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Author Spotlight: Advancements in DNA Nanosensors &#8211; Addressing Sensitivity and Selectivity Challenges in Molecular Detection
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DNA immobilization and detection using DNA binding proteins.

Gülsen Betül Aktas1, Arnau Ribera1, Vasso Skouridou1

  • 1Departament d'Enginyeria Química, Universitat Rovira i Virgili, 26 Països Catalans, 43007, Tarragona, Spain.

Analytical and Bioanalytical Chemistry
|January 27, 2021
PubMed
Summary

DNA binding proteins offer a cost-effective alternative to streptavidin-biotin for biosensor immobilization. This novel approach enhances DNA detection signals and simplifies assay development for improved biosensor performance.

Keywords:
DNA immobilizationMicroplateNitrocelluloseProtein adsorptionStreptavidinscCro DNA binding protein

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biosensor Technology

Background:

  • Biosensor performance relies heavily on bioreceptor immobilization.
  • The streptavidin-biotin system is commonly used for DNA immobilization in biosensors.
  • This system requires post-synthesis modification of DNA, increasing costs.

Purpose of the Study:

  • To explore DNA binding proteins as universal anchors for DNA immobilization and detection.
  • To evaluate scCro DNA binding protein as an alternative to streptavidin-biotin.
  • To develop a nucleic acid lateral flow assay using DNA binding proteins.

Main Methods:

  • Immobilization of scCro DNA binding protein on microplates and nitrocellulose membranes.
  • Detection of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) targets.
  • Development of a lateral flow assay using scCro and dHP DNA binding proteins for PCR amplicon detection.

Main Results:

  • scCro enabled successful detection of both ssDNA and dsDNA targets.
  • The scCro system achieved detection limits comparable to streptavidin-biotin but with higher signals and lower protein usage.
  • scCro demonstrated superior adsorption properties compared to streptavidin.
  • A functional nucleic acid lateral flow assay was developed using two DNA binding proteins.

Conclusions:

  • DNA binding proteins, such as scCro, are viable alternatives to streptavidin-biotin for DNA immobilization in biosensors.
  • The proposed system offers potential for cost reduction, improved signal, and simplified assay development.
  • Further research is needed to optimize adsorption on nitrocellulose and enhance assay sensitivity for multiplex detection.