Sample displacement batch chromatography of proteins
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Sample displacement batch chromatography of proteins. / Kotasinska, Marta; Richter, Verena; Kwiatkowski, Marcel; Schlüter, Hartmut.
In: Methods Mol Biol, Vol. 1129, 01.01.2014, p. 325-38.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Sample displacement batch chromatography of proteins
AU - Kotasinska, Marta
AU - Richter, Verena
AU - Kwiatkowski, Marcel
AU - Schlüter, Hartmut
PY - 2014/1/1
Y1 - 2014/1/1
N2 - 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.
AB - 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.
U2 - 10.1007/978-1-62703-977-2_24
DO - 10.1007/978-1-62703-977-2_24
M3 - SCORING: Journal article
C2 - 24648085
VL - 1129
SP - 325
EP - 338
JO - Methods Mol Biol
JF - Methods Mol Biol
SN - 1064-3745
ER -