Highly Efficient Chiral Separation Based on Alkali-proof Protein Immobilization by Covalent Organic Frameworks

Area of Science:

• Chromatography
• Biochemistry
• Materials Science

Background:

• Chiral separation is crucial for pharmaceuticals and industry.
• Traditional chiral stationary phases (CSPs) lack stability in alkaline environments.

Purpose of the Study:

• To develop ultrastable protein-based CSPs for alkaline conditions.
• To investigate the enantioseparation capabilities of alcalase-based CSPs for acidic compounds.

Main Methods:

• Theoretical simulation of alcalase conformation.
• In situ immobilization of alcalase onto a covalent organic framework (COF) platform.
• High-performance liquid chromatography (HPLC) analysis.

Main Results:

• Developed stable, protein-based CSPs using alcalase and a COF platform.
• Achieved baseline separation (Rs≥1.50) for acidic drugs like RS-flurbiprofen and RS-tenofovir.
• Demonstrated enhanced separation with increased COF hydrophilicity and pore size.
• Exhibited exceptional durability over >2,400 runs.

Conclusions:

• Alcalase-based CSPs provide a stable and effective platform for enantioseparation under alkaline conditions.
• This approach offers superior performance compared to commercial chiral columns.
• The study highlights the potential for protein-based CSPs in separating acidic drugs.