equation was deduced as follows: △I = 0.02lgC + 0.037 (R² = 0.9628). Moreover, it was validated with high sensitivity, reproducibility and rapid response. Apparently, the immunosensor may be a very useful tool for the detection and quantification of His-tagged proteins. In addition, the signal amplification system can be used for the preparation of other immunosensors and to assist in bioassays.

中文翻译：

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基于硫氨酸-壳聚糖/金纳米颗粒/辣根过氧化物酶的检测组氨酸标记蛋白的新方法

在当前的研究中，电化学生物传感信号放大系统与吸收辣根过氧化物酶（HRP）和衍生自Balb / c小鼠的抗His标记蛋白单克隆抗体的硫氨酸-壳聚糖-金纳米颗粒（Chit-GNP）一起使用。此外，透射电子显微镜（TEM）用于表征纳米金溶液，原子力显微镜（AFM）用于表征传感器组件。为了评估免疫传感器的质量，应用了安培型It曲线法确定了PBS中的His-IL23。结果表明，响应电流与His-IL23浓度呈最佳线性相关，范围为0.01至10 ³  ng / ml。最低检测限为3.3 pg / ml（S / N = 3）。线性方程式推导如下：△I = 0.02lgC + 0.037（R ²  = 0.9628）。此外，它还具有高灵敏度，可重复性和快速响应的特点。显然，免疫传感器可能是检测和定量His标记蛋白的非常有用的工具。此外，信号放大系统可用于制备其他免疫传感器并协助进行生物测定。