Monodisperse thermosensitive new polymer stationary phase chromatography and its application to the separation of biological macromolecules.doc

 PAGE \* MERGEFORMAT 14 Monodisperse thermosensitive new polymer stationary phase chromatography and its application to the separation of biological macromolecules Study: Lung Sun Yanpu GONG Bo-Lin Ouyang Kang [Abstract] 2.5 μm monodisperse polystyrene seeds, ethylene glycol dimethacrylate (EDMA) as crosslinker, toluene and cyclohexanol as porogen, the use of “step seed swelling and polymerization” Preparation of monodisperse microspheres; then potassium persulfate as initiator will be water-soluble temperature sensitive monomer N-isopropyl acrylamide (NIPAM) molecules to the microsphere surface initiated polymerization, particle size prepared 7.0 μm, coefficient of dispersion 0.02 monodisperse cross-linked temperature sensitive chromatographic packing, temperature sensitive monomer NIPAM grafting rate of 5.2%. examine the standard of the packing of protein separation performance, temperature sensitive performance, stability and reproducibility and the dynamic adsorption capacity of retention of proteins. Experimental results show that the chromatographic packing for protein separation performance, temperature sensitive performance, stability and good reproducibility, and the dynamic adsorption capacity of lysozyme was 32.3 mg / g. In the hydrophobic mode, the Not only can fill the baseline separation of 5 standard proteins, and by changing the temperature can effectively be three kinds of overlapping retention time at low temperature, the protein (cytochrome C, β-lactoglobulin and ribonuclease) completely separated. [Keywords:] N-isopropyl acrylamide; monodisperse thermo-sensitive chromatographic stationary phase; protein separation Abstract By using 2.5 μm monodisperse polystyrene as seeds, ethylene glycol dimethyl acrylate (EDMA) as crosslinking agent, toluene and cyclohexanol as porogen, one step of seed swelling polymerization method was applied for the preparation of monodisperse beads. Then the monomer of N isopropylacrylamide ( NTPAM) was po