Adenovirus Fibers as Ultra-Stable Vehicles for Intracellular Nanoparticle and Protein Delivery

Area of Science:

• Biotechnology
• Nanomedicine
• Protein Engineering

Background:

• Protein-based carriers offer potential for intracellular therapeutic delivery.
• Adenovirus fiber proteins provide a robust structural framework for nanocarrier development.

Purpose of the Study:

• To engineer adenovirus fiber protein constructs as nanorods for protein and nanoparticle delivery.
• To evaluate the stability, cellular uptake, and cytotoxicity of these novel nanocarriers.

Main Methods:

• Constructed adenovirus fiber protein segments (residues 61-392) with a fibritin foldon trimerization motif.
• Incorporated an N-terminal His-tag and a biotinylation sequence for cargo attachment.
• Assessed nanorod stability, cellular internalization, and cargo delivery in NIH3T3 and HeLa cells.

Main Results:

• Engineered constructs formed thermally and protease-stable fibrous nanorods.
• Nanorods were non-cytotoxic and efficiently internalized by cells.
• Successfully delivered biotinylated gold nanoparticles and enzymatically active proteins (alkaline phosphatase, horseradish peroxidase) into cells via biotin-streptavidin interaction.

Conclusions:

• Adenovirus fiber-based nanorods represent a viable and versatile platform for intracellular delivery of diverse cargoes.
• This proof-of-principle study paves the way for developing multifunctional, modular nanorod systems for targeted therapeutics.