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A highly sensitive electrochemical biosensor for protein based on a tetrahedral DNA probe, N- and P-co-doped graphene, and rolling circle amplification


A tetrahedral DNA probe can effectively overcome the steric effects of a single-stranded probe to obtain well-controlled density and minimize nonspecific adsorption. Herein, a highly sensitive electrochemical biosensor is fabricated for determination of protein using a tetrahedral DNA probe and rolling circle amplification (RCA). N- and P-co-doped graphene (NP-rGO) is prepared, and AuNPs are then electrodeposited on it for DNA probe immobilization. Benefitting from the synergistic effects of the excellent electrical conductivity of NP-rGO, the stability of the tetrahedral DNA probe and the signal amplification of RCA, the biosensor achieves a low limit of 3.53 × 10−14 M for thrombin and a wide linear range from 1 × 10−13 to 1 × 10−7 M. This study provides a sensitive and effective method for the detection of protein in peripheral biofluids, and paves the way for future clinical diagnostics and treatment of disease.

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This work was supported by the National Natural Science Foundation of China (21475115), Henan Provincial Science and Technology Innovation Team (C20150026) and the Nanhu Scholars Program of Xinyang Normal University.

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Correspondence to Ke-Jing Huang.

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Wu, Z., Xie, X., Guo, H. et al. A highly sensitive electrochemical biosensor for protein based on a tetrahedral DNA probe, N- and P-co-doped graphene, and rolling circle amplification. Anal Bioanal Chem 412, 915–922 (2020). https://doi.org/10.1007/s00216-019-02314-y

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  • Tetrahedral DNA probe
  • N- and P-Co-doped graphene
  • Rolling circle amplification
  • Protein
  • Sensitive determination