期刊
MICROCHIMICA ACTA
卷 186, 期 8, 页码 -出版社
SPRINGER WIEN
DOI: 10.1007/s00604-019-3575-6
关键词
Voltammetric immunoassay; PAMAM dendrimer; DNA methylation; Phos-tag-biotin; Avidin; Alkaline phosphatase; Genomic DNA; Cancer; p-Nitrophenyl phosphate
资金
- National Natural Science Foundation of China [21775090]
A nanocomposite was prepared from beta-cyclodextrin (beta-CD) functionalized graphene oxide and magnetic nanoparticles (GO/Fe3O4/beta-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/beta-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 0.94V, the response to 5mC is linear in the 0.01-50nM concentration range with a detection limit of 3.2 pM (atS/N=3). The method is stable, selective and reproducible. It was applied to the determination of5mC in genomic DNA of human tissue.
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