Hexylamine Functionalized Graphene Oxide-Silver Nanocomposite for Electrochemical Monitoring of Biotin-Streptavidin Binding

Abstract

Electrochemical sensors have emerged as a feasible approach for biomarker detection owing to their high sensitivity, operational simplicity, and cost-effectiveness. However, the development of low-cost, convenient, and common substrate materials with high effectiveness remains a significant challenge. This work presents a new kind of hybrid GO/AgNPs nanostructure (Ag@HA-GO) for electrochemical detection of biotin-streptavidin (SA) interactions in biomarkers. Specifically, GO was firstly modified with hexylamine (HA) to expand the interlayer spacing, and then silver nanoparticles (AgNPs) were incorporated within HA-GO sheets, affording Ag@HA-GO with good dispersibility and excellent electrical conductivity. To utilize it as a substrate material for detecting biotin-streptavidin (biotin–SA) interactions, biotin was covalently attached onto the edges of GO sheets within Ag@HA-GO. In subsequent electrochemical detection of biotin-SA interaction, it was informed that GCE/biotin-Ag@HA-GO could serve as a promising substrate for a sensitive and stable electrochemical sensor, on account of systematical cyclic voltammetry curves. Thus, the designed GO/AgNPs hybrid nanocomposite is expected to act as an effective platform for stabilizing AgNPs, while Ag@GO-HA demonstrates good stability, selectivity, and reproducibility in the electrochemical detection of biotin–SA under physiological conditions.

Keywords

electrochemical detection / graphene oxide / nanocomposite / silver nanoparticles

References

[1] J. Therriault, S. E. Schindler, G. Salvadó, et al., “Biomarker-Based Staging of Alzheimer Disease: Rationale and Clinical Applications,” Nature Reviews Neurology 20 (2024): 232–244.

[2] H. Liu, J. Song, Z. Zhao, S. Zhao, Z. Tian, and F. Yan, “Organic Electrochemical Transistors for Biomarker Detections,” Advancement of Science 11 (2024): 2305347.

[3] J. E. Medina, A. V. Annapragada, P. Lof, et al., “Early Detection of Ovarian Cancer Using Cell-Free DNA Fragmentomes and Protein Biomarkers,” Cancer Discovery 15 (2025): 105–118.

[4] T. Špringer, M. Bocková, J. Slabý, F. Sohrabi, M. Čapková, and J. Homola, “Surface Plasmon Resonance Biosensors and Their Medical Applications,” Biosensors and Bioelectronics 278 (2025): 117308.

[5] D. Lu, J. Lin, D. Su, J. Chen, and R. Qiu, “Recent Progress in SERS Technology Applications in Lung Cancer Detection: From Biomarkers to Samples,” Spectrochimica Acta A 343 (2025): 126481.

[6] P. C. Weber, D. H. Ohlendorf, J. J. Wendoloski, and F. R. Salemme, “Structural Origins of High-Affinity Biotin Binding to Streptavidin,” Science 243 (1989): 85–88.

[7] A. J. Haes and R. P. Van Duyne, “A Nanoscale Optical Biosensor: Sensitivity and Selectivity of an Approach Based on the Localized Surface Plasmon Resonance Spectroscopy of Triangular Silver Nanoparticles,” Journal of the American Chemical Society 124 (2002): 10596–10604.

[8] J. Liu, F. Chen, Y. Song, Y. Chen, and F. Zhang, “Construction of a Highly Selective Enrichment, Ionization, and Detection Platform Based on a Broad-Spectrum Antibody,” Analytical Chemistry 95 (2023): 10984–10991.

[9] L. Liu, D. Wu, S. Zhen, K. Lu, X. Yi, and Z. Sun, “Electrochemical Detection of Telomerase in Cancer Cells Based on the In-Situ Formation of Streptavidin-Biotin-DNA-Biotin Networks for Signal Amplification,” Sensors and Actuators B: Chemical 334 (2021): 129659.

[10] B. Liu, S. Shao, J. Cai, et al., “Signal Cascade Amplification of Streptavidin-Biotin-Modified Immunofluorescence Nanocapsules for Ultrasensitive Detection of Glial Fibrillary Acidic Protein,” Chinese Chemical Letters 36 (2025): 109814.

[11] W. Lee, J. Lee, H. Lee, et al., “Quantum Dot Composite Colloids With Layer-By-Layer Shells: Biological Self-Assemblies for Signal Amplified Detection,” ACS Nano 19 (2025): 30810–30822.

[12] X. Liu, M. Wei, E. Xu, et al., “A Sensitive, Label-Free Electrochemical Detection of Telomerase Activity Without Modification or Immobilization,” Biosensors and Bioelectronics 91 (2017): 347–353.

[13] L. Zhang, X.-M. Shi, Y.-T. Xu, et al., “Gold Nanoparticle-Induced Photocurrent Quenching and Recovery of Polymer Dots: Toward Signal-On Energy-Transfer-Based Photocathodic Bioanalysis of Telomerase Activity in Cell Extracts,” Analytical Chemistry 91 (2019): 6403–6407.

[14] H. Xu, N. Chen, S. Zhang, et al., “An In Situ Quenching Photoelectrochemical Biosensor Sensitized With Dual-Amplification DNA Circuit for Telomerase Activity Detection in HeLa Cells,” Sensors and Actuators B: Chemical 447 (2026): 138813.

[15] N. Xia, Z. Sun, F. Ding, Y. Wang, W. Sun, and L. Liu, “Protease Biosensor by Conversion of a Homogeneous Assay Into a Surface-Tethered Electrochemical Analysis Based on Streptavidin–Biotin Interactions,” ACS Sensors 6 (2021): 1166–1173.

[16] G. Li, X. Yu, D. Liu, X. Liu, F. Li, and H. Cui, “Label-Free Electrochemiluminescence Aptasensor for 2,4,6-Trinitrotoluene Based on Bilayer Structure of Luminescence Functionalized Graphene Hybrids,” Analytical Chemistry 87 (2015): 10976–10981.

[17] X. Liu, Q. Zhang, W. Knoll, B. Liedberg, and Y. Wang, “Rational Design of Functional Peptide–Gold Hybrid Nanomaterials for Molecular Interactions,” Advanced Materials 32 (2020): 2000866.

[18] M. La, Y. Zhang, Y. Gao, M. Li, L. Liu, and Y. Chang, “Impedimetric Detection of MicroRNAs by the Signal Amplification of Streptavidin Induced In Situ Formation of Biotin Phenylalanine Nanoparticle Networks,” Journal of the Electrochemical Society 167 (2020): 117505.

[19] J. Wang, X. Cui, L. Liang, J. Li, B. Pang, and J. Li, “Advances in DNA-Based Electrochemical Biosensors for the Detection of Foodborne Pathogenic Bacteria,” Talanta 275 (2024): 126072.

[20] G. Li, X. Qi, J. Wu, et al., “Highly Stable Electrochemical Sensing Platform for the Selective Determination of Pefloxacin in Food Samples Based on a Molecularly Imprinted-Polymer-Coated Gold Nanoparticle/Black Phosphorus Nanocomposite,” Food Chemistry 436 (2024): 137753.

[21] G. Li, J. Wu, X. Qi, et al., “Molecularly Imprinted Polypyrrole Film-Coated Poly(3,4-Ethylenedioxythiophene): Polystyrene Sulfonate-Functionalized Black Phosphorene for the Selective and Robust Detection of Norfloxacin,” Materials Today Chemistry 26 (2022): 101043.

[22] I. Leka Kottaiveedu Sivakumar, L. Bouffier, N. Sojic, and S. Senthil Kumar, “Enhanced Electrochemiluminescence by Knocking Out Gold Active Sites,” Angewandte Chemie International Edition 64 (2025): e202421185.

[23] W. Yang, M. Pan, C. Huang, Z. Zhao, J. Wang, and H. Zeng, “Graphene Oxide-Based Noble-Metal Nanoparticles Composites for Environmental Application,” Composites Communications 24 (2021): 100645.

[24] X. Li and P. Liu, “Solvothermal In-Situ Preparation of Poly(1,5-Diaminoanthraquinone) Grafted Graphene With Superior Electrochemical Performance,” Composites Communications 34 (2022): 101257.

[25] X. Xu, C. Liu, W. Zhang, and X. Zou, “Active Temperature Regulation and Teamed Boronate Affinity-Facilitated Microelectrode Module for Blood Glucose Detection in Physiological Environment,” Sensors and Actuators B: Chemical 324 (2020): 128720.

[26] T. Rasheed and K. Rizwan, “Metal-Organic Frameworks Based Hybrid Nanocomposites as State-of–the-Art Analytical Tools for Electrochemical Sensing Applications,” Biosensors and Bioelectronics 199 (2022): 113867.

[27] N. Murugan, R. Jerome, M. Preethika, A. Sundaramurthy, and A. K. Sundramoorthy, “2D-Titanium Carbide (MXene) Based Selective Electrochemical Sensor for Simultaneous Detection of Ascorbic Acid, Dopamine and Uric Acid,” Journal of Materials Science & Technology 72 (2021): 122–131.

[28] S. Dong, Z. Yang, B. Liu, et al., “(Pd, Au, Ag) Nanoparticles Decorated Well-Ordered Macroporous Carbon for Electrochemical Sensing Applications,” Journal of Electroanalytical Chemistry 897 (2021): 115562.

[29] C. Walgama and N. Raj, “Silver Nanoparticles in Electrochemical Immunosensing and the Emergence of Silver–Gold Galvanic Exchange Detection,” Chemical Communications 59 (2023): 11161–11173.

[30] F. Sun, X. Wang, Z. You, et al., “Sandwich Structure Confined Gold as Highly Sensitive and Stable Electrochemical Non-Enzymatic Glucose Sensor With Low Oxidation Potential,” Journal of Materials Science & Technology 123 (2022): 113–122.

[31] X. Qi, Y. Xia, J. Wu, et al., “Robust Determination of Marbofloxacin Based on Bimetallic Au@Ag Nanoparticle-Decorated Black Phosphorus Nanosheets Supported Molecularly Imprinted Polymer Film,” Materials Today Chemistry 45 (2025): 102687.

[32] J. Wu, Y. Xia, T. Wang, Y. Zhang, and G. Li, “Efficient Voltammetric Platform Combining a Molecularly Imprinted Polymer and Silver-Nanoparticle-Decorated Black Phosphorus Nanosheets for Selective Determination of Gatifloxacin,” Food Chemistry: X 25 (2025): 102094.

[33] G. Balkourani, A. Brouzgou, and P. Tsiakaras, “A Review on Recent Advancements in Electrochemical Detection of Dopamine Using Carbonaceous Nanomaterials,” Carbon 213 (2023): 118281.

[34] Z. Shen, Y. Peng, X. Li, et al., “Design Principle and Synthetic Strategy for Metal-Organic Framework Composites,” Composites Communications 48 (2024): 101933.

[35] G. Li, H. Yuan, J. Mou, et al., “Electrochemical Detection of Nitrate With Carbon Nanofibers and Copper Co-Modified Carbon Fiber Electrodes,” Composites Communications 29 (2022): 101043.

[36] D. Mathivanan, K. S. Shalini Devi, G. Sathiyan, et al., “Novel Polypyrrole-Graphene Oxide-Gold Nanocomposite for High Performance Hydrogen Peroxide Sensing Application,” Sensors and Actuators, A: Physical 328 (2021): 112769.

[37] E. Klekotka, M. Kasztelan, and B. Palys, “Factors Influencing the Electrocatalytic Properties of Graphene Oxide–Gold Nanoparticles Hybrid System,” ChemElectroChem 8 (2021): 3080–3088.

[38] W. Qian, T. Song, M. Ye, X. Huang, Y. Li, and B. Hao, “Au-Covered Nanographene Oxide/PEG/PAMAM for Surface-Enhanced Raman Scattering Detection,” Composites Communications 23 (2021): 100598.

[39] N. Li, J. Song, P. Wei, et al., “Ultrathin Graphene Oxide Sheets Wrapped MoO₂ Nanoparticles With Molecule Enrichment Function for Sensitive SERS Detection,” Analytical Chemistry 97 (2025): 23505–23515.

[40] M. Kasztelan, A. Studzinska, G. Z. Żukowska, and B. Pałys, “Silver–Graphene Oxide Nanohybrids for Highly Sensitive, Stable SERS Platforms,” Frontiers in Chemistry 9 (2021): 665205.

[41] P. Song, Z. Cai, J. Li, et al., “Construction of rGO-MXene@FeNi/Epoxy Composites With Regular Honeycomb Structures for High-Efficiency Electromagnetic Interference Shielding,” Journal of Materials Science & Technology 217 (2025): 311–320.

[42] B. Hao, G. Lu, S. Zhang, Y. Li, A. Ding, and X. Huang, “Gold Nanoparticles Standing on PEG/PAMAM/Thiol-Functionalized Nanographene Oxide as Aqueous Catalysts,” Polymer Chemistry 11 (2020): 4094–4104.

[43] A. Thesing, L. F. Loguercio, B. S. Noremberg, et al., “Tunable Graphene Oxide Inter-Sheet Distance to Obtain Graphene Oxide–Silver Nanoparticle Hybrids,” New Journal of Chemistry 43 (2019): 1285–1290.

[44] O. C. Compton, D. A. Dikin, K. W. Putz, L. C. Brinson, and S. T. Nguyen, “Electrically Conductive “Alkylated” Graphene Paper via Chemical Reduction of Amine-Functionalized Graphene Oxide Paper,” Advanced Materials 22 (2010): 892–896.

[45] S. Ding, H. Hu, X. Yue, et al., “A Fluorescent Biosensor Based on Quantum Dot–Labeled Streptavidin and Poly-l-Lysine for the Rapid Detection of Salmonella in Milk,” Journal of Dairy Science 105 (2022): 2895–2907.

[46] L. Xu, J. Zhao, and Z. J. D. Wang, “Pigments, Photo-Physicochemical Properties and In Vitro Photodynamic Therapy Activity of Morpholine-Substituted Zinc(II)-Phthalocyanines π-π Stacked on Biotinylated Graphene Quantum Dots,” Pigments, Dyes Pigments 165 (2019): 536–545.

[47] P. Kar, F. Tatard, G. Lamblin, et al., “Silver Nanoparticles to Improve Electron Transfer at Interfaces of Gold Electrodes Modified by Biotin or Avidin,” Journal of Electroanalytical Chemistry 692 (2013): 17–25.

[48] S. Han, W. Liu, M. Zheng, and R. Wang, “Label-Free and Ultrasensitive Electrochemical DNA Biosensor Based on Urchinlike Carbon Nanotube-Gold Nanoparticle Nanoclusters,” Analytical Chemistry 92 (2020): 4780–4787.

[49] A. Wang, Y. Li, X. You, et al., “Electrochemical Immunosensor Nanoarchitectonics With the Ag-rGO Nanocomposites for the Detection of Receptor-Binding Domain of SARS-CoV-2 Spike Protein,” Journal of Solid State Electrochemistry 27 (2023): 489–499.

[50] Z. Shao, H. Yang, T. Wang, X. Zeng, and Q. Xie, “Split-Type Photoelectrochemistry Aptasensing of Kanamycin Based on the Split Aptamers and the Fe(III)-Coordination-Regulated Curcumin Sensitization of TiO₂,” Sensors and Actuators B: Chemical 444 (2025): 138444.

RIGHTS & PERMISSIONS

2026 The Author(s). SmartMat published by Tianjin University and John Wiley & Sons Australia, Ltd.