Anion-Exchange Separation of Whey Proteins

초록

In general whey is dilute liquid composed of lactose, a variety of proteins, minerals, vitamins and fat. Whey contains about 6% solids of which 70% or more is lactose and about 0.7% is proteins. Whey protein components are α-Lactalbumin, β-Lactoglobulin, immunoglobulins A, M and G, bovine serum albumin (BSA), lactoferrin and lactoperoxiedase. â-Lactoglobulin is the major whey protein in bovine milk. Bovine whey proteins have potential applications in veterinary medicine, food industrial and as supplements for cell culture media. Three strong anion-exchange membranes (CIM QA, Q100 and HiTrap Q) and reversed-phase high-performance liquid chromatography(15 ㎛　particle with a pore size of 300 Å) were investigated for the separation of the major proteins, which were contained in whey, such as α-Lactalbumin, BSA and β-Lactoglobulin contained. Experiments were performed to determine the optimum mobile phase composition for separating the whey proteins using the standard chemicals of the proteins. For strong anion-exchange membranes, the mobile phase was buffer A (20 mM piperazine-HCl pH 6.4) and buffer B (buffer A + 1 M NaCl) and the linear gradient elution changes of salt concentration were applied. For Reversed-phase high-performance liquid chromatography, the mobil phase were consisted of a linear gradient of the two mobile phases of 0.1% trifluoroacetic acid in water and 0.1% trifluoroacetic acid in acetonitrile. The standard chemicals of the proteins were used to investigate the optimal mobile phase compositions with the three anion-exchange membranes