Preparation of a novel weak cation exchange/hydrophobic interaction chromatography dual-function polymer-based stationary phase for protein separation using “thiol–ene click chemistry”

摘要: A novel dual-function mixed-mode stationary phase based on poly(glycidyl methacrylate-co-ethylene dimethacrylate) microspheres was synthesized by thiol–ene click chemistry and characterized infrared spectroscopy elemental analysis. The new system displays both hydrophobic interaction chromatography (HIC) character in a high salt concentration mobile phase, weak cation exchange (WCX) low phase. It can be used to separate proteins ion-exchange (IEC) mode HIC mode. resolution selectivity of the were evaluated IEC using protein standards. In comparison with conventional WCX columns, results satisfactory acceptable. Protein mass bioactivity recoveries more than 96 % achieved this column. indicate that column many cases replace use two individual columns. addition, effects separation different ligand structures pH also investigated detail. show electrostatic must match hydrophobicity ligand, which is an important factor prepare On basis column, two-dimensional liquid technology single developed study, renature purify recombinant human interferon-γ from inclusion bodies. recovery, purity, specific obtained for purified 87.2 %, 92.4 %, 2.8 × 107 IU/mg, respectively, mode, 83.4 %, 95.2 %, 4.3 × 107 IU/mg, prepared study may aid development intensive proteomic studies, it find applications drug production since save costs simplify biotechnological processes.