Optimization of Aptamer-Based Electrochemical Biosensor for ATP Detection Using Screen-Printed Carbon Electrode/Gold Nanoparticles (SPCE/AuNP)

Abstract

Electrochemical biosensors are used to detect adenosine triphosphate (ATP) levels, which are involved in a variety of biological processes, such as regulating cellular metabolism and biochemical pathways. Therefore, this research aims to develop an aptamer-based electrochemical biosensor with Screen Printed Carbon Electrode/gold nanoparticles (SPCE/AuNP) and collect data as well as information related to ATP detection. The modification of SPCE with AuNP increased the analyte’s binding sensitivity and biocompatibility. The aptamer was selected based on its excellent bioreceptor characteristics. Furthermore, aptamer–SH (F1) and aptamer-NH2 (F2) were immobilized on the SPCE/AuNP surface, which had been characterized using SEM, EIS, and DPV. Also, the ATP-binding aptamers were electrochemically characterized using the K3[Fe(CN)6] redox system and Differential Pulse Voltammetry (DPV). According to the optimization results using the Box-Behnken experimental design, the ideal conditions obtained from the factors influencing the experiment were the F1 concentration and incubation time of 4 µM and 24 h, respectively, as well as F1/F2/ATP incubation time of 7.5 min. Meanwhile, for the range of 0.1 to 100 µM, the detection (LoD) and quantification (LoQ) limits were 7.43 and 24.78 µM, respectively. Therefore, this aptasensor method can be used to measure ATP levels in real samples.

Advances in electrochemical biosensors employing carbon-based electrodes for detection of biomarkers in diabetes mellitus

“…The biosensor was fabricated by electrodeposition of gold nanoparticles (AuNPs) on SPCE. AuNPs serve to immobilize biomolecules and enhance the current response due to gold's high conductivity and electrocatalytic properties [ 152 ]. The thiolated aptamer interacts with AuNPs, forming a covalent bond.…”

“…Recently, Mulyani et al . [ 152 ] developed an electrochemical biosensor that selectively detects ATP using an aptamer from Kashefi-Kheyrabadi's research [ 157 ]. The thiolated aptamer was immobilized on SPCE through bonding between thiol groups and gold groups deposited by the drop-casting method.…”

Detection of Biomarker Using Aptasensors to Determine the Type of Diabetes

“…Recent research from Mulyani in 2022 also supports the research of Kashefi-Khernyabadi (2013); Mulyani conducted electrochemical sensor research using the ATP-binding aptamer with the differential pulse voltammogram (DPV) method. The voltammogram characterization results show that the peak current in ATP is lower than in UTP, CTP, and GTP [108]. Rustaman et al (2023) conducted research on the specificity of aptasensors for diabetes mellitus detection using in silico methods.…”

Transfer RNA Mutation Associated with Type 2 Diabetes Mellitus

“…Recently, a tool for detecting ATP levels has been developed using an aptamer-based chemical biosensor with screen-printed carbon electrode/gold nano-particles (SPCE/AuNP). After undergoing a series of tests, this aptasensor method has shown potential for sample analysis [ 130 ].…”