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A turn-on fluorescence assay of alkaline phosphatase activity using a DNA-silver nanocluster probe

Abstract

Assays of Alkaline phosphatase (ALP) activity play a critical role in clinical diagnostics and drug screening. In this work, we present a novel, sensitive and cost-effective analytical method for the detection of ALP activity based on the design of a G-rich DNA light-up DNA-silver nanocluster (AgNCs) probe and λ exonuclease (exo) cleavage reaction. Upon addition of λ exo, the G-rich DNA was cleaved, resulting in the inhibition of AgNCs to move closer to G-rich DNA sequences and, accordingly, only a low fluorescence signal was observed. Upon treatment of ALP, the 5’-phosphoryl end of pG-rich DNA was hydrolyzed and the λ exo cleavage reaction was impeded. The AgNCs were then enabled to move closer to the G-rich DNA sequences, resulting in an increase in fluorescence. Under optimized conditions, the fluorescence change was determined to be linear with the ALP concentration ranging between 1 U/L and 800 U/L (detection limit: 1 U/L). Taken in concert, this strategy may provide a basis for a screening platform for ALP inhibitors.

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Publication details

The article was received on 11 Dec 2017, accepted on 10 Feb 2018 and first published on 12 Feb 2018


Article type: Paper
DOI: 10.1039/C7NJ04894G
Citation: New J. Chem., 2018, Accepted Manuscript
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    A turn-on fluorescence assay of alkaline phosphatase activity using a DNA-silver nanocluster probe

    C. Ma, H. Liu, K. Wu, M. Chen, H. He, K. Wang and K. Xia, New J. Chem., 2018, Accepted Manuscript , DOI: 10.1039/C7NJ04894G

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