Biotechnology Progress, Vol.33, No.4, 1104-1115, 2017
Influence of Mixed Electrolytes on the Adsorption of Lysozyme, PEG, and PEGylated Lysozyme on a Hydrophobic Interaction Chromatography Resin
In a recent work (Werner A and Hasse H in J Chromatogr A 2013; 1315: 135) the influence of mixed electrolytes on the adsorption of the macromolecules lysozyme, PEG and di-PEGylated lysozyme on a hydrophobic resin has been studied, but only at one overall ionic strength (3000 mM). The present work, therefore, extends these studies to other ionic strengths (2400 and 2700 mM), and explores the application of a model to predict the entire data set. The adsorbent is Toyopearl PPG-600M. The solvent is a 25 mM aqueous sodium phosphate buffer at pH 7.0. The studied salts are sodium chloride, sodium sulfate, ammonium chloride and ammonium sulfate. Pure salts as well as binary and ternary mixtures of these salts with varying ratios of the amounts of the salts are studied at 25 degrees C. The loading of the adsorbent increases with increasing salt concentration for all macromolecules. Synergetic effects of the mixed electrolytes are observed. (C) 2017 American Institute of Chemical Engineers
Keywords:hydrophobic interaction chromatography (HIC);mixed electrolytes;PEG;modeling;lysozyme;ionic strength