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Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates
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Sensing purine nucleoside phosphorylase activity by using silver nanoparticles.

Ya Cao1, Jing Wang, Yuanyuan Xu

  • 1Department of Biochemistry and National Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.

Biosensors & Bioelectronics
|October 15, 2009
PubMed
Summary

This study introduces a novel electrochemical biosensor and UV-vis spectroscopy method for assaying purine nucleoside phosphorylase (PNP) activity. These methods offer reliable, specific, and reagent-free alternatives for clinical applications.

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Area of Science:

  • Biotechnology
  • Electrochemistry
  • Spectroscopy

Background:

  • Purine nucleoside phosphorylase (PNP) is an enzyme crucial in purine metabolism.
  • Accurate and efficient methods for assaying PNP activity are needed for clinical diagnostics.
  • Existing methods often require coupled enzymes or radiochemical reagents, limiting their practicality.

Purpose of the Study:

  • To develop a novel electrochemical biosensor for quantifying PNP activity.
  • To establish a rapid UV-vis spectroscopy assay for PNP activity.
  • To provide reliable, specific, and reagent-free alternatives for PNP activity assessment.

Main Methods:

  • Fabrication of an electrode modified with silver nanoparticles for enhanced electron transfer.
  • Electrochemical detection of guanosine and guanine oxidation, correlated with PNP activity.

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Last Updated: Jun 19, 2026

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  • UV-vis spectroscopy analysis based on the differential agglomeration of silver nanoparticles induced by guanosine and guanine.

    • Main Results:

    • The electrochemical biosensor demonstrated a broad linear range (4-20 unit/mL) and a low detection limit (0.1 unit/mL).
    • The UV-vis spectroscopy method provided a rapid assay for PNP activity.
    • Both methods showed acceptable reliability and specificity, avoiding complex reagents.

    Conclusions:

    • The developed electrochemical biosensor and UV-vis spectroscopy assay are effective tools for determining PNP activity.
    • These novel methods offer advantages in terms of simplicity, cost-effectiveness, and suitability for clinical applications.
    • The findings pave the way for improved diagnostic approaches related to PNP activity.