Boosting biomolecular interactions through DNA origami nano-tailored biosensing interfaces

Area of Science:

• Biotechnology
• Nanotechnology
• Materials Science

Background:

• Biosensor performance relies on optimal bioreceptor interaction with targets.
• Uncontrolled bioreceptor deposition leads to suboptimal spacing and orientation, hindering biomolecular interactions and reducing device sensitivity and specificity.
• Developing advanced immobilization strategies is crucial for improving biosensor functionality.

Purpose of the Study:

• To investigate the use of 3D DNA origami for nanotailored bioreceptor immobilization on microparticles.
• To engineer specific anchoring points for controlled bioreceptor arrangement in a microfluidic environment.
• To evaluate the impact of DNA origami-based immobilization on biosensor binding kinetics, efficiency, and overall performance.

Main Methods:

• Fabrication of disc-shaped microparticles with DNA origami structures featuring tailored anchoring points.
• Immobilization of bioreceptors onto the DNA origami nanostructured surface within a continuous microfluidic system.
• Characterization of bioreceptor surface density, orientation, and accessibility.
• Performance evaluation of the DNA origami-based biosensor compared to a non-DNA origami reference system using binding kinetics, efficiency, and aptamer-based sandwich assays.

Main Results:

• The DNA origami immobilization strategy resulted in a less densely packed bioreceptor surface with reduced steric hindrance and favored upward orientation.
• Bioreceptor accessibility was significantly increased, leading to a 4-fold enhancement in binding kinetics and a 6-fold increase in binding efficiency.
• The DNA origami nanotailored biosensing approach demonstrated an 11-fold improvement in the limit of detection and a 2.5-fold improvement in signal-to-noise ratio compared to traditional aptamer coupling.
• The developed system outperformed the reference system in an aptamer-based sandwich bioassay.

Conclusions:

• 3D DNA origami provides a powerful tool for bottom-up nanoscale engineering of biosensor surfaces.
• This nanotailored immobilization strategy significantly enhances bioreceptor accessibility, leading to improved biomolecular interactions and superior biosensor performance.
• The developed DNA origami-based approach represents a significant advancement towards the next generation of highly sensitive and specific diagnostic sensing devices.