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Nanostructured bioluminescent sensor for rapidly detecting thrombin.

Longyan Chen1, Yige Bao2, John Denstedt2

  • 1Department of Chemical and Biochemical Engineering, Western University, London, Ontario, Canada N6A 5B9.

Biosensors & Bioelectronics
|September 24, 2015
PubMed
Summary

This study introduces a novel bioluminescent nanosensor for rapid, non-invasive detection of thrombin in urine. The sensor shows a linear response and a low detection limit, offering a promising tool for clinical diagnosis.

Keywords:
Bioluminescence resonance energy transfer sensorGold nanoparticleLuciferaseThrombin

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Area of Science:

  • Biochemistry
  • Nanotechnology
  • Biomedical Engineering

Background:

  • Thrombin is crucial in hemostasis and thrombosis.
  • Abnormal thrombin levels are linked to diseases like rheumatoid arthritis and glomerulonephritis.
  • Accurate thrombin detection in bodily fluids is vital for medical diagnosis.

Purpose of the Study:

  • To develop a novel bioluminescent sensor for rapid, non-invasive detection of thrombin in urine.
  • To create a nanosensor utilizing gold nanoparticles and Renilla luciferase for thrombin detection.
  • To establish a sensitive and specific assay for thrombin quantification.

Main Methods:

  • Conjugating gold nanoparticles (Au NPs) with a recombinant Renilla luciferase (pRluc) protein via a thrombin-specific peptide substrate.
  • Utilizing the quenching of luciferase bioluminescence by peptide-conjugating Au NPs.
  • Observing bioluminescence recovery upon thrombin-induced peptide digestion and luciferase release.

Main Results:

  • The bioluminescent sensor demonstrated a significant recovery of luminescence intensity upon thrombin digestion within 10 minutes.
  • A linear relationship was observed between luminescence intensity and thrombin concentration (8 nM to 8 μM) in buffer and urine samples.
  • The sensor achieved a low limit of detection of 80 pM for thrombin.

Conclusions:

  • The developed nanosensor enables rapid and non-invasive detection of thrombin in urine.
  • The sensor exhibits high sensitivity and a wide linear detection range, suitable for clinical applications.
  • This nanosensor presents a promising tool for the clinical diagnosis of thrombin-related conditions.