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Ultrasensitive Biosensing Platform Based on Hydroxylamine-Enhanced Copper-Mediated Fenton-Like Reaction: Application
Tingting Ma1, Yiran Liu1, Maoguo Li1
1The Key Laboratory of Functional Molecular Solids, Ministry of Education, Anhui Province Key Laboratory of Biomedical Materials and Chemical Measurement, College of Chemistry and Materials Science, Anhui Normal University, Wuhu 241000, China.
Abstract:
The burgeoning field of reactive oxygen species (ROS)-based biosensing holds significant promise for advancements in disease diagnosis and therapeutic monitoring. Herein, we describe the innovative development of a hydroxylamine (HA)-mediated copper-based Fenton-like reaction (FLR) system. A key advantage of this approach lies in its ability to function optimally at neutral pH, a critical departure from traditional iron-catalyzed Fenton reactions that are hindered by metal ion hydrolysis. By incorporating HA as a cyclic reductant, we engineered an efficient amplification mechanism [Cu(II)→HA→Cu(I)→H2O2→•OH], amplifying hydroxyl radical (•OH) generation by 5.6-fold. This robust amplification enabled the development of a sensitive copper ion detection assay exhibiting a wide linear range (0.05-100 nM) and an exceptionally low limit of detection (LOD) of 0.01 nM, which was subsequently validated in biological matrices such as human hair. Furthermore, magnetically carboxylated copper-loaded iron oxide nanoparticles (Cu/Fe3O4-COOH) were synthesized for the first time as highly efficient metal carriers, enabling the ultrasensitive detection of nucleic acids. Through a sophisticated assay design incorporating DNA hybridization probes labeled with these nanoparticles and efficient Fe3+ masking by F-, we achieved ultrasensitive and sequence-specific detection of hepatitis B virus (HBV) DNA. The assay displayed a linear response from 0.1 to 10 nM with an LOD of 0.06 nM, and demonstrated excellent performance in clinical serum samples with spiked recoveries ranging from 97.1 to 108.0%. This research not only introduces a simple strategy for metal ion sensing but also paves the way for the rational design of ultrasensitive nucleic acid biosensors. The demonstrated utility of Fe3O4-COOH as a functionalizable nanocarrier opens avenues for its application in diverse biosensing platforms and other fields requiring advanced nanomaterials.
