Issue 12, 2021

Sensitive electrochemical immunosensor using a bienzymatic system consisting of β-galactosidase and glucose dehydrogenase

Abstract

Bienzymatic systems are often used with electrochemical affinity biosensors to achieve high signal levels and/or low background levels. It is important to select two enzymes whose reactions do not exhibit mutual interference but have similar optimal conditions. Here, we report a sensitive electrochemical immunosensor based on a bienzymatic system consisting of β-galactosidase (Gal, a hydrolase enzyme) and flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH, a redox enzyme). Both enzymes showed high activities at neutral pH, the reactions catalyzed by them did not exhibit mutual interference, and the electrochemical-enzymatic redox cycling based on FAD-GDH coupled with enzymatic amplification by Gal enabled high signal amplification. Among the three amino-hydroxy-naphthalenes and 4-aminophenol (potential Gal products), 4-amino-1-naphthol showed the highest signal amplification. Glucose, as an electro-inactive, stable reducing agent for redox cycling, helped in achieving low background levels. Our bienzymatic system could detect parathyroid hormone at a detection limit of ∼0.2 pg mL−1, implying that it can be used for highly sensitive electrochemical detection of parathyroid hormone and other biomarkers in human serum.

Graphical abstract: Sensitive electrochemical immunosensor using a bienzymatic system consisting of β-galactosidase and glucose dehydrogenase

Supplementary files

Article information

Article type
Paper
Submitted
02 abr 2021
Accepted
06 mai 2021
First published
13 mai 2021

Analyst, 2021,146, 3880-3887

Sensitive electrochemical immunosensor using a bienzymatic system consisting of β-galactosidase and glucose dehydrogenase

S. Park, S. Seo, N. Lee, Y. H. Yoon and H. Yang, Analyst, 2021, 146, 3880 DOI: 10.1039/D1AN00562F

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