Erdem Arzum, Senturk Huseyin, Yildiz Esma, Maral Meltem
Analytical Chemistry Department, Faculty of Pharmacy, Ege University, Bornova, 35100 Izmir, Türkiye.
Analytical Chemistry Department, Faculty of Pharmacy, Ege University, Bornova, 35100 Izmir, Türkiye.
Int J Biol Macromol. 2024 Nov;281(Pt 1):136233. doi: 10.1016/j.ijbiomac.2024.136233. Epub 2024 Oct 1.
COVID-19 is an infectious disease caused by the SARS-CoV-2 virus, which rapidly spread worldwide and resulted in a pandemic. Efficient and sensitive detection techniques have been devised since the onset of the epidemic and continue to be improved at present. Due to the crucial role of the SARS-CoV-2 S1 protein in facilitating the virus's entry into cells, efforts in detection and treatment have primarily centered upon this protein. In this study, a rapid, ultrasensitive, disposable, easy-to-use, cost-effective next generation biosensor based on optimized aptamer (Optimer, OPT) was developed by using a disposable pencil graphite electrode (PGE) and applied for the impedimetric determination of SARS-CoV-2 S1 protein. The S1 protein interacted with the OPT in the solution phase and then immobilized onto the PGE surface. Subsequently, measurements using electrochemical impedance spectroscopy (EIS) were conducted in a solution containing a redox probe of 1 mM [Fe(CN)]. Under optimum conditions, the limit of detection (LOD) for the S1 protein in buffer medium at concentrations ranging from 10 to 10 ag/mL was calculated as 8.80 ag/mL (0.11 aM). The selectivity of the developed biosensor was studied against MERS-CoV-S1 protein (MERS) and Influenza Hemagglutinin antigen (HA). Furthermore, the application of the biosensor in artificial saliva medium is demonstrated. The LOD was also calculated in artificial saliva medium in the concentration range of 10-10 ag/mL and calculated as 2.01 ag/mL (0.025 aM). This medium was also used to assess the selectivity of optimized-aptamer based biosensor.
新型冠状病毒肺炎（COVID-19）是由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的一种传染病，该病毒在全球迅速传播并导致了大流行。自疫情爆发以来，人们已经设计出了高效且灵敏的检测技术，并且目前仍在不断改进。由于SARS-CoV-2 S1蛋白在促进病毒进入细胞方面的关键作用，检测和治疗工作主要围绕该蛋白展开。在本研究中，通过使用一次性铅笔石墨电极（PGE），开发了一种基于优化适配体（Optimer，OPT）的快速、超灵敏、一次性使用、易于操作且经济高效的下一代生物传感器，并将其应用于SARS-CoV-2 S1蛋白的阻抗测定。S1蛋白在溶液相中与OPT相互作用，然后固定在PGE表面。随后，在含有1 mM [Fe(CN)]氧化还原探针的溶液中使用电化学阻抗谱（EIS）进行测量。在最佳条件下，缓冲介质中浓度范围为10至10 ag/mL的S1蛋白的检测限（LOD）计算为8.80 ag/mL（0.11 aM）。研究了所开发生物传感器对中东呼吸综合征冠状病毒S1蛋白（MERS）和流感血凝素抗原（HA）的选择性。此外，还展示了该生物传感器在人工唾液介质中的应用。在人工唾液介质中浓度范围为10 - 10 ag/mL时也计算了LOD，结果为2.01 ag/mL（0.025 aM）。该介质还用于评估基于优化适配体的生物传感器的选择性。
