Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection
Glucose (Glu) detection, as a fundamental analytical technique, has applications in medical diagnostics, clinical testing, bioanalysis and environmental monitoring. In this work, a solid-phase electrochemiluminescence (ECL) enzyme sensor was developed by immobilizing the ECL emitter in a stable mann...
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MDPI AG
2025-05-01
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| author | Chunyin Wei Yanyan Zheng Fei Yan Lifang Xu |
| author_facet | Chunyin Wei Yanyan Zheng Fei Yan Lifang Xu |
| author_sort | Chunyin Wei |
| collection | DOAJ |
| description | Glucose (Glu) detection, as a fundamental analytical technique, has applications in medical diagnostics, clinical testing, bioanalysis and environmental monitoring. In this work, a solid-phase electrochemiluminescence (ECL) enzyme sensor was developed by immobilizing the ECL emitter in a stable manner within bipolar silica nanochannel array film (bp-SNA), enabling sensitive glucose detection. The sensor was constructed using an electrochemical-assisted self-assembly (EASA) method with various siloxane precursors to quickly modify the surface of indium tin oxide (ITO) electrodes with a bilayer SNA of different charge properties. The inner layer, including negatively charged SNA (n-SNA), attracted the positively charged ECL emitter tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)<sub>3</sub><sup>2+</sup>) via electrostatic interaction, while the outer layer, including positively charged SNA (p-SNA), repelled it, forming a barrier that efficiently concentrated the Ru(bpy)<sub>3</sub><sup>2+</sup> emitter in a stable manner. After modifying the amine groups on the p-SNA surface with aldehyde groups, glucose oxidase (GOx) was covalently immobilized, forming the enzyme electrode. In the presence of glucose, GOx catalyzed the conversion of glucose to hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), which acted as a quencher for the Ru(bpy)<sub>3</sub><sup>2+</sup>/triethanolamine (TPA) system, reducing the ECL signal and enabling quantitative glucose analysis. The sensor exhibited a wide linear range from 10 μM to 7.0 mM and a limit of detection (LOD) of 1 μM (S/N = 3). Glucose detection in fetal bovine serum was realized. By replacing the enzyme type on the electrode surface, this sensing strategy holds the potential to provide a universal platform for the detection of different metabolites. |
| format | Article |
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| issn | 2079-6374 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
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| series | Biosensors |
| spelling | doaj-art-93bb91ada4f147a3a93bb83e81e6420f2025-08-20T02:33:40ZengMDPI AGBiosensors2079-63742025-05-0115533210.3390/bios15050332Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose DetectionChunyin Wei0Yanyan Zheng1Fei Yan2Lifang Xu3Guangxi Medical University Cancer Hospital, Guangxi Medical University, Nanning 530021, ChinaSchool of Chemistry and Chemical Engineering, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaSchool of Chemistry and Chemical Engineering, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaThe Second Affiliated Hospital of Guangxi Medical University, Nanning 530007, ChinaGlucose (Glu) detection, as a fundamental analytical technique, has applications in medical diagnostics, clinical testing, bioanalysis and environmental monitoring. In this work, a solid-phase electrochemiluminescence (ECL) enzyme sensor was developed by immobilizing the ECL emitter in a stable manner within bipolar silica nanochannel array film (bp-SNA), enabling sensitive glucose detection. The sensor was constructed using an electrochemical-assisted self-assembly (EASA) method with various siloxane precursors to quickly modify the surface of indium tin oxide (ITO) electrodes with a bilayer SNA of different charge properties. The inner layer, including negatively charged SNA (n-SNA), attracted the positively charged ECL emitter tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)<sub>3</sub><sup>2+</sup>) via electrostatic interaction, while the outer layer, including positively charged SNA (p-SNA), repelled it, forming a barrier that efficiently concentrated the Ru(bpy)<sub>3</sub><sup>2+</sup> emitter in a stable manner. After modifying the amine groups on the p-SNA surface with aldehyde groups, glucose oxidase (GOx) was covalently immobilized, forming the enzyme electrode. In the presence of glucose, GOx catalyzed the conversion of glucose to hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), which acted as a quencher for the Ru(bpy)<sub>3</sub><sup>2+</sup>/triethanolamine (TPA) system, reducing the ECL signal and enabling quantitative glucose analysis. The sensor exhibited a wide linear range from 10 μM to 7.0 mM and a limit of detection (LOD) of 1 μM (S/N = 3). Glucose detection in fetal bovine serum was realized. By replacing the enzyme type on the electrode surface, this sensing strategy holds the potential to provide a universal platform for the detection of different metabolites.https://www.mdpi.com/2079-6374/15/5/332enzyme sensorelectrochemiluminescenceimmobilized emitternanochannel arrayglucose |
| spellingShingle | Chunyin Wei Yanyan Zheng Fei Yan Lifang Xu Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection Biosensors enzyme sensor electrochemiluminescence immobilized emitter nanochannel array glucose |
| title | Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection |
| title_full | Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection |
| title_fullStr | Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection |
| title_full_unstemmed | Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection |
| title_short | Enzyme-Based Solid-Phase Electrochemiluminescence Sensors with Stable, Anchored Emitters for Sensitive Glucose Detection |
| title_sort | enzyme based solid phase electrochemiluminescence sensors with stable anchored emitters for sensitive glucose detection |
| topic | enzyme sensor electrochemiluminescence immobilized emitter nanochannel array glucose |
| url | https://www.mdpi.com/2079-6374/15/5/332 |
| work_keys_str_mv | AT chunyinwei enzymebasedsolidphaseelectrochemiluminescencesensorswithstableanchoredemittersforsensitiveglucosedetection AT yanyanzheng enzymebasedsolidphaseelectrochemiluminescencesensorswithstableanchoredemittersforsensitiveglucosedetection AT feiyan enzymebasedsolidphaseelectrochemiluminescencesensorswithstableanchoredemittersforsensitiveglucosedetection AT lifangxu enzymebasedsolidphaseelectrochemiluminescencesensorswithstableanchoredemittersforsensitiveglucosedetection |