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Amplified electrochemical immunoassay for 5-methylcytosine using a nanocomposite prepared from graphene oxide, magnetite nanoparticles and β-cyclodextrin

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Abstract

A nanocomposite was prepared from β-cyclodextrin (β-CD) functionalized graphene oxide and magnetic nanoparticles (GO/Fe3O4/β-CD). In parallel, a polyamidoamine dendrimer conjugated to avidinylated alkaline phosphatase (PAMAM-avidin-ALP) was prepared and exploited as a signal amplification unit in a voltammetric immunoassay for 5-methylcytosine (5mC) in genomic DNA. The GO/Fe3O4/β-CD as a substrate material exhibited good solubility, electrical conductivity and large surface. This is beneficial for the further modification of antibodies (Ab) by host-guest interaction and amide bonds. By taking advantage of three-dimensional structure to capture avidin-ALP by amide linkages, PAMAM was used as a catalytic signal amplification element in this assay. Under the optimized condition and at a typical working potential of 0.94 V, the response to 5mC is linear in the 0.01–50 nM concentration range with a detection limit of 3.2 pM (at S/N = 3). The method is stable, selective and reproducible. It was applied to the determination of 5mC in genomic DNA of human tissue.

An electrochemical immunoassay was constructed for 5-methylcytosine detection based on nanocomposite of graphene oxide, magnetite nanoparticles and β-cyclodextrin, and enzymatic signal amplification.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 21775090).

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Authors and Affiliations

  1. College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong, 271018, People’s Republic of China

    Yunlei Zhou, Wenjing Jiang, Huanshun Yin, Nan Lu & Shiyun Ai

  2. College of Chemistry and Chemical Engineering, Central South University, 410083, Changsha, Hunan, People’s Republic of China

    Hanwen Wu

  3. The tumor center of Taian city central hospital, 271000, Taian, Shandong, People’s Republic of China

    Fei Liu

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  1. Yunlei Zhou
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Correspondence to Huanshun Yin.

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Zhou, Y., Jiang, W., Wu, H. et al. Amplified electrochemical immunoassay for 5-methylcytosine using a nanocomposite prepared from graphene oxide, magnetite nanoparticles and β-cyclodextrin. Microchim Acta 186, 488 (2019). https://doi.org/10.1007/s00604-019-3575-6

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