Preparation of New Polymeric Monodisperse Thermosensitive Chromatographic Stationary Phase and Its Application in Separation of Biopolymers

Using 2.5-μm monodisperse polystyrene as seeds, ethylene glycol dimethylacrylate (EDMA) as crosslinking agent, toluene and cyclohexanol as porogen, one-step of seed swelling polymerization method was used for preparing monodisperse beads. Then, the monomer of N-isopropylacrylamide was polymerized on the surface of monodisperse beads to prepare a 7.0-μm monodisperse thermosensitive chromatographic stationary phase. The stationary phase was prepared by very simple one-step seed swelling polymerization, and the bonding capacity of thermosensitive monomer was controlled easily according to the amount between thermosensitive monomer and crosslinker. The stationary phase was evaluated by determining its separation ability and the effect of temperature on retention of proteins. The dynamic protein loading capacity of the synthesized stationary phase was 32.3 mg g−1. In the hydrophobic interaction chromatographic model, five standard proteins can be separated by the stationary phase, and Cyt-C, β-lact, Rnase-A can be effectively separated by changing the temperature.