Synthesis And Characterization Of Nonporous And Rigid Chromatography Matrix

This dissertation aims at synthesizing a rigid, nonporous polymeric bead which can be utilized in fast purification and separation of assorted proteins. In the fabrication of the polymeric bead, Glycidyl methacrylate (GMA) was used as monomer and polyvinylpyrrolidone (PVP) was added as the stabilizer. Moreover, trimethylolpropane trimetacrylate (TRI-TRI) was introduced as the cross linker and polymerization was initiated by 2, 2'-Azobis (isobutyronitrile) (AIBN) at 72℃. In the fabrication of bead, several factors affecting the characteristics of beads, such as volumetric ratio of water to ethanol was investigated in detail to optimize the synthesis. Polymeric bead prepared above was characterized with environmental scanning electric microscopy (ESEM), particle size analyzer (PSA) and high performance liquid chromatography (HPLC). Thus, surface morphography, diameter and mechanic strength of the beads were determined. The results indicated that the average diameter of the beads was 4.9 micrometers with monodispersion plus smooth surface and ideal mechanical strength.Then the beads were modified with sulfonate groups to prepare a kind of cationic exchanger. Two approaches are mentioned in this dissertation: direct reaction with sodium sulfite versus grafting of a polymeric ligand, 2-acylamido-2-methyl-1-propane -sulfonic acid (AMPS). The second reaction was catalyzed by Cerium (IV) and the graft of sulfonate groups on the bead was shown by FTIR.In static adsorption experiment, lysozyme was used as the model protein to test the adsorption capacity of the two beads. Parallel experiments were conducted and demonstrated a significant increase in protein adsorption amount after graft of AMPS. Finally, PGMA-SO3 beads were packed into a stainless column and connected to HPLC. Separation of proteins with different isoelectricity points was carried out later, thus exhibiting the stability and efficiency of the beads. Meanwhile, this'graft to'method could also provide flexible solutions in modification of matrix surface to provide rich chemistries.