Encapsidic production and isolation of degradation-prone polypeptides

Area of Science:

• Biotechnology
• Biochemistry
• Molecular Biology

Background:

• Degradation of biomolecular therapies during production and delivery is a major obstacle.
• Current methods struggle to protect labile therapeutic molecules effectively.

Purpose of the Study:

• To develop a robust system for the microbial production and isolation of therapeutic biomolecules.
• To protect fragile biomolecules from degradation throughout the production and delivery pipeline.

Main Methods:

• Utilizing engineered protein cages derived from lumazine synthase.
• Genetically fusing therapeutic polypeptides to cage component protomers for encapsulation.
• Employing sequence-specific protease cleavage for controlled cage opening and cargo isolation.
• Implementing modular patchwork assembly to regulate cage formation and prevent aggregation.

Main Results:

• Demonstrated efficient encapsulation and protection of target polypeptides within protein cages.
• Achieved controlled release of encapsulated cargo via protease-mediated cage opening.
• Successfully produced six distinct, intrinsically disordered polypeptides with therapeutic potential.
• Prevented guest overloading and formation of insoluble aggregates through modular assembly.

Conclusions:

• The protein cage "encapsidic" approach offers a versatile strategy for producing and delivering biomolecular therapies.
• This method significantly enhances the stability and accessibility of labile therapeutic agents.
• The system shows broad applicability for various therapeutic polypeptides, addressing critical production bottlenecks.