Herein, we reported an efficient electrochemiluminescence (ECL) biosensor chip for sensitive detection of neuron-specific enolase (NSE). First, 3,4,9,10-perylenetetracarboxylic acid with good luminescence characteristics was used as a luminophore to obtain a stable ECL signal. Subsequently, hollow porous Co3O4/CuO concave polyhedron nanocages (CPNCs) were designed as co-reaction promoters to amplify the luminescence signals for highly sensitive trace detection of NSE. In brief, the rapid cyclic conversion of Co3+/Co2+ and Cu2+/Cu+ redox pairs could continuously catalyze the reduction of persulfate (S2O8 2–), thus providing a large number of essential active intermediates (SO4 •–) for ECL emission. Meanwhile, the unique structure of Co3O4/CuO CPNCs possessed a large specific surface area, which greatly improved its catalytic efficiency. Third, NKFRGKYKC was developed as an affinity ligand for specific antibody fixation, which improved incubation efficiency and protected bioactivity of antibodies. Finally, we independently designed a microchip and applied it for ECL detection to improve the practical application ability of the sensor. The developed biosensor exhibited good sensitivity with a wide linear range (10 fg/mL to 100 ng/mL) and a low detection limit (3.42 fg/mL), which played an active role in the clinical application of sensing analysis.

In this work, based on electrochemiluminescence (ECL) technology and self-assembled portable disease detection chips, a bioactivity-maintained sensing platform was developed for the quantitative detection of neuron-specific enolase. First, we prepared Fe3O4@MoS2 nanocomposites as an efficient catalyst to accelerate the reduction of persulfate (S2O8 2–). Specifically, abundant sulfate radicals (SO4 •–) were generated because of cyclic conversion between Fe2+ and Fe3+. Meanwhile, MoS2 nanoflowers with a high specific surface area could not only load more Fe3O4 but also solve its agglomeration problem, which greatly improved the catalytic efficiency. Moreover, a biosensor chip was constructed by standard lithography processes for disease detection, which had good sensitivity and portability. According to the above strategies, the developed portable sensing platform played the part of promoting the practical application of bioanalysis in early tumor screening and clinical diagnosis. In addition, we developed a short peptide ligand (NARKFYKG, NAR) to avoid the occupation of antigen binding sites by specifically connecting to Fc fragments in antibodies. Thus, the binding efficiency of antibodies and the activity of biosensors were improved due to the introduction of NAR.

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