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Development of a simple, cost-effective and sensitive biosensing strategy is highly desirable to advance the applications in Alzheimer's disease diagnosis. In this paper, we present a simple, label-free and signal-on electrochemiluminescence (ECL) aptasensor for the detection of amyloid-beta (Aβ) peptide using luminol as ECL emitter and in-situ generated reactive oxygen species (ROS) as coreactant via catalytic reaction between Cu2+-Aβ and the dissolved O 2 in the presence of ascorbic acid (AA). Aβ 16 , the binding site of Cu2+ in the monomeric full-length Aβ, was used as a model in present study. As a result, this signal-on ECL aptasensor has exhibited favorable analytical performance for Aβ 16 monomer with a linear range of 1.0 × 10-13 mol/L-1.0 × 10-8 mol/L and a limit of detection of 3.5 × 10-14 mol/L (S/N=3). Furthermore, the proposed biosensor was also able to detect the full length Aβ 40 not only in the phosphate buffer saline (PBS) solution but also in human serum. The presented biosensor represents a promising, simple, turn-on and label-free diagnostic tool for blood analysis. • A simple, label-free and signal-on ECL aptasensor for the detection of Aβ peptide was development. • The aptasensor can monitor the aggregation process of Aβ 16 monomer accelerated by Cu2+. • The proposed biosensor was able to detect the full length Aβ 40 not only in the PBS solution but also in human serum. [ABSTRACT FROM AUTHOR]