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

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Abstract

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 (Fe3O4@Au NPs-S1-S2-S3). The magnetic biocomposites were acquired by modifying double-stranded DNA (S1-S2-S3) on the surface of Fe3O4@Au nanoparticles (Fe3O4@Au NPs). Among them, Fe3O4@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.

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Funding

This work was supported by the National Natural Science Foundation of China (Nos. 81773681, 81572081, 21904069) and the Natural Science Foundation of Jiangsu Province (No. BK20190653). This work was also supported by China Postdoctoral Science Foundation (Nos. 2019M661777, 2017M621789) and the Postdoctoral Science Foundation of Jiangsu (Nos. 2019K068, 2019Z176).

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Authors and Affiliations

  1. School of Pharmacy, Nanjing Medical University, Nanjing, 211166, Jiangsu, China

    Xiaoyun Li, Chenyuan Weng, Jing Wang, Wei Yang, Qiaoyun Lu, Xiaoqiang Yan, Marwan Ahmad Sakran, Junli Hong, Wanying Zhu & Xuemin Zhou

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  1. Xiaoyun Li
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  2. Chenyuan Weng
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  3. Jing Wang
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  4. Wei Yang
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Correspondence to Wanying Zhu or Xuemin Zhou.

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Li, X., Weng, C., Wang, J. et al. A label-free electrochemical magnetic aptasensor based on exonuclease III–assisted signal amplification for determination of carcinoembryonic antigen. Microchim Acta 187, 492 (2020). https://doi.org/10.1007/s00604-020-04457-7

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