Simplification of affinity macroporous monolith microfluidic column synthesis and its ability for protein separation

Area of Science:

• Materials Science
• Chromatography
• Biochemistry

Background:

• Developing efficient chromatography resins is crucial for protein purification.
• Simultaneous polymerization and ligand immobilization offers a facile route to advanced materials.

Purpose of the Study:

• To design a generic multi-component approach for simultaneous in situ polymerization and ligand immobilization.
• To develop a facile method for creating affinity porous polymer-based chromatography resin.

Main Methods:

• Utilized a regioselective ring-opening reaction between epoxy groups and amine functional groups under aqueous conditions (pH 9.7).
• Demonstrated proof-of-concept using iminodiacetic acid (IDA) and allyl glycidyl ether (AGE) with HEMA, DATD, and PDA monomers via thermal initiation at 65°C for 4 hours.
• Confirmed polymerization via visual observation and SEM, and ligand immobilization via FT-IR analysis.

Main Results:

• Successful polymerization and ligand immobilization were achieved.
• The iminodiacetic acid (IDA)-functionalized monolith chelated copper (Cu II) ions.
• The resulting affinity column exhibited an IgG adsorption capacity of 27.8 mg/g monolith.
• Efficient capture of high-abundance proteins (IgG, transferrin, albumin) from human plasma was demonstrated.

Conclusions:

• The developed generic approach enables facile simultaneous in situ polymerization and ligand immobilization for creating affinity chromatography resins.
• The metal-chelating IDA-functionalized porous polymer demonstrates significant protein adsorption capacity and efficient capture of target proteins from complex biological samples like human plasma.