Abstract
In this chapter, we describe the development and fabrication of gold microelectrodes based on silicon by silicon technology, for multiplex detection of cytokines. Cytokines have become a crucial biomarker for the identification of end-stage heart failure (ESHF) for patients during early phase of left ventricular assisted device (LVAD) implantation. The microelectrode device consists of three gold working microelectrodes that were activated and 4-aminophenylacetic acid (CMA) was electroaddressed onto individual gold WEs. The carboxylic acid functionalities of the diazotated aromatic amine were activated through carbodiimide chemistry and anti-interleukin-10 monoclonal antibodies (anti-IL-10 mAb) were immobilized onto the transducers surface. The interaction between the antibody-antigen (Ab-Ag) was characterized by electrochemical impedance spectroscopy (EIS). Here, Nyquist plots have shown a stepwise variation due to the charge transfer resistance (Rct) between the Ab activated surfaces with the detection of the human IL-10. For early expression monitoring, commercial proteins of human IL-10 were analyzed between 1 pg/mL and 100 pg/mL. The protein concentrations within the linear range of 1–50 pg/mL were detected and these values formulated a sensitivity of 0.008 (pg/mL)\(^{-1}\) (\(R^2 = 0.9840\)). These preliminary results demonstrated that the developed biosensor was sensitive to the detection of human IL-10 and the calculated limit of detection (LOD) was measured at 0.156 (pg/mL)\(^{-1}\). To validate the biosensors response, the experiment was repeated several times on different gold working WEs by applying the same conditions. The overall relative standard deviation percentage (% R.S.D.) was 4.9% which demonstrates the successful fabrication for the detection of human IL-10 through diazonium salt electroreduction.
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Acknowledgements
We acknowledge the funding through the European Union’s Horizon 2020 and innovation programme entitled (An integrated POC solution for non-invasive diagnosis and therapy monitoring of Heart Failure patients, KardiaTool) under grant agreement No. 768686. This work partially supported by POC4Allergies # ERA-PerMed-2019 and PHC PROCOPE #40544QH projects.
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Lee, M. et al. (2021). Impedimetric Detection of Human Interleukin 10 on Diazonium Salt Electroaddressed Gold Microelectrode Surfaces. In: Kanoun, O., Derbel, N. (eds) Advanced Sensors for Biomedical Applications. Smart Sensors, Measurement and Instrumentation, vol 38. Springer, Cham. https://doi.org/10.1007/978-3-030-71225-9_7
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