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This paper presents a study on the impedance spectroscopy methods applied to 2-electrode and 3-electrode biosensors. While 3-electrode electrochemical biosensors have been extensively studied and applied in various applications, the 2-electrode configuration of electrochemical biosensors still lacks comprehensive investigation for real-world applications. In particular, the miniaturization of the 3-electrode configuration for integration into microfluidic devices has been explored due to the relatively limited number of previous studies. A commercial screen-printed gold electrode was employed to develop immunosensors for bovine serum albumin (BSA) detection. The comparisons were performed on non-Faradaic and Faradic immunosensors corresponding to 2-electrode and 3-electrode configurations through impedance measurements. The results show the performance of both types of sensors in detecting BSA protein. While the 2-electrode configuration-based sensor can detect the BSA protein with a sensitivity of 12 Ω/µM, the 3-electrode configuration-based sensor shows superior results with a sensitivity of 7.2 kΩ/µM. Besides, the results also indicate that the 2-electrode configuration-based sensor can easily be miniaturized to integrate into a microfluidic channel to develop lab-on-a-chip systems. Although the 3-electrode configuration demonstrates advancements compared to the 2-electrode configuration, the 2-electrode system based on non-Faradaic processes still shows high potential for future applications. [ABSTRACT FROM AUTHOR]

AbstractThis study presents a screen-printed carbon electrode surface functionalization to immobilize proteins. The process involves the deposition of gold nanoparticles on the carbon electrode and subsequent surface modification to immobilize bovine serum albumin–fluorescein isothiocyanate conjugate (BSA-FITC). A combination of conducting polymer and gold nanoparticles was employed to enhance the electrode contact area and current transmission. The electrode surface morphology and properties were evaluated by scanning electron microscopy (SEM) and Raman spectroscopy, showing the porous polyaniline (PANI) film has been formed on the carbon electrode surface with the network of dendritic nanofibers. Gold nanoparticles were evenly distributed on the PANI film to form a nanocomposite material layer on the carbon electrode. The observed size of these gold nanoparticles was significantly smaller than those on the electrode without a PANI film. Additionally, the current transmission of this composite material layer has been verified by cyclic voltammetry (CV). The results show that the amplitude of the redox peak was doubled compared to the bare carbon electrode and gold nanoparticle-only deposited electrode. Moreover, the success of the process was verified by fluorescence and cyclic voltammetry. The fluorescent green color observed under the microscope indicated that BSA-FITC was on the electrode surface. The change of CV signal after each step verified the formation of a new layer on the electrode surface. These results are anticipated to play an important role in developing high-sensitivity immunosensors in the future. [ABSTRACT FROM AUTHOR]