Hartati Yeni Wahyuni, Komala Dina Ratna, Hendrati Diana, Gaffar Shabarni, Hardianto Ari, Sofiatin Yulia, Bahti Husein Hernandi
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Bandung, Indonesia.
Department of Public Health, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia.
R Soc Open Sci. 2021 Feb 17;8(2):202040. doi: 10.1098/rsos.202040.
Epithelial sodium channel (ENaC) is a transmembrane protein that has an essential role in maintaining the levels of sodium in blood plasma. A person with a family history of hypertension has a high enough amount of ENaC protein in the kidneys or other organs, so that the ENaC protein acts as a marker that a person is susceptible to hypertension. An aptasensor involves aptamers, which are oligonucleotides that function similar to antibodies, as sensing elements. An electrochemical aptasensor for the detection of ENaC was developed using a screen-printed carbon electrode (SPCE) which was modified by electrodeposition of cerium oxide (CeO). The aptamer immobilization was via the streptavidin-biotin system. The measurement of changes in current of the active redox [Fe(CN)] was carried out by differential pulse voltammetry. The surfaces of SPCE and SPCE/CeO were characterized using scanning electron microscopy, voltammetry and electrochemical impedance spectroscopy. The Box-Behnken experimental optimization design revealed the streptavidin incubation time, aptamer incubation time and streptavidin concentrations were 30 min, 30 min and 10.8 µg ml, respectively. Various concentrations of ENaC were used to obtain the linearity range of 0.05-3.0 ng ml, and the limits of detection and quantification were 0.012 ng ml and 0.038 ng ml, respectively. This aptasensor method has the potential to measure the ENaC protein levels in urine samples as well as to be a point-of-care device.
上皮钠通道(ENaC)是一种跨膜蛋白,在维持血浆钠水平方面起着至关重要的作用。有高血压家族史的人在肾脏或其他器官中具有足够高的ENaC蛋白量,因此ENaC蛋白可作为一个人易患高血压的标志物。适配体传感器涉及适配体,适配体是功能类似于抗体的寡核苷酸,作为传感元件。使用通过氧化铈(CeO)电沉积修饰的丝网印刷碳电极(SPCE)开发了一种用于检测ENaC的电化学适配体传感器。适配体通过链霉亲和素-生物素系统固定。通过差分脉冲伏安法测量活性氧化还原[Fe(CN)]电流的变化。使用扫描电子显微镜、伏安法和电化学阻抗谱对SPCE和SPCE/CeO的表面进行了表征。Box-Behnken实验优化设计表明,链霉亲和素孵育时间、适配体孵育时间和链霉亲和素浓度分别为30分钟、30分钟和10.8μg/ml。使用不同浓度的ENaC获得线性范围为0.05 - 3.0 ng/ml,检测限和定量限分别为0.012 ng/ml和0.038 ng/ml。这种适配体传感器方法有潜力测量尿液样本中的ENaC蛋白水平,并且有望成为一种即时检测设备。