Cation exchange displacement batch chromatography of proteins guided by screening of protein purification parameters.
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Cation exchange displacement batch chromatography of proteins guided by screening of protein purification parameters. / Kotasinska, Marta; Richter, Verena; Thiemann, Joachim; Schlüter, Hartmut.
In: J SEP SCI, Vol. 35, No. 22, 22, 2012, p. 3170-3176.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Cation exchange displacement batch chromatography of proteins guided by screening of protein purification parameters.
AU - Kotasinska, Marta
AU - Richter, Verena
AU - Thiemann, Joachim
AU - Schlüter, Hartmut
PY - 2012
Y1 - 2012
N2 - Displacement chromatography has been shown to be an effective alternative for protein purification. We investigated in this study sample displacement chromatography, which does not require a displacer molecule. Furthermore, we performed a screening for determination of parameters for an optimal sample displacement chromatography. We screened the affinities of cytochrome C, lysozyme, myoglobin, and ribonuclease A toward a cation exchange material as a function of different pH values and to presence of different concentrations of sodium chloride in the sample application buffer. Sample displacement chromatography in batch chromatography mode for the separation of the protein mixture was studied with a sample application buffer with a pH of 5 and 7. As predicted by the screening experiments, sample displacement chromatography was most effective at pH 7 since this pH guaranteed the largest differences of the affinities of the four proteins toward the stationary phase. In summary, we describe here sample displacement chromatography in the batch chromatography mode for the separation of proteins, which is a simple and fast alternative to conventional displacement chromatography. Systematic screening of chromatographic parameters prior to sample displacement chromatography promises a successful separation of a target protein.
AB - Displacement chromatography has been shown to be an effective alternative for protein purification. We investigated in this study sample displacement chromatography, which does not require a displacer molecule. Furthermore, we performed a screening for determination of parameters for an optimal sample displacement chromatography. We screened the affinities of cytochrome C, lysozyme, myoglobin, and ribonuclease A toward a cation exchange material as a function of different pH values and to presence of different concentrations of sodium chloride in the sample application buffer. Sample displacement chromatography in batch chromatography mode for the separation of the protein mixture was studied with a sample application buffer with a pH of 5 and 7. As predicted by the screening experiments, sample displacement chromatography was most effective at pH 7 since this pH guaranteed the largest differences of the affinities of the four proteins toward the stationary phase. In summary, we describe here sample displacement chromatography in the batch chromatography mode for the separation of proteins, which is a simple and fast alternative to conventional displacement chromatography. Systematic screening of chromatographic parameters prior to sample displacement chromatography promises a successful separation of a target protein.
KW - Hydrogen-Ion Concentration
KW - Adsorption
KW - Cation Exchange Resins/chemistry
KW - Chromatography, Ion Exchange/instrumentation/methods
KW - Proteins/chemistry/isolation & purification
KW - Hydrogen-Ion Concentration
KW - Adsorption
KW - Cation Exchange Resins/chemistry
KW - Chromatography, Ion Exchange/instrumentation/methods
KW - Proteins/chemistry/isolation & purification
M3 - SCORING: Journal article
VL - 35
SP - 3170
EP - 3176
JO - J SEP SCI
JF - J SEP SCI
SN - 1615-9306
IS - 22
M1 - 22
ER -
