Fabrication of a ratiometric electrochemiluminescence biosensor using single self-enhanced nanoluminophores for the detection of spermine

A ratiometric electrochemiluminescence strategy using a single luminophore for accurate and sensitive biomolecule detection could be immensely valuable in bioanalysis. Herein, an ultrasensitive ratiometric electrochemiluminescence sensing system was fabricated using a self-enhanced luminophore with dual-signal emission for the detection of spermine. A nanocomposite was synthesized by the covalent attachment of N, N-diisopropylethylenediamine onto glutathione-protected Au-Ag bimetallic nanoclusters (DPEA-GSH@Au/Ag BNCs). The nanocomposite exhibited efficient intra-cluster charge transfer to produce strong anodic self-enhanced electrochemiluminescence emission at 0.8 V without external co-reactants. Interestingly, the DPEA@GSH@Au-Ag BNCs exhibited cathodic electrochemiluminescence emission upon the addition of the co-reactant potassium persulfate at -1.6 V, exhibiting stable and efficient dual-signal electrochemiluminescence emission features at a continuous potential window of -1.75 to 1.2 V. Thus, they were used to fabricate a single-luminophore electrochemiluminescence sensor with dual emission. The cathodic emission of the biosensor gradually increased with increasing concentrations of spermine, whereas the anodic electrochemiluminescence intensity remained almost constant, enabling the ratiometric detection of spermine. The fabricated biosensor, with an internal standard, significantly improved the accuracy and reliability of spermine detection in a wide concentration range of 0.85 pM-100 μM, with a low limit of detection of 0.12 pM (S/N = 3) under optimum conditions. This single-luminophore electrochemiluminescence sensing system could be used for the detection of spermine and could guide the construction of ratiometric electrochemiluminescence sensors in the future.