Preparation Of Agarose Chromatograph Media With Controlled Ligand Density Through Direct Coupling Reaction

Hydrophobic interaction chromatography (HIC) is widely used in protein separation and purification. Butyl-agarose, phenyl-agarose and octyl-agarose are the most important HIC media. In this dissertation, the three chromatography media were prepared by direct coupling method and characterization of the three chromatography media were investigated.The three chromatography media were prepared by direct coupling method coupling. In direct coupling reaction, the operation condition, including reaction time, temperature, the amount of boron trifluoride were investigated. The preparative conditions were optimized by orthogonal design of the experiments. The optimum conditions for butyl-agarose were found to be 0.02 mL BF₃·Et₂O/g-gel, reaction time 45 min and reaction temperature 45 ℃. The ligand density could be controlled by the amount of BGE added to the reaction mixture. The optimum conditions for Phenyl-agarose were found to be 0.0lmL BF₃·Et₂O/g-gel, reaction time 30 min and reaction temperature 45℃. The ligand density could be controlled...