Boolean Logic-Based Controlled Release of Bioactive Proteins with Diversified Inputs.

Area of Science:

• Biomaterials Science
• Molecular Engineering
• Synthetic Biology

Background:

• Stimuli-responsive biomaterials are crucial for controlled therapeutic delivery, tissue engineering, and biosensing.
• Previous work utilized autonomous compilation for protein release from biomaterials based on Boolean logic, but was limited to simple outputs and inputs.
• Expanding this framework is essential for broader applications in biomedicine.

Purpose of the Study:

• To extend the capabilities of logic-based protein release from biomaterials.
• To demonstrate the assembly of all seven unique logical operations using a 3-input operator set.
• To enable the release of diverse bioactive proteins in response to specific stimuli.

Main Methods:

• Assembled all 7 unique logical operations (YES/OR/AND) for protein release control.
• Utilized three orthogonal protease actuators to trigger conditional release of protein cargos.
• Incorporated a photocleavable protein motif to enable visible light as an additional release input.

Main Results:

• Successfully demonstrated the programmable release of diverse bioactive proteins, including growth factors, enzymes, nanobodies, cytokines, and fluorescent proteins.
• Achieved logic-based release from material anchors in response to precise combinations of protease inputs, maintaining protein bioactivity.
• Showcased visible light as a controllable input for logic-based protein release.

Conclusions:

• The developed framework significantly expands the applicability of logic-based protein release from biomaterials.
• This approach enables precise control over the release of various functional biomacromolecules.
• The findings pave the way for advanced applications in targeted therapeutics, diagnostics, and synthetic biology.