Engineering and functionalization of large circular tandem repeat protein nanoparticles.

Area of Science:

• Biotechnology and protein engineering
• Nanotechnology and materials science
• Immunology and cell biology

Background:

• Protein engineering allows for the creation of diverse molecular scaffolds.
• Symmetric protein nanoparticles offer enhanced avidity, stability, and solution behavior.
• Displaying multiple protein domains on nanoparticles can improve functional properties.

Purpose of the Study:

• To design and characterize a computationally designed circular tandem repeat protein (cTRP) nanoparticle.
• To demonstrate the self-assembly and versatility of cTRP nanoparticles for displaying functional domains.
• To explore the application of cTRP nanoparticles in high-avidity molecular interactions, such as T-cell expansion.

Main Methods:

• Computational protein design of a circular tandem repeat protein (cTRP).
• Characterization of self-assembly properties of cTRP subunits into nanoparticles.
• Demonstration of various cargo attachment strategies for functional domain display.
• Evaluation of cTRP nanoparticle performance in T-cell culture and expansion.

Main Results:

• Successful creation and characterization of computationally designed cTRP nanoparticles.
• Demonstrated self-assembly of cTRP nanoparticles from individual subunits.
• Confirmed production of cTRP nanoparticles from both prokaryotic and human expression systems.
• Showcased the utility of cTRP nanoparticles in applications requiring high-avidity cell surface interactions.

Conclusions:

• cTRP nanoparticles represent a versatile platform for displaying multiple protein domains with enhanced avidity.
• These nanoparticles can be produced using various expression systems and attachment strategies.
• cTRP nanoparticles show significant potential for applications in cell-based therapies and research, including T-cell expansion.