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Microbial biosensors are analytical devices that utilize living microbes to detect specific substances through measurable signals. These devices consist of two main components: biosensing organisms and signal-transducing elements. Biosensing organisms, such as Escherichia coli or Saccharomyces cerevisiae, are typically housed in multiwell plates connected to transducers, enabling rapid, real-time detection of target analytes.Signal Generation MechanismWhen a target analyte—such as...

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Single-Particle Plasmon Sensor to Monitor Proteolytic Activity in Real Time.

Rui Oliveira-Silva1,2,3, Yuyang Wang1, Sjoerd W Nooteboom1

  • 1MBx Molecular Biosensing, Department of Applied Physics and Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands.

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Summary

This study introduces a label-free plasmonic sensor for real-time monitoring of thrombin activity. The gold nanorod sensor detects low thrombin levels, aiding in bleeding disorder diagnostics and drug studies.

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Area of Science:

  • Biophysics
  • Nanotechnology
  • Biochemistry

Background:

  • Proteolytic activity is crucial in biological processes.
  • Thrombin plays a key role in blood coagulation.
  • Real-time monitoring of enzyme activity is essential for diagnostics.

Purpose of the Study:

  • To develop a label-free plasmonic sensor for real-time monitoring of thrombin proteolytic activity.
  • To characterize the sensor's performance and kinetic parameters.
  • To explore applications in diagnostics and biochemical studies.

Main Methods:

  • Fabrication of a random array of gold nanorods functionalized with a thrombin-cleavable peptide.
  • Real-time monitoring of plasmon shifts in individual nanorods.
  • Kinetic modeling to understand sensor response.

Main Results:

  • The sensor demonstrated a real-time blueshift in plasmon resonance upon thrombin cleavage.
  • Kinetic analysis revealed a competition between peptide cleavage and thrombin binding.
  • The sensor achieved a dynamic range over two orders of magnitude, detecting thrombin down to 3 nM.
  • Physiologically relevant levels of active thrombin were detected.

Conclusions:

  • The developed plasmonic sensor provides a sensitive and label-free method for quantifying thrombin activity.
  • This technology has potential applications in diagnosing bleeding disorders and in pharmacological research.
  • The sensor's ability to monitor enzymatic activity in real-time opens new avenues for biochemical studies.