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Polydopamine nanodots are viable probes for fluorometric determination of the activity of alkaline phosphatase via the in situ regulation of a redox reaction triggered by the enzyme

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Abstract

The authors describe an environmentally friendly and fast (~14 min) method for the synthesis of homogeneously distributed fluorescent polydopamine nanodots (PDA-NDs) using KMnO4 as the oxidant. Alkaline phosphatase (ALP) catalyzes the hydrolysis of ascorbic acid 2-phosphate to release free ascorbic acid which undergoes an in-situ redox reaction with KMnO4. Depending on the activity of ALP, more or less KMnO4 is consumed, and this affects the formation of the PDA-NDs. Based on this finding, a sensitive method was worked out to quantify the activity of ALP via real-time formation of fluorescent PDA-NDs. The fluorometric signal (best measured at excitation/emission peaks of 390/500 nm) is linear in the 1 to 50 mU·mL−1 ALP activity range, and the limit of the detection is as low as 0.94 mU·mL−1 (based on 3 σ/m). The method was successfully applied to the determination of ALP activity in spiked human serum and in MCF-7 cell lysates. It was also applied in a method to screen for inhibitors of ALP.

Schematic of a fluorometric method for the determination of alkaline phosphatase (ALP) activity. The method is based on the in-situ regulation of the formation of fluorescent polydopamine nanodots (PDA-NDs) through the competition between the KMnO4-induced polymerization of dopamine and ALP-directed ascorbic acid 2-phosphate (Asc-2P) hydrolysis. AA: Ascorbic acid.

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Acknowledgements

This work was financially supported from the National Natural Science Foundation of China (21605050) and Shanghai Natural Science Foundation (15ZR1411600).

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Authors and Affiliations

  1. Shanghai Key Laboratory of Green Chemistry and Chemical Processes, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai, 20041, China

    Qin Xue, Xuanyu Cao, Cuiling Zhang & Yuezhong Xian

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  1. Qin Xue
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  2. Xuanyu Cao
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Correspondence to Yuezhong Xian.

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Xue, Q., Cao, X., Zhang, C. et al. Polydopamine nanodots are viable probes for fluorometric determination of the activity of alkaline phosphatase via the in situ regulation of a redox reaction triggered by the enzyme. Microchim Acta 185, 231 (2018). https://doi.org/10.1007/s00604-018-2769-7

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