Xiangya School of Pharmaceutical Sciences, Central South University, No. 172, Tongzipo Road, Changsha 410013, Hunan Province, China.
Changsha Cinotohi Technology Co., Ltd, No. 229, West Tongzipo Road, Changsha, Hunan 410013, China.
Anal Methods. 2021 Oct 8;13(38):4442-4451. doi: 10.1039/d1ay01105g.
As an essential biochemical indicator in the fields of pregnancy and oncology, human chorionic gonadotropin (HCG) can be evaluated using colloidal gold immunochromatographic paper and quantified using a biochemical analyzer based on the principle of the antibody sandwich method. In view of the inaccuracy of the former and the complication of the latter, this study constructed an accurate, sensitive and simple unlabeled biosensor based on peptide aptamer CGGGPPLRINRHILTR for HCG detection. Molecular Operating Environment (MOE) was used to simulate the aptamer and protein docking, and western blot (WB) was used to verify the binding effect and ratio. The peptide aptamer was characterized and was then reduced with tris-(2-carboxyethyl)-phosphine hydrochloride (TCEP). After electrochemical deposition of chloroauric acid on the screen-printed electrode (SPE), the aptamer was self-assembled on the electrode surface under optimal conditions. The active site of the electrode surface was blocked with 6-mercapto-1-hexanol (MCH) and BSA. The electrochemical impedance spectrum (EIS) was used to quantify HCG in the matrix. Showing a good linear relationship in the range of 5-1500 mIU mL, with a detection limit of 1 mIU mL, the biosensor remained stable at room temperature for 14 days. Because of its small size, stability, sensitivity and accuracy, this biosensor has great potential to become a portable diagnostic device for HCG.
作为妊娠和肿瘤领域的重要生化指标,人绒毛膜促性腺激素(HCG)可以使用胶体金免疫层析试纸进行评估,并基于抗体夹心法的原理使用生化分析仪进行定量。鉴于前者的不准确性和后者的复杂性,本研究构建了一种基于肽适体 CGGGPPLRINRHILTR 的准确、敏感、简单的无标记生物传感器,用于检测 HCG。分子操作环境(MOE)用于模拟适体和蛋白质对接,Western blot(WB)用于验证结合效果和比例。对肽适体进行表征,然后用三(2-羧乙基)膦盐酸盐(TCEP)还原。在丝网印刷电极(SPE)上电沉积氯金酸后,在最佳条件下,适体在电极表面自组装。用 6-巯基-1-己醇(MCH)和 BSA 封闭电极表面的活性位点。电化学阻抗谱(EIS)用于定量基质中的 HCG。在 5-1500 mIU mL 的范围内呈现良好的线性关系,检测限为 1 mIU mL,该生物传感器在室温下稳定 14 天。由于其体积小、稳定性好、灵敏度高、准确性高,该生物传感器具有成为 HCG 便携式诊断设备的巨大潜力。