Preparation of iron porphyrin-based polymer monolithic column via atom transfer radical polymerization for separation of proteins and small molecules
A novel iron porphyrin-based polymer monolithic column (50 × 4.6 mm i.d.) was prepared via atom transfer radical polymerization for separation of proteins from complex bio-matrix, taking advantages of specific absorption to protein of iron porphyrin and skeleton porous structure with high surface area as 270.3 m2 g-1 of the monolith. In the preparation processes, iron porphyrin and glycidyl methacrylate were used as co-monomers; ethylene dimethacrylate was used as cross linking agent, polyethylene glycol 200 and 1,4-butanediol were used as co-porogens, N, N-dimethylformamide was used as solvent; CCl4 and FeCl2 were used as initiator and catalyst, respectively. Characteristics of the optimized monolithic columns were investigated by elementary analyzer, scanning electron microscope, thermogravimetric analysis, Brunauer-Emmett-Teller nitrogen adsorption-desorption, and mercury intrusion porosimeter, respectively. The results showed that the iron porphyrin-based monolithic column had improved skeleton porous structure with micro-, meso- and macropores. The monolith was successfully used to separate both protein with high resolution and small molecules with highest column efficiency of 29 000 plates per meter. All the results confirmed that the present method would be scientific valuable for both classifiable protein analysis and small molecular separation.