Microchimica Acta

, 185:231 | Cite as

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

Original Paper
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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.

Graphical abstract

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.

Keywords

Polymer quantum dots Fluorescent polymer Ascorbic acid 2-phosphate Bioassay Cell lysates Inhibitor screening 

Notes

Acknowledgements

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

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_2769_MOESM1_ESM.doc (3.7 mb)
ESM 1 (DOC 3748 kb)

References

  1. 1.
    Wu C, Bull B, Szymanski C, Christensen K, McNeill J (2008) Multicolor conjugated polymer dots for biological fluorescence imaging. ACS Nano 2:2415–2423CrossRefGoogle Scholar
  2. 2.
    Ye F, Wu C, Jin Y, Chan YH, Zhang X, Chiu DT (2011) Ratiometric temperature sensing with semiconducting polymer dots. J Am Chem Soc 133:8146–8149CrossRefGoogle Scholar
  3. 3.
    Yu JB, Rong Y, Kuo CT, Zhou XH, Chiu DT (2017) Recent advances in the development of highly luminescent semiconducting polymer dots and nanoparticles for biological imaging and medicine. Anal Chem 89:42–56CrossRefGoogle Scholar
  4. 4.
    Wu CF, Schneider T, Zeigler M, Yu JB, Schiro PG, Burnham DR, McNeill JD, Chiu DT (2010) Bioconjugation of ultrabright semiconducting polymer dots for specific cellular targeting. J Am Chem Soc 132:15410–15417CrossRefGoogle Scholar
  5. 5.
    Massey M, Wu M, Conroy EM, Algar WR (2015) Bioconjugation of ultrabright semiconducting polymer dots for specific cellular targeting. Curr Opin Biotechnol 34:30–40CrossRefGoogle Scholar
  6. 6.
    Liou SY, Ke CS, Chen JH, Luo YW, Kuo SY, Chen YH, Fang CC, Wu CY, Chiang CM, Chan YH (2016) Tuning the emission of semiconducting polymer dots from green to near-infrared by alternating donor monomers and their applications for in vivo biological imaging. ACS Macro Lett 5:154–157CrossRefGoogle Scholar
  7. 7.
    Lee H, Dellatore SM, Miller WM, Messersmith PB (2007) Mussel-inspired surface chemistry for multifunctional coatings. Science 318:426–430CrossRefGoogle Scholar
  8. 8.
    Zhang X, Wang S, Xu L, Feng L, Ji Y, Tao L, Li S, Wei Y (2012) Biocompatible polydopamine fluorescent organic nanoparticles: facile preparation and cell imaging. Nano 4:5581Google Scholar
  9. 9.
    Yildirim A, Bayindir M (2014) Turn-on fluorescent dopamine sensing based on in situ formation of visible light emitting polydopamine nanoparticles. Anal Chem 86:5508–5512CrossRefGoogle Scholar
  10. 10.
    Kong XJ, Wu S, Chen TT, Yu RQ, Chu X (2016) MnO2-induced synthesis of fluorescent polydopamine nanoparticles for reduced glutathione sensing in human whole blood. Nano 8:15604–15610Google Scholar
  11. 11.
    Liu B, Han X, Liu J (2016) Iron oxide nanozyme catalyzed synthesis of fluorescent polydopamine for light-up Zn 2+ detection. Nano 8(28):13620–13626Google Scholar
  12. 12.
    Ding P, Wang H, Song B, Ji X, Su Y, He Y (2017) In situ live-cell nucleus fluorescence labeling with bioinspired fluorescent probes. Anal Chem 89:7861–7868CrossRefGoogle Scholar
  13. 13.
    Coleman JE (1992) Structure and mechanism of alkaline phosphatase. Annu Rev Biophys Biomol Struct 21:441–483CrossRefGoogle Scholar
  14. 14.
    Couttenye MM, Haese PCD, Van Hoof VO, Lemoniatou E, Goodman W, Verpooten GA, Broe MED (1996) Low serum levels of alkaline phosphatase of bone origin: a good marker of adynamic bone disease in haemodialysis patients. Nephrol Dial Transplant 11:1065–1072CrossRefGoogle Scholar
  15. 15.
    Lorente JA, Valenzuela H, Morote J, Gelabert A (1999) Serum bone alkaline phosphatase levels enhance the clinical utility of prostate specific antigen in the staging of newly diagnosed prostate cancer patients. Eur J Nucl Med Mol Imaging 26:625CrossRefGoogle Scholar
  16. 16.
    Kokado A, Arakawa H, Maeda M (2000) New electrochemical assay of alkaline phosphatase using ascorbic acid 2-phosphate and its application to enzyme immunoassay. Anal Chim Acta 407:119–125CrossRefGoogle Scholar
  17. 17.
    Gao Z, Deng K, Wang XD, Miró M, Tang D (2014) High-resolution colorimetric assay for rapid visual readout of phosphatase activity based on gold/silver core/shell nanorod. ACS Appl Mater Interfaces 6:18243–18250CrossRefGoogle Scholar
  18. 18.
    Sun J, Hu T, Chen C, Zhao D, Yang F, Yang X (2016) Fluorescence immunoassay system via enzyme-enabled in situ synthesis of fluorescent silicon nanoparticles. Anal Chem 88:9789–9795CrossRefGoogle Scholar
  19. 19.
    Suzuki K, Kobayashi A, Kaneko S, Takehira K, Yoshihara T, Ishida H, Shiina Y, Oishi S, Tobita S (2009) Reevaluation of absolute luminescence quantum yields of standard solutions using a spectrometer with an integrating sphere and a back-thinned CCD detector. Phys Chem Chem Phys 11:9850–9860CrossRefGoogle Scholar
  20. 20.
    Shamsipur M, Shanehasz M, Khajeh K, Mollania N, Kazemi SH (2012) A novel quantum dot–laccase hybrid nanobiosensor for low level determination of dopamine. Analyst 137:5553–5559CrossRefGoogle Scholar
  21. 21.
    Carver JC, Schweitzer GK, Carlson TA (1972) Use of X-Ray photoelectron spectroscopy to study bonding in Cr, Mn, Fe, and Co compounds. J Chem Phys 57:973CrossRefGoogle Scholar
  22. 22.
    Nicolas RG, Pelletier ML, Nair MM, Chevallier P, Jean Lagueux J, Gossuin Y, Laurent S, Kleitz F, Fortin MA (2013) Manganese-impregnated mesoporous silica nanoparticles for signal enhancement in MRI cell labelling studies. Nano 5:11499Google Scholar
  23. 23.
    Di Castro V, Polzonetti G (1989) XPS study of MnO oxidation. J Electron Spectrosc Relat Phenom 48:117CrossRefGoogle Scholar
  24. 24.
    Jia HL, Cheng JY, Ya CY, Wei LT (2015) Formation of fluorescent polydopamine dots from hydroxyl radical-induced degradation of polydopamine nanoparticles. Phys Chem Chem Phys 17:15124CrossRefGoogle Scholar
  25. 25.
    Yu X, Fan H, Liu Y, Shi Z, Jin Z (2014) Characterization of carbonized polydopamine nanoparticles suggests ordered supramolecular structure of polydopamine. Langmuir 30:5497–5505CrossRefGoogle Scholar
  26. 26.
    Ma JL, Yin BC, Wu X, Ye BC (2016) Copper-mediated DNAscaffolded silver nanocluster on-off switch for detection of pyrophosphate and alkaline phosphatase. Anal Chem 88(18):9219–9225CrossRefGoogle Scholar
  27. 27.
    Kang W, Ding Y, Zhou H, Liao Q, Yang X, Yang Y, Jiang JS, Yang MH, Yang MH (2015) Monitoring the activity and inhibition of alkaline phosphatase via quenching and restoration of the fluorescence of carbon dots. Microchim Acta 182(5–6):1161–1167CrossRefGoogle Scholar
  28. 28.
    Nie F, Luo K, Zheng X, Zheng J, Song Z (2015) Novel preparation and electrochemiluminescence application of luminol functional-Au nanoclusters for ALP determination. Sensors Actuators B Chem 218:152–159CrossRefGoogle Scholar
  29. 29.
    Tang Z, Zhang H, Ma CB, Gu P, Zhang GH, Wu KF, Chen MJ, Wang KM (2018) Colorimetric determination of the activity of alkaline phosphatase based on the use of Cu (II)-modulated G-quadruplex-based DNAzymes. Microchim Acta 185(2):109CrossRefGoogle Scholar
  30. 30.
    Mao M, Tian T, He Y, Ge Y, Zhou J, Song G (2018) Inner filter effect based fluorometric determination of the activity of alkaline phosphatase by using carbon dots codoped with boron and nitrogen. Microchim Acta 185(1):17CrossRefGoogle Scholar
  31. 31.
    Wang HB, Li Y, Chen Y, Zhang ZP, Gan T, Liu YM (2018) Determination of the activity of alkaline phosphatase by using nanoclusters composed of flower like cobalt oxyhydroxide and copper nanoclusters as fluorescent probes. Microchim Acta 185(2):102CrossRefGoogle Scholar
  32. 32.
    Guo L, Chen D, Yang M (2017) DNA templated silver nanoclusters for fluorometric determination of the activity and inhibition of alkaline phosphatase. Microchim Acta 184(7):2165–2170CrossRefGoogle Scholar
  33. 33.
    Liu H, Ma C, Wang J, Wang K, Wu K (2017) A turn-on fluorescent method for determination of the activity of alkaline phosphatase based on dsDNA templated copper nanoparticles and exonuclease based amplification. Microchim Acta 184(7):2483–2488CrossRefGoogle Scholar
  34. 34.
    Wang H, Mu L, She G, Xu H, Shi W (2014) Fluorescent biosensor for alkaline phosphatase based on fluorescein derivatives modified silicon nanowires. Sensors Actuators B Chem 203:774–781CrossRefGoogle Scholar
  35. 35.
    He Y, Jiao B (2017) Determination of the activity of alkaline phosphatase based on the use of ssDNA templated fluorescent silver nanoclusters and on enzyme-triggered silver reduction. Microchim Acta 184(10):4167–4173CrossRefGoogle Scholar
  36. 36.
    Qian ZS, Chai LJ, Huang YY, Tang C, Shen J, Jia C, Feng H (2015) A real-time fluorescent assay for the detection of alkaline phosphatase activity based on carbon quantum dots. Biosens Bioelectron 68:675–680CrossRefGoogle Scholar
  37. 37.
    Dong L, Miao Q, Hai Z, Yuan Y, Liang G (2015) Enzymatic hydrogelation-induced fluorescence turn-off for sensing alkaline phosphatase in vitro and in living cells. Anal Chem 87(13):6475–6478CrossRefGoogle Scholar
  38. 38.
    Deng J, Yu P, Wang Y, Mao L (2015) Real-time ratiometric fluorescent assay for alkaline phosphatase activity with stimulus responsive infinite coordination polymer nanoparticles. Anal Chem 87(5):3080–3086CrossRefGoogle Scholar
  39. 39.
    Dhariwal KR, Hartzell WO, Levine M (1991) Ascorbic acid and dehydroascorbic acid measurements in human plasma and serum. Am J ClinNutr 54:712–716Google Scholar
  40. 40.
    Cameron E, Pauling L, Leibovitz B (1979) Ascorbic acid and cancer: a review. Cancer Res 39:663–681Google Scholar
  41. 41.
    Meister A, Anderson ME (1983) Glutathione. Annu Rev Biochem 52:711–760CrossRefGoogle Scholar
  42. 42.
    Gibbons IR, Cosson MP, Evans JA, Gibbons BH, Houck B, Martinson KH, Sale WS, Tang WJ (1978) Potent inhibition of dynein adenosine triphosphatase and of the motility of cilia and sperm flagella by vanadate. Proc Natl Acad Sci 75:2220–2224CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Shanghai Key Laboratory of Green Chemistry and Chemical Processes, School of Chemistry and Molecular EngineeringEast China Normal UniversityShanghaiChina

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