Sample Displacement Batch Chromatography of Proteins.

Laura Heikaus, Siti Hidayah, Manasia Gaikwad, Marta Kotasinska, Verena Richter, Marcel Kwiatkowski, Hartmut Schlüter

Methods in molecular biology (Clifton, N.J.) 2021

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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 upscaling of the chromatography by a factor of one hundred. 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 the manufacturing of biopharmaceuticals. These protocols cover the initial piloting steps for establishing a large-scale sample batch chromatography. The results from the piloting steps may also be applied for packed columns for performing simulated-moving-bed (SMB) chromatography rather than batch chromatography.

Citation

Laura Heikaus, Siti Hidayah, Manasia Gaikwad, Marta Kotasinska, Verena Richter, Marcel Kwiatkowski, Hartmut Schlüter. Sample Displacement Batch Chromatography of Proteins. Methods in molecular biology (Clifton, N.J.). 2021;2178:285-299