A Versatile Virus-Mimetic Engineering Approach for Concurrent Protein Nanocage Surface-Functionalization and Cargo...

Area of Science:

• Biotechnology
• Nanotechnology
• Biochemistry

Background:

• Protein nanocages, such as ferritin, are biocompatible and self-assembling structures with biomedical potential.
• Existing engineering methods for nanocage-based drug delivery and vaccine development are limited in scope and efficiency.

Purpose of the Study:

• To develop a versatile and straightforward engineering approach for creating protein nanocages for drug delivery and vaccine applications.
• To mimic the HIV-1 Gag polyprotein precursor to design novel ferritin-like nanocages.

Main Methods:

• Designed and synthesized various PREcursors of nanoCages (PREC) by linking ferritin subunits with a protease cleavage site.
• Utilized protease cleavage to generate protease-induced nanocages (PINCs) with potential for N-terminal protein addition.
• Demonstrated concurrent surface decoration and drug encapsulation (hydrophilic/hydrophobic) within PINCs.

Main Results:

• PINC formation enabled simultaneous surface functionalization and drug encapsulation, achieving up to fourfold higher drug loading than other methods.
• The developed PINCs/Drug complex exhibited stability and efficacy in killing cancer cells.
• Provided insights into PREC design rules and the mechanism of PINC formation.

Conclusions:

• The novel engineering approach using PREC precursors provides a versatile platform for creating protease-induced nanocages (PINCs).
• PINCs offer enhanced capabilities for drug delivery and multifunctional therapeutic development, including mosaic vaccines.
• This work facilitates the advancement of nanocage applications in nanomedicine.