Preparation Of Poly GMA Monolithic Columns And Their Application To The Separation Of IL-18

In contrast to traditional packed column, monolithic column has the advantage of easy preparation, excellent permeability, low backpressure at chromatographic runs and large capacity for samples. This kind of column has been applied in many fields.This paper consists of the following parts: preparation of monolithic polymer column, investigation of polymerization conditions and its application to bio-molecules.First, according to the.literatures, a continuous ion-exchange chromatographic column with glycidyl methacrylate as the functional monomer and ethylene dimethacrylate as the cross linker was prepared by a free radical polymerization and applied to the separation of EL-18. After the factors influencing the separation were investigated, an optimum condition for separating IL-18 from its related proteins was obtained.Second, the epoxide groups of the monolithic column were modified respectively by their reaction with triethylamine, diethylamine and ethylenediamine that afford anionic functionalities required for the anion-exchange chromatography of proteins. After the chromatographic capability was investigated, the separation of snailase and its related proteins was abtained with the monolithic column modified by ethylene diamine. Some anions were separated with the monolithic column modified by diethylamine.Third, after the epoxide groups of the monolithic column were modified by its reaction with ethylene diamine, it was modified by its reaction with chloroacetic acid again. Thus a cation-exchange chromatographic column was obtained. The papain was separated from its related proteins with this column; The epoxide groups of the monolithic column were modified in turn with ethylene diamine, chloroacetic acid and coppersulfate, an iminodiacetic acid-bound molded monolithic rod was prepared, and then the separation of superoxide dismutas from its related proteins was obtained with this column; When the matrix of the monolithic column was prepared, it was modified by p-aminoazobenzene to bear affinity properties, and then lysozyme was separated from its related proteins with this column.