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  • Title: Amplified electrochemical immunoassay for 5-methylcytosine using a nanocomposite prepared from graphene oxide, magnetite nanoparticles and β-cyclodextrin.
    Author: Zhou Y, Jiang W, Wu H, Liu F, Yin H, Lu N, Ai S.
    Journal: Mikrochim Acta; 2019 Jul 02; 186(8):488. PubMed ID: 31267252.
    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. Graphical abstract 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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