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Detection of mutant p53 using field-effect transistor biosensor.

Sang Hee Han1, Sang Kyu Kim, Kyoungsook Park

  • 1BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-333, Republic of Korea.

Analytica Chimica Acta
|April 13, 2010
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Summary

This study demonstrates a novel field-effect transistor (FET)-type biosensor for detecting mutant p53 protein interactions with DNA. The biosensor successfully monitored sequence-specific DNA-protein binding, offering a promising tool for disease monitoring.

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Area of Science:

  • Biomolecular Engineering
  • Biosensor Technology
  • Molecular Diagnostics

Background:

  • The p53 protein plays a crucial role in tumor suppression.
  • Mutations in p53 are common in various cancers and often lead to altered DNA-binding activity.
  • Accurate detection of mutant p53 DNA-binding is essential for cancer diagnosis and monitoring.

Purpose of the Study:

  • To develop and evaluate a novel field-effect transistor (FET)-type biosensor for detecting DNA-protein interactions.
  • To assess the DNA-binding capabilities of wild-type p53 and a specific mutant p53 (R248W) using this biosensor.
  • To establish a method for monitoring mutant p53 based on its DNA-binding activity.

Main Methods:

  • Construction of the core domain of wild-type and mutant p53 (R248W) proteins.
  • Immobilization of consensus DNA-binding sequences onto the sensing layer of a MOSFET biosensor.
  • Application of p53 proteins to the DNA-coated gate surface and analysis using a semiconductor analyzer.
  • Validation of results using surface plasmon resonance (SPR) measurements.

Main Results:

  • The FET-type biosensor successfully detected sequence-specific DNA-protein interactions.
  • Wild-type p53 binding to DNA induced a significant up-shift in drain current.
  • Mutant p53 (R248W), a DNA-binding-defective mutant, did not induce a significant change in drain current.
  • The biosensor results correlated with SPR measurements.

Conclusions:

  • A FET-type biosensor can effectively monitor DNA-protein interactions, specifically for wild-type and mutant p53.
  • This biosensor technology shows promise for detecting mutant p53 based on its DNA-binding activity.
  • The developed method provides valuable insights for disease monitoring, particularly for conditions involving DNA-protein binding events.