[Separation and purification of proteins on monolithic anion-exchange columns].
A monolithic anion-exchange column with glycidyl methacrylate as the functional monomer and ethylene dimethacrylate as the cross linker was prepared by a free radical polymerization. The epoxide groups of the column were modified respectively by triethylamine, diethylamine and ethylenediamine that afforded anionic functionalities 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 for the separation of proteins. For chromatographic analysis the effects of mobile phase composition and pH on the separation were investigated. The optimum separation for bovine serum albumin, lysozyme and glutathione was achieved with a gradient elution of mobile phase A (0.01 mol/L Tris-HCl (pH 7.0)) and mobile phase B (mobile phase A + 1.0 mol/L NaCl) with a flow rate of 1.0 mL/min at 25 degrees C. The optimum purification for cellulase enzyme was obtained with a gradient elution of mobile phase A (0.01 mol/L Tris-HCl (pH 7.1)) and mobile phase B (mobile phase A + 1.0 mol/L KBr) with the same flow rate and temperature. The columns exhibited good stability, and cellulase enzyme could be separated and purified quickly on the monolithic anion-exchange column modified by diethylamine.
Zhang
Yihua
Y
College of Pharmacy, Hebei University, Baoding 071002, China.
Yang
Gengliang
G
Zhang
Xiaole
X
Zhao
Jumin
J
Cai
Liping
L
Chen
Yi
Y
D000837:Anion Exchange Resins; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005978:Glutathione / Q000302:isolation & purification; D009113:Muramidase / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D000837:Anion Exchange Resins; D011506:Proteins; D012710:Serum Albumin, Bovine; D009113:Muramidase; D005978:Glutathione