Precise modulation of protein refolding by rationally designed covalent organic frameworks

Area of Science:

• Materials Science
• Biotechnology
• Chemical Engineering

Background:

• Precisely controlling protein conformation is crucial for biomanufacturing and biomedicine.
• Current methods for protein refolding face significant challenges in efficiency and specificity.

Purpose of the Study:

• To develop a novel covalent organic framework (COF)-directed strategy for efficient and customizable protein refolding.
• To investigate the mechanism of COF-mediated protein refolding and its applicability across various proteins.

Main Methods:

• Utilized rationally designed covalent organic frameworks (COFs) with tailored pore structures and microenvironments.
• Employed a one-step COF treatment in aqueous or buffer solutions for denatured protein refolding.
• Investigated refolding mechanisms through analysis of pore size and microenvironmental factors (hydrophobicity, π-π conjugation, hydrogen bonding).

Main Results:

• Achieved efficient protein refolding for diverse proteins including lysozyme, glucose oxidase, trypsin, nattokinase, and papain.
• Demonstrated high generality across various proteins and COF materials.
• Developed solid-phase columns for continuous protein recovery with ~100% refolding yield and 30-cycle recyclability.

Conclusions:

• The COF platform provides a highly efficient and customizable method for precise protein refolding.
• This strategy enables integrated processes for extracting and refolding denatured proteins, such as from inclusion bodies.
• Opens new avenues for advanced protein manufacturing and biomedical applications.