自阵列液滴适配体电化学传感器检测细胞因子干扰素-γ  

Self-patterned Droplet Aptasensor for Electrochemical Detection of Human Interferon-gamma

作　　者：王莹菲 孟潇 鞠熀先 刘颖 WANG Ying-Fei;MENG Xiao;JU Huang-Xian;LIU Ying(State Key Laboratory of Analytical Chemistry for Life Science,School of Chemistry and Chemical Engineering,Nanjing University,Nanjing 210023,China;Chemistry and Biomedicine Innovation Center,Nanjing University,Nanjing 210023,China)

机构地区：[1]南京大学化学化工学院,生命分析化学国家重点实验室,南京210023 [2]南京大学化学和生物医药创新研究院,南京210023

出　　处：《分析化学》2022年第11期1743-1749,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.22022405,21974064)资助。

摘　　要：设计了一种基于核酸适配体检测人干扰素-γ(hIFN-γ)的液滴自阵列化电化学生物传感器。通过光刻技术在镀金玻片上加工形成三电极体系阵列,并利用光点击反应对三电极体系阵列进行亲疏水图案化修饰,形成亲水性的加样检测区和疏水性周边以实现检测样品的液滴自阵列化。电活性物质亚甲基蓝(MB)标记的发卡型核酸适配体通过金硫键自组装于工作电极表面,初始状态时,MB和电极表面距离较近,显示电化学还原峰。核酸适配体对hIFN-γ有特异性结合能力,与hIFN-γ结合后,核酸适配体空间构象发生变化,使MB分子远离电极表面,引起其电化学信号显著减小。采用方波伏安法(SWV)进行检测,在hIFN-γ浓度10.0~125.0 ng/mL范围内,此传感器具有良好的线性响应,检出限(3σ)为7.5 ng/mL。此自阵列液滴适配体传感器可用于干扰素hIFN-γ的快速灵敏检测,对研究免疫细胞的受刺激响应与实现细胞因子释放相关疾病的即时检测具有重要意义。An electrochemical aptasensor for analysis of human interferon-gamma(hIFN-γ) based on self-patterned droplet on substrate that is particularly suitable for point-of-care testing is reported. After fabricating the gold electrode system on glass slide via photolithography, the glass slide is functionalized with alkyne-silane. Sequential thiol-yne photoclick reactions are subsequently performed to develop hydrophilic 1-thioglycerol microwells around the electrodes with the surrounding regions modified with hydrophobic 1 H,1 H,2 H,2 H-perfluorodecanethiol. The hydrophilic microwells around electrode facilitate sample loading and enrichment by concentrating large-volume sample solution into small self-patterned droplet. The working electrode is further functionalized with thiolated aptamer for hIFN-γ. The hairpin structured aptamer molecule contains redox reporter molecule methylene blue, and the binding of analyte to aptamer-modified electrode inhibits electron transfer from redox reporters to the electrode and causes electrochemical redox signal decrease. The aptasensor demonstrates a linear relationship for hIFN-γ proteins from 10.0 ng/mL to 125.0 ng/mL with detection limit(3σ) of 7.5 ng/mL. The self-patterned droplet aptasensor developed here provides a convenient method for rapid cytokine hIFN-γ detection with high efficiency and accuracy, and demonstrates great potential for the rapid diagnosis of related diseases in a point-of-care testing manner.

关 键 词：细胞因子 适配体传感器 适配体 亲水性 疏水性 

分 类 号：O657.1[理学—分析化学] R446.61[理学—化学]