【作者】 王海燕； 王明慧； 殷焕顺； 艾仕云；

【Author】 Haiyan Wang;Minghui Wang;Huanshun Yin;Shiyun Ai;College of Chemistry and Material Science, Shandong Agricultural University;

【机构】 山东农业大学化学与材料科学学院；

【摘要】 光电化学生物分析是基于光电化学过程与化学和生物识别过程相结合建立起来的一种新的分析方法。本文研究了一种利用类石墨相碳化氮（g-C₃N₄）、CdS量子点异质结作为光电转换材料,CuO作为指示剂以及抗坏血酸作为电子供体的灵敏的、可用于RNA甲基化的光电化学分析方法。实验中利用抗体与抗原的免疫反应,生物素与亲和素的特定反应以及甲基化RNA与m⁶A抗体的特异性反应成功构建了生物免疫传感器并极大提高了反应的特异性。该生物免疫传感器对甲基化RNA的检测呈现高灵敏度,检出限为3.53 pM,线性方程为I_pc （μA）=-0.13logc （nM）+0.23。并利用该传感器分别对健康人、乳腺癌患者以及治疗后乳腺癌患者体内的RNA甲基化水平进行了研究并对比,进一步扩宽了制备的传感器的应用性。因此,本文所提出的光电化学方法在RNA甲基化的检测方面具有极大地潜力和实用价值。更多还原

【Abstract】 N⁶-methyladenosine（m⁶A） is an enigmatic and abundant internal modification in eukaryotic messenger RNA（m RNA）, which could affect various aspects of RNA metabolism and m RNA translation.Herein, a novel photoelectrochemical（PEC） immunosensor was constructed for m⁶A detection based on the inhibition of Cu²⁺to the photoactivity of CdS QDs/g-C₃N₄ heterojunction, where g-C₃N₄/CdS QDs heterojunction was used as photoactive material, anti-m⁶A antibody as recognition unit for RNA containing m⁶A, Phos-tag-biotin as link unit and avidin functionalized CuO as PEC signal inhibitor.When CuO was captured on electrode through biotin-avidin affinity reaction and treated with HCl, Cu²⁺could be released and CuxS would be formed based on the selective interaction between CdS QDs and Cu²⁺, which could cause an obviously decrease of the photocurrent.Under the optimal detection conditions, the PEC biosensor displayed a linear range of 0.01-10 nM and a low detection limit of 3.53 pM for methylated RNA determination.Furthermore, the developed method can also be used to detect the expression level of m⁶A methylated RNA in serum samples of breast cancer patient before and after operative treatment.The proposed assay strategy has a great potential for detecting the expression methylation level of RNA in real sample.更多还原