Characterization and application of strong ion-exchange membrane adsorbers as stationary phases in high-performance liquid chromatography of proteins.
Filtration membranes carrying strong cation- or anion-exchange groups on their surface were evaluated for their potential as membrane adsorber stationary phases in the high-performance liquid chromatography of proteins. The membranes are commercially available and can be obtained inserted into ready-to-use filter holders. Owing to their thinness (170-190 microns), the pressure drop of the membranes is extremely low. Flow-rates of up to 65 ml min-1 per unit became thus possible. The low pressure drop of a single membrane layer also permitted an effortless scaling up, as a stack of several membranes or filter units could be used, if necessary. Sample distribution, protein binding capacity, elution conditions, separation efficiency and recovery were investigated as a function of the flow-rate. The time required for the separation of certain protein mixtures could be reduced to less than 1 min. Appropriate conditions were defined for the separation of human serum and for the isolation of subtilisin Carlsberg and beta-galactosidase from cell culture supernatants.
Reif
O W
OW
Institut für Technische Chemie, Universität Hannover, Germany.
Freitag
R
R
D000837:Anion Exchange Resins; D002411:Cation Exchange Resins; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006801:Humans; D008567:Membranes, Artificial; D011506:Proteins / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet
D000837:Anion Exchange Resins; D002411:Cation Exchange Resins; D008567:Membranes, Artificial; D011506:Proteins