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Cation exchange displacement batch chromatography of proteins guided by screening of protein purification parameters
Author(s) -
Kotasińska Marta,
Richter Verena,
Thiemann Joachim,
Schlüter Hartmut
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201200329
Subject(s) - displacement chromatography , chromatography , chemistry , ion chromatography , fast protein liquid chromatography , chromatography column , displacement (psychology) , affinity chromatography , countercurrent chromatography , gel permeation chromatography , column chromatography , high performance liquid chromatography , enzyme , biochemistry , organic chemistry , psychology , psychotherapist , polymer
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.