Adsorption and elution of bovine gamma-globulin using an affinity membrane containing hydrophobic amino acids as ligands.
A hollow-fibre affinity membrane containing hydrophobic amino acids as ligands was prepared by the radiation-induced grafting of glycidyl methacrylate onto a porous polyethylene hollow fibre and subsequent phenylalanine (Phe) or tryptophan (Trp). The densities of the Phe and Trp ligand of the resulting affinity membrane were 0.4 and 0.4 mol/kg, respectively. The Trp-containing affinity membrane exhibited a higher amount of adsorbed bovine gamma-globulin (BGG) than the Phe-containing membrane. To evaluate the adsorption behaviour of the membrane, the BGG-containing buffer solution was permeated from the inside to the outside of the Trp-containing hollow-fibre affinity membrane through the ligand-immobilized pores. The breakthrough curves as a function of effluent volume coincided irrespective of the flow-rate, i.e. the residence time (55-220 s) of the solution across the membrane (thickness 0.83 mm), as a result of negligible mass transfer resistance. A series of chromatographic procedures, (adsorption-washing-elution) was repeated twice and a satisfactory quantitative elution was attained. The reproducible profile of the flux and the protein concentration assured a quantitative cycle of chromatography using the affinity membrane containing Trp as a ligand.
Kim
M
M
Department of Chemical Engineering, Faculty of Engineering, University of Tokyo, Japan.
Saito
K
K
Furusaki
S
S
Sato
T
T
Sugo
T
T
Ishigaki
I
I
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D008024:Ligands; D010649:Phenylalanine; D014364:Tryptophan; D005719:gamma-Globulins / Q000032:analysis*
D008024:Ligands; D005719:gamma-Globulins; D010649:Phenylalanine; D014364:Tryptophan