One-pot assembling pyrroloquinoline quinone glucose dehydrogenase with polydopamine to overcome the reproducibility issues of layer-by-layer electrode development†
{"title":"One-pot assembling pyrroloquinoline quinone glucose dehydrogenase with polydopamine to overcome the reproducibility issues of layer-by-layer electrode development†","authors":"Alessandra Cimino, Shixin Wang, Verdiana Marchianò, Angelo Tricase, Angela Stefanachi, Eleonora Macchia, Blanca Cassano, Luisa Torsi, Xiaoming Zhang and Paolo Bollella","doi":"10.1039/D5SD00053J","DOIUrl":null,"url":null,"abstract":"<p >The reproducibility of enzyme-based biosensors remains a critical challenge, particularly in clinical and wearable applications. Here, we present a novel one-pot polydopamine (PDA)-assisted immobilization strategy for pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ-GDH) on graphite electrodes to address the limitations of conventional layer-by-layer (LbL) methods. The (PQQ-GDH/PDA)<small><sub>OPA</sub></small>/G platform demonstrated a uniform and nanostructured enzyme–polymer matrix, confirmed by SEM and spectroscopic characterization, resulting in enhanced surface coverage and enzyme stabilization. Electrochemical analyses revealed an onset potential of +0.19 ± 0.01 V and a maximum current of 0.87 ± 0.08 μA in the presence of glucose. Amperometric calibration yielded a linear range of 0.4–1.2 mM, a sensitivity of 0.47 μA mM<small><sup>−1</sup></small>, and a low detection limit of 26 ± 2 μM. Michaelis–Menten kinetic analysis provided an <em>I</em><small><sub>max</sub></small> of 1.13 ± 0.07 μA and a <em>K</em><small><sup>app</sup></small><small><sub>M</sub></small> of 3.11 ± 0.59 mM. Reproducibility was excellent, with relative standard deviations below 8% for all key parameters. The biosensor retained full functionality under physiological conditions (pH 7.2, 37 °C) and exhibited high selectivity against common interferents, including dopamine, uric acid, and ascorbic acid, with signal variations below 5%. Remarkably, the sensor maintained stable responses in artificial serum for over 67 days, confirming its long-term operational stability. These findings highlight the one-pot PDA-based approach as a scalable, reproducible, and biocompatible platform for next-generation glucose biosensors suitable for real-world biomedical monitoring.</p>","PeriodicalId":74786,"journal":{"name":"Sensors & diagnostics","volume":" 9","pages":" 750-758"},"PeriodicalIF":4.1000,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12231961/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sensors & diagnostics","FirstCategoryId":"1085","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2025/sd/d5sd00053j","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
Abstract
The reproducibility of enzyme-based biosensors remains a critical challenge, particularly in clinical and wearable applications. Here, we present a novel one-pot polydopamine (PDA)-assisted immobilization strategy for pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ-GDH) on graphite electrodes to address the limitations of conventional layer-by-layer (LbL) methods. The (PQQ-GDH/PDA)OPA/G platform demonstrated a uniform and nanostructured enzyme–polymer matrix, confirmed by SEM and spectroscopic characterization, resulting in enhanced surface coverage and enzyme stabilization. Electrochemical analyses revealed an onset potential of +0.19 ± 0.01 V and a maximum current of 0.87 ± 0.08 μA in the presence of glucose. Amperometric calibration yielded a linear range of 0.4–1.2 mM, a sensitivity of 0.47 μA mM−1, and a low detection limit of 26 ± 2 μM. Michaelis–Menten kinetic analysis provided an Imax of 1.13 ± 0.07 μA and a KappM of 3.11 ± 0.59 mM. Reproducibility was excellent, with relative standard deviations below 8% for all key parameters. The biosensor retained full functionality under physiological conditions (pH 7.2, 37 °C) and exhibited high selectivity against common interferents, including dopamine, uric acid, and ascorbic acid, with signal variations below 5%. Remarkably, the sensor maintained stable responses in artificial serum for over 67 days, confirming its long-term operational stability. These findings highlight the one-pot PDA-based approach as a scalable, reproducible, and biocompatible platform for next-generation glucose biosensors suitable for real-world biomedical monitoring.