In this work, an electrochemiluminescence (ECL) biosensor based on dual ECL quenching effects of silver nanoclusters (Ag NCs) and multiple cycling amplification was designed to achieve ultrasensitive detection of ATP. The specific recognition of target ATP to aptamer initiated multiple cycling amplification, and a small amount of target was converted into a large number of DNA product chains (S1) by amplification. After S1 opened hairpin DNA 2 (HP2), Ag NCs approached the surface of CdS quantum dots (QDs) modified-electrode by complementary DNA, resulting in a significant decrease of ECL intensity from CdS QDs. The quenching principle is as follows. Firstly, the absorption spectrum of Ag NCs overlaps well with the ECL emission spectrum of CdS QDs, leading to effective ECL resonance energy transfer (ECL-RET); Secondly, Ag NCs could catalyze electrochemical reduction of K2S2O8, leading to consumption of ECL co-reactant and reducing ECL of QDs. The double-ECL quenching achieved ultrasensitive biosensing detection of ATP with a wide range from 1 aM to 1 pM. This present work reported new principle of double-quenching QDs ECL by Ag NCs, and developed a novel ECL biosensor by combining with multiple cycle amplification technique, which has great contribution to the development of QDs ECL and biosensing applications.
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