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Related Experiment Video

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A plug-and-play aptamer diagnostic platform based on linear dichroism spectroscopy.

Haydn A Little1,2, Aysha Ali1,2, Jake G Carter1,2

  • 1School of Chemistry, University of Birmingham, Birmingham, United Kingdom.

Frontiers in Chemistry
|May 25, 2023
PubMed
Summary

This study introduces a novel aptamer-based assay using M13 bacteriophages and linear dichroism (LD) spectroscopy for sensitive molecular detection. The plug-and-play platform effectively detects the protein thrombin at picomolar levels without labels.

Keywords:
DNAM13aptamerbionanoparticlediagnosticssandwich assay

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Area of Science:

  • Biotechnology
  • Spectroscopy
  • Molecular Biology

Background:

  • Aptamer-based assays offer specific molecular recognition capabilities.
  • Linear dichroism (LD) spectroscopy provides sensitive detection through molecular alignment.
  • Filamentous bacteriophage M13 serves as a scaffold for signal amplification in biosensing.

Purpose of the Study:

  • To develop a plug-and-play sandwich assay platform for aptamer-based detection.
  • To utilize linear dichroism (LD) spectroscopy as a readout method for molecular targets.
  • To demonstrate the platform's efficacy in detecting the protein thrombin.

Main Methods:

  • Bioconjugation of a DNA linker onto M13 bacteriophage backbone.
  • Assembly of aptamer-functionalized M13 bacteriophages via complementary base pairing.
  • Characterization of aptamer secondary structure using circular dichroism (CD) spectroscopy.
  • Confirmation of target binding using fluorescence anisotropy measurements.
  • Detection of thrombin using the developed LD spectroscopy-based sandwich assay.

Main Results:

  • The M13 bacteriophage scaffold facilitated strong LD signals due to its alignment in linear flow.
  • The plug-and-play system successfully generated aptamer-functionalized M13 bacteriophages.
  • The aptamer-based sandwich assay demonstrated high sensitivity for thrombin detection, reaching picomolar (pM) levels.
  • The assay functions as a label-free, homogenous detection system.

Conclusions:

  • The developed plug-and-play aptamer assay using M13 bacteriophages and LD spectroscopy is a highly effective platform for molecular detection.
  • This system shows significant potential as a sensitive, label-free, and homogenous biosensing method.
  • The findings highlight the versatility of M13 bacteriophages in constructing advanced aptamer-based biosensors.