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Label-free detection of DNA-binding proteins based on microfluidic solid-state molecular beacon sensor.

Jun Wang1, Daisuke Onoshima, Michihiko Aki

  • 1Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Japan. wjzyy988@gmail.com

Analytical Chemistry
|April 12, 2011
PubMed
Summary

This study presents a novel optical sensor for detecting proteins like single-stranded DNA binding protein (SSBP) and histone. The sensor utilizes a gold-supported molecular beacon and microfluidics for sensitive, label-free protein detection.

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Materials Science

Background:

  • Protein detection is crucial for diagnostics and research.
  • Existing methods often require labels or complex procedures.
  • Developing sensitive, label-free detection methods is a key goal.

Purpose of the Study:

  • To develop a novel solid-state optical sensor for label-free detection of specific proteins.
  • To utilize a gold-supported molecular beacon integrated with microfluidics.
  • To demonstrate the sensor's capability for detecting single-stranded DNA binding protein (SSBP) and histone protein.

Main Methods:

  • Fabrication of a solid-state molecular beacon sensor on a gold surface.
  • Integration of the sensor with a polydimethylsiloxane (PDMS) microfluidic channel.
  • Utilizing differential fluorescence quenching of immobilized DNA probes (single-stranded DNA-Cy3 for SSBP, double-stranded DNA-Cy3 for histone) by the gold support.
  • Quantifying protein amounts based on fluorescence recovery or quenching.

Main Results:

  • Achieved label-free detection of SSBP and histone proteins.
  • Demonstrated detection at nanomolar concentrations.
  • The sensor operates in a convenient, general, continuous flow format.
  • Showcased distinct fluorescence responses based on protein binding and DNA probe conformation.

Conclusions:

  • The developed optical sensor offers a sensitive and label-free method for protein detection.
  • The approach leverages binding-induced conformational changes in DNA probes for signal generation.
  • This technology holds significant potential for detecting various proteins with high sensitivity.