Separation And Purification Of Proteins And Application Of Drugs In Human Body On Ion-Exchange Monolithic Column

Ion-exchange monolithic columns, which were developed greatly in recent years, have been widely used in separation and purification of protein, peptide, nucleic acid, polyribonucleotide, and polysaccharide. They have many excellent characteristics, such as 1) Open-ended structure, allowing macromolecules to distribute and pass through quickly, by which rapid separation and preparation could be obtained; 2) Porosity, high superficial area and exchange capacity; 3) Hydrophilicity, which would not result in the inactivation of protein and enzyme, because of their weak adsorption to bio-macromolecules, and high active recovery. Furthermore, they can also be used as the material of solid phase extraction (SPE) in the preconcentration of trace components in blood before further analysis. Compared with conventional enrichment technique, it is safe, convenient, effective and it will make automatic on-line analysis possible without overfull consumption of organic solvent.In the paper, monolithic column was prepared by "in-situ" polymerization, and ion-exchange groups were bonded on the column by "on-line" modification. It was investigated that separation and purification of cellulase enzyme on the anion-exchange monolithic column and simultaneously deproteinization and enrichment of drugs on the weak cation-exchange monolithic column. There are three chapters in this thesis.Chapter one: A review mainly concerned with the recent progress in the stationary phase of ion-exchange chromatography (IEC), including inorganic, organic and composite matrices, and the applications of IEC in biochemistry.Chapter two: An anion-exchange monolithic column was prepared by "in-situ" polymerization. The epoxide groups of the column were modified respectively by triethylamine, diethylamine and ethylenediamine that afforded anionic function groups which were required for the anion-exchange chromatographic mode. The properties of the monolithic columns were investigated and the columns were successfully used as stationary phases of high performance liquid chromatography (HPLC) for the separation of proteins. The columns exhibited good stability, and cellulase enzyme could be separated and purified quickly on the anion-exchange monolithic column...