title: Capture of proteins from mammalian cells in pilot scale using different STREAMLINE adsorbents.
abstract: This presentation compares three different expanded bed matrices. STREAMLINE rProtein A, STREAMLINE SP-XL and STREAMLINE Chelating were monitored in respect to their ability to clarify the broth, to concentrate and to purify the distinct target protein. The capture of a mouse IgG1 and a recombinant prothrombin (PT) was carried out in pilot scale using a 100-l bioreactor and STREAMLINE 100 and 200 columns, respectively. The robustness of the process was also estimated monitoring the expansion behaviour and the cell and debris concentrations during the load and in the eluat. In all cases the capture of the target proteins was comparable to conventional chromatographic systems. The purification success was mainly dependent on the selectivity of the ligand used. The affinity process resulted in a highly purified product. The ion exchanger and chelating material mainly concentrated the product. In all three cases 100 l of cell broth were successfully processed in one run. The robustness of the ion exchanger process was poor, because of strong cell matrix interaction. However, for the chelating and especially for the affinity matrix a highly reproducible process was obtained.
authors.affiliation: Cell Culture Technology, Faculty of Technology, Institute of Cell Culture Technology, University of Bielefeld, Germany. dlu@zellkult.techfak.uni-bielefeld.de
authors.forename: D
authors.initials: D
authors.lastname: Lütkemeyer
authors.forename: N
authors.initials: N
authors.lastname: Ameskamp
authors.forename: C
authors.initials: C
authors.lastname: Priesner
authors.forename: E M
authors.initials: EM
authors.lastname: Bartsch
authors.forename: J
authors.initials: J
authors.lastname: Lehmann
mesh_terms: D000327:Adsorption; D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D051379:Mice; D010865:Pilot Projects; D011516:Prothrombin / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D007074:Immunoglobulin G; D011994:Recombinant Proteins; D011516:Prothrombin