Imprinted polymer layer for recognizing double-stranded DNA

Area of Science:

• Polymer Chemistry
• Molecular Recognition
• Biotechnology

Background:

• Developing selective methods for double-stranded DNA (dsDNA) detection is crucial for diagnostics.
• Existing methods often lack specificity or require complex procedures.
• Molecular imprinting offers a promising approach for creating tailored recognition materials.

Purpose of the Study:

• To develop a thin polymer layer capable of specifically recognizing double-stranded DNA (dsDNA).
• To utilize 2-vinyl-4,6-diamino-1,3,5-triazine (VDAT) as a functional monomer for DNA imprinting.
• To evaluate the binding specificity and kinetics of the imprinted polymer for a target dsDNA sequence.

Main Methods:

• Synthesized a DNA-imprinted polymer using VDAT, acrylamide, and a crosslinking agent with verotoxin dsDNA as the template.
• Confirmed hydrogen bonding between VDAT and A-T base pairs through dsDNA melting point, NMR, and CD spectra analysis.
• Assessed binding specificity using fluorescently labeled dsDNAs and analyzed binding kinetics via surface plasmon resonance (SPR).

Main Results:

• The VDAT-based imprinted polymer demonstrated preferential binding to the template verotoxin dsDNA over non-target dsDNA (oligo(dG)-oligo(dC)).
• SPR analysis revealed a low dissociation constant (KD) of approximately 10(-9) M, indicating high binding affinity.
• The polymer layer showed twice the fluorescence intensity when binding verotoxin dsDNA compared to oligo(dG)-oligo(dC).

Conclusions:

• A novel DNA-imprinted polymer utilizing VDAT has been successfully prepared for specific dsDNA recognition.
• The developed material exhibits high selectivity and affinity for the target verotoxin dsDNA.
• This imprinted polymer holds potential for sensitive and specific dsDNA detection applications.