Achieving high hybridization density at DNA biosensor surfaces using branched spacer and click chemistry

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Diagnostics

Background:

• The COVID-19 pandemic underscored the need for rapid, sensitive, and selective virus detection technologies.
• Surface-based DNA-biosensors offer a promising platform for virus detection, but require precise control over DNA probe immobilization.
• High hybridization density on the sensor surface is critical for achieving a low limit of detection.

Purpose of the Study:

• To develop a novel strategy for immobilizing DNA probes onto borosilicate slides using peptides as anchoring units.
• To create highly sensitive and selective DNA-biosensors for potential application in rapid virus detection.
• To optimize DNA probe density for enhanced biosensor performance.

Main Methods:

• Functionalization of borosilicate slides with DNA probes via peptide anchoring units.
• Utilized copper-catalyzed click chemistry for DNA immobilization, with a copper-free variation also explored.
• Characterization of the resulting biochips to assess hybridization density and probe attachment.

Main Results:

• Successfully immobilized DNA probes onto borosilicate slides using a peptide-based strategy.
• Achieved a high DNA hybridization density of 2.9 pmol per cm² on the functionalized biochips.
• Demonstrated the effectiveness of both copper-catalyzed and copper-free click chemistry approaches for immobilization.

Conclusions:

• Peptide-based immobilization is an effective strategy for achieving high DNA probe density on biosensor surfaces.
• The developed biochips show potential for creating highly sensitive DNA-biosensors for accurate virus detection.
• This approach offers a pathway towards developing faster and more precise diagnostic tools.