Engineering Two-Dimensional Nanobody-Origami Architectures for Enhanced Antiviral Activity

Area of Science:

• Nanotechnology
• Virology
• Immunology

Background:

• Nanoscale organization of binding ligands is a key strategy for combating viral infections.
• Multivalent nanobodies (Nb) show promise as antiviral agents.
• Controlling the spatial arrangement of Nbs is crucial for enhancing their efficacy.

Purpose of the Study:

• To develop a programmable 2D DNA origami platform for nanoscale control of multivalent Nb spatial patterns.
• To engineer origami-Nb nanoarchitectures with rationally designed Nb patterns for broad-spectrum antiviral applications.
• To evaluate the impact of Nb spatial patterns on viral binding affinity and neutralization potency.

Main Methods:

• Utilized a 2D DNA origami platform for programmable spatial positioning of Nbs.
• Employed site-selective Nb-DNA conjugation for precise Nb attachment.
• Synthesized origami-Nb nanoarchitectures with Nb patterns designed to mimic viral surface protein geometry.
• Tested the efficacy of engineered nanoarchitectures against SARS-CoV-2 and HIV-1.

Main Results:

• Achieved nanoscale control over Nb spatial patterns on a 2D DNA origami platform.
• Demonstrated significantly enhanced viral binding affinity and neutralization potency with optimized Nb configurations.
• For SARS-CoV-2, a triangular Nb pattern achieved an IC50 of 1.52 nM, a 171-fold improvement over monomeric Nbs.
• For HIV-1, an optimal Nb pattern increased neutralization efficiency by 233-fold.

Conclusions:

• The programmable 2D DNA origami platform enables precise nanoscale control of Nb spatial patterns.
• Spatially optimized Nb patterns lead to significantly enhanced viral binding and neutralization.
• This generalizable strategy offers a promising approach for engineering potent antiviral inhibitors targeting viral surface antigens.