A label-free electrochemical magnetic aptasensor based on exonuclease III–assisted signal amplification for determination of carcinoembryonic antigen

A novel label-free and exonuclease III (Exo III)–assisted signal amplification electrochemical aptasensor was constructed for the determination of carcinoembryonic antigen (CEA) via magnetic field–induced self-assembly of magnetic biocomposites (Fe 3 O 4 @Au NPs-S1-S2-S3). The magnetic biocomposites were acquired by modifying double-stranded DNA (S1-S2-S3) on the surface of Fe 3 O 4 @Au nanoparticles (Fe 3 O 4 @Au NPs). Among them, Fe 3 O 4 @Au NPs were used as carriers for magnetic separation, thiolated single-stranded DNA (S1) provided signal sequence, CEA aptamer (S2) worked as a recognition element, and complementary strand (S3) was used to form double strands. In the presence of CEA, S2 bonded with CEA competitively; the exposed S1 could not be cleaved since Exo III was inactive against ssDNA. The G-quadruplex/hemin complexes finally formed with the existence of K + , and the high electrochemical signal of G-quadruplex/hemin complexes was recorded by differential pulse voltammetry (DPV) at − 0.6 V. Conversely, in the absence of CEA, dsDNA was cleaved from the 3′ blunt end by Exo III; the disappearance of G-rich sequence blocked the generation of the signal. This method exhibited good selectivity and sensitivity for the determination of CEA; the linear range was from 0.1 to 200 ng mL −1 and the limit of detection was 0.4 pg mL −1 . Graphical abstract