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Related Experiment Videos

Aptameric enzyme subunit for biosensing based on enzymatic activity measurement.

Wataru Yoshida1, Koji Sode, Kazunori Ikebukuro

  • 1Department of Biotechnology and Life Science, Tokyo University of Agriculture & Technology, 2-24-16 Naka-machi, Koganei, Tokyo 184-8588, Japan.

Analytical Chemistry
|May 13, 2006
PubMed
Summary

A novel aptameric enzyme subunit (AES) enables sensitive detection of target molecules like adenosine. This biosensor uses enzyme activity changes in a homogeneous solution, simplifying detection without separation.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biosensor Development

Background:

  • Biosensing typically requires target molecule-binding aptamers and enzyme-inhibiting aptamers.
  • The G-quartet structure of thrombin-inhibiting aptamers is crucial for their inhibitory function.
  • Existing biosensing methods can be complex, often requiring separation steps.

Purpose of the Study:

  • To develop a novel aptameric enzyme subunit (AES) for homogeneous biosensing.
  • To create a biosensor capable of detecting target molecules by measuring changes in enzyme activity.
  • To design a universal AES structure adaptable for various target molecule-binding aptamers.

Main Methods:

  • Constructed an aptameric enzyme subunit (AES) by integrating an adenosine-binding aptamer into a thrombin-inhibiting aptamer's G-quartet structure.

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  • Utilized the conformational change of the G-quartet upon adenosine recognition to alter thrombin inhibition activity.
  • Measured thrombin activity in a homogeneous solution to detect adenosine without requiring bound/free separation.

    • Main Results:

    • Successfully detected adenosine by monitoring changes in thrombin activity.
    • Demonstrated that the conformational change in the G-quartet structure directly impacts enzyme inhibition.
    • Developed two types of AES, including a universal design for versatile aptamer integration.

    Conclusions:

    • AES enables simple, high-sensitivity detection of target molecules in homogeneous assays.
    • The developed biosensor eliminates the need for separation steps, streamlining the detection process.
    • The universal AES design offers a flexible platform for developing diverse biosensors.