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An Immunosensing System Using Stilbene Glycoside as a Fluorogenic Substrate for an Enzymatic Reaction Model.

Xue-Hui Zhan1, Fu-Chun Gong2, Shu-Zhen Tan1

  • 1College of Chemistry and Biological Engineering, Changsha University of Science and Technology, Changsha, 410076, Hunan, P.R. China.

Sensors (Basel, Switzerland)
|November 23, 2016
PubMed
Summary

Stilbene glycoside (TBG) is a novel fluorogenic substrate for horseradish peroxidase (HRP) detection. This natural product shows potential in enzyme-linked immunosensing systems for detecting Brucella melitensis antibodies.

Keywords:
Brucella melitensisHRP fluorogenic substrateStilbene glycosideenzyme-linked immunosensing

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biosensors

Background:

  • Horseradish peroxidase (HRP) is widely used in immunoassays.
  • Development of novel fluorogenic substrates can enhance assay sensitivity and efficiency.
  • Natural products offer potential for new biosensing applications.

Purpose of the Study:

  • To evaluate stilbene glycoside (TBG) as a fluorogenic substrate for HRP.
  • To develop and assess an enzyme-linked immunosensing system using TBG.
  • To compare TBG with existing HRP substrates.

Main Methods:

  • TBG was tested as a substrate for HRP-catalyzed reactions.
  • A competitive immunoassay was designed using HRP-conjugated Brucella melitensis antibody (HRP-BrAb) and immobilized antigen (BrAg).
  • The immunosensing system utilized a graphite-based electrode modified with BrAg and HRP-BrAb.

Main Results:

  • TBG demonstrated effective HRP detection, catalyzing a fluorogenic reaction with H₂O₂.
  • The enzyme-linked immunosensing system showed a linear detection range of 3.5´10⁻⁸~7.6´10⁻⁶ g/L for Brucella melitensis antibody.
  • A low detection limit of 1.7´10⁻⁹ g/L was achieved, with good reproducibility (RSD=3.8%) and regenerable sensing surface.

Conclusions:

  • Stilbene glycoside (TBG) is a viable and effective fluorogenic substrate for HRP.
  • The developed HRP-based immunosensing system using TBG is sensitive and reproducible.
  • TBG offers a promising alternative for fluorometric enzyme-linked immunosensing applications, including clinical diagnostics.