The optimization of electrochemical immunosensors to detect epithelial sodium channel as a biomarker of hypertension

Tias Febriana Hanifa Lestari; Riyanto Setiyono; Nina Tristina; Yulia Sofiatin; Yeni Wahyuni Hartati

doi:10.5599/admet.1629

Authors

Tias Febriana Hanifa Lestari Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia https://orcid.org/0000-0002-3486-9615
Riyanto Setiyono Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia
Nina Tristina Department of Clinical Pathology, Faculty of Medicine, Universitas Padjadjaran, Indonesia
Yulia Sofiatin Department of Public Health, Faculty of Medicine, Universitas Padjadjaran, Indonesia
Yeni Wahyuni Hartati Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia https://orcid.org/0000-0003-1463-6352

DOI:

https://doi.org/10.5599/admet.1629

Keywords:

Box-Behnken design, screen-printed carbon electrode, immunosensor, hypertension

Abstract

The epithelial sodium channel (ENaC) is a transmembrane protein that regulates the balance of sodium salt levels in the body through its expression in various tissues. The increase in sodium salt in the body is related to the expression of ENaC, thereby increasing blood pressure. Therefore, overexpression of the ENaC protein can be used as a biomarker for hypertension. The detection of ENaC protein using anti-ENaC in the biosensor system has been optimized with the Box-Behnken experimental design. The steps carried out in this research are screen-printed carbon electrode modification with gold nanoparticles, then anti-ENaC was immobilized using cysteamine and glutaraldehyde. Optimum conditions of the experiment, such as anti-ENaC concentration, glutaraldehyde incubation time, and anti-ENaC incubation time, were optimized using the Box-Behnken experimental design to determine the factors that influence the increase in immunosensor current response and the optimum conditions obtained were then applied to variations in ENaC protein concentrations. The optimum experimental conditions for anti-ENaC concentration were 2.5 µg/mL, the glutaraldehyde incubation time was 30 minutes, and the anti-ENaC incubation time was 90 minutes. The developed electrochemical immunosensor has a detection limit of 0.0372 ng/mL and a quantification limit of 0.124 ng/mL for the ENaC protein concentration range of 0.09375 to 1.0 ng/mL. Thus, the immunosensor generated from this study can be used to measure the concentration of normal urine samples and those of patients with hypertension.

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