Materials and Engineering Research Institute, Sheffield Hallam University, Sheffield S1 1WB, UK.
Department of Chemistry, The University of Sheffield, Sheffield S3 7HF, UK.
Int J Mol Sci. 2021 Nov 24;22(23):12701. doi: 10.3390/ijms222312701.
This work is a continuation of our research into the development of simple, reliable, and cost-effective methods for the early diagnosis of prostate cancer (PCa). The proposed method is based on the electrochemical detection of the PCA3 biomarker of PCa (long non-coded RNA transcript expressed in urine) using a specific aptamer labeled with a redox group (methylene blue). The electrochemical measurements (cyclic voltammograms) obtained from electrodes functionalized with the aptamer were complemented in this work by another biosensing technique: total internal reflection ellipsometry (TIRE). In addition to proving the concept of the detection of PCA3 in low concentrations down to 90 pM, this study improved our understanding of the processes by which PCA3 binds to its specific aptamer. The high specificity of the binding of PCA3 to the aptamer was assessed by studying the binding kinetics, which yielded an affinity constant (K) of 2.58 × 10 M. Additional XPS measurements confirmed the strong covalent binding of aptamers to gold and showed spectral features associated with PCA3 to aptamer binding.
这项工作是我们研究如何开发简单、可靠且经济有效的前列腺癌 (PCa) 早期诊断方法的延续。所提出的方法基于使用带有氧化还原基团（亚甲蓝）标记的特定适体来电化学检测 PCa 的 PCA3 生物标志物（在尿液中表达的长非编码 RNA 转录物）。在这项工作中，通过另一种生物传感技术：全内反射椭圆测量法（TIRE）补充了通过适体功能化的电极获得的电化学测量（循环伏安法）。除了证明在低至 90 pM 的低浓度下检测 PCA3 的概念之外，这项研究还增进了我们对 PCA3 与其特定适体结合的过程的理解。通过研究结合动力学来评估 PCA3 与适体结合的高特异性，得出亲和力常数（K）为 2.58×10 M。额外的 XPS 测量证实了适体与金的强共价结合，并显示了与 PCA3 与适体结合相关的光谱特征。
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