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Coiled-coil peptide based sensor for ultra-sensitive thrombin detection.

Patthara Kongsuphol1, Sunil K Arya1, Chee Chung Wong1

  • 1Institute of Microelectronics, 11 Science Park Road, Singapore Science Park II, 117685 Singapore, Singapore.

Biosensors & Bioelectronics
|December 21, 2013
PubMed
Summary

A novel electrochemical biosensor using a gold microelectrode array functionalized with peptide was developed for ultrasensitive thrombin detection. This disposable sensor offers high specificity and a low limit of detection for thrombin.

Keywords:
BiosensorCoiled-coil peptideElectrochemical impedanceSelf-assembled monolayerThrombin

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Area of Science:

  • Biomedical Engineering
  • Electrochemistry
  • Biosensor Technology

Background:

  • Thrombin is a critical enzyme in hemostasis and thrombosis, making its accurate detection vital for diagnosing and managing coagulation disorders.
  • Existing methods for thrombin detection often lack the sensitivity, speed, or specificity required for rapid clinical diagnostics.
  • Development of ultrasensitive and selective biosensors is crucial for advancing point-of-care diagnostics and therapeutic monitoring.

Purpose of the Study:

  • To fabricate an ultrasensitive, disposable, electrochemical biosensor for the detection of thrombin.
  • To functionalize a comb structured gold microelectrode array (CSGMA) with a specific peptide for enhanced thrombin capture and detection.
  • To characterize the performance of the developed biosensor, including its limit of detection, response time, and selectivity.

Main Methods:

  • Fabrication of a novel biosensor using a comb structured gold microelectrode array (CSGMA).
  • Functionalization of CSGMA with a self-assembled monolayer of thiol-terminated coiled-coil peptide (CCP) containing a thrombin-specific cleavage site.
  • Characterization using label-free electrochemical impedance spectroscopy (EIS) and testing with varying thrombin concentrations.

Main Results:

  • The developed CCP/CSGMA electrodes demonstrated an ultrasensitive limit of detection (LOD) for thrombin as low as 10 fg/ml (28 fM).
  • The biosensor achieved catalytic activity detection of thrombin within a rapid 30-minute timeframe.
  • High specificity was observed against other IgG antibodies (DO1 and HA), confirming selective thrombin recognition.

Conclusions:

  • The CCP/CSGMA electrochemical biosensor offers a highly sensitive and specific platform for thrombin detection.
  • The disposable nature and rapid detection capability make it suitable for potential clinical applications.
  • The study provides mechanistic insights into thrombin binding and cleavage processes at the electrode surface.