[Cibacron blue F3GA-attached 2 microns non-porous monodisperse silicas for affinity chromatography]

Non-porous monodisperse silica (NPS), 2 microns in diameter, was modified with 3-aminopropyltriethoxysilane for immobilization of Cibacron Blue F3GA (CB), a packing of NPS-ACB for affinity chromatography was obtained. Up to 2 mg of CB could be attached to 1 mL of NPS beads. There was no obvious leakage of dye from NPS-ACB. Oval was not retained by the column, while Lys was specifically adsorbed. The adsorption of Lys varied with pH values and ionic strengths. In addition, alpha-globulin could not be retained by the packing, while beta- and gamma-globulin could be adsorbed on the column. gamma-Globulin was able to be eluted by 20% 1,6-hexanediol and 1 mol/L KCl, while beta-globulin was not able to be eluted by the same eluent. The difference in affinity interaction could be used to separate the three globulins. Furthermore, the column could be used for separation and preparation of Lys from hen egg white. The chromatograms of Lys on non-porous silica diethylamine column (NPS-DEA) showed that retention time of one peak of the crude Lys prepared was in accordance with Lys's, so it could be said that NPS-ACB column can be used for preparation in a small scale.