title: Sample displacement batch chromatography of proteins.
abstract: In downstream processing large scale chromatography plays an important role. For its development screening experiments followed by pilot plant chromatography are mandatory steps. Here we describe fast, simple, and inexpensive methods for establishing a preparative chromatography for the separation of complex protein mixtures, based on sample displacement batch chromatography. The methods are demonstrated by anion-exchange chromatography of a human plasma protein fraction (Cohn IV-4), including the screening step and scaling up of the chromatography by a factor of 100. The results of the screening experiments and the preparative chromatography are monitored by SDS-PAGE electrophoresis. In summary we provide a protocol which should be easily adaptable for the chromatographic large scale purification of other proteins, in the laboratory as well as in industry for commercial manufacturing. For the latter these protocols cover the initial piloting steps for establishing a sample batch chromatography based on packed columns rather than batch chromatography.
authors.affiliation: Institute of Clinical Chemistry - Mass Spectrometric Proteomics, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Marta
authors.initials: M
authors.lastname: Kotasinska
authors.forename: Verena
authors.initials: V
authors.lastname: Richter
authors.forename: Marcel
authors.initials: M
authors.lastname: Kwiatkowski
authors.forename: Hartmut
authors.initials: H
authors.lastname: Schlüter
mesh_terms: D001798:Blood Proteins / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans
chemical_list: D001798:Blood Proteins