Chromatography of proteins on hydrophobic interaction and ion-exchange chromatographic matrices: mobile phase contributions to selectivity

Activation of Alpha Chymotrypsin by Three Phase Partitioning Is Accompanied by Aggregation

“…The aggregate (obtained with 55% saturation of ammonium sulphate and dialyzed against distilled water) was indeed partially dissociated in the presence of ≥10% (v/v) concentration of ethylene glycol (Fig. 3) which is known to decrease hydrophobic interactions [30]. Hence, TPP treatment led to alpha chymotrypsin becoming more prone to aggregation and this aggregation was mediated by hydrophobic interactions.…”

High-throughput isotherm determination and thermodynamic modeling of protein adsorption on mixed mode adsorbents

“…Desorption is induced by charge repulsion, accomplished by changing the pH of the mobile phase across the pI of the protein [7,8] or across the pK a of the ligand in the case of hydrophobic charge induction chromatography (HCIC) [9]. The main advantage of mixed mode resins is the wider range of selectivity that may be achieved in a single column compared to conventional ion exchange chromatography (IEX) or hydrophobic interaction chromatography (HIC) [10]. Hence, one mixed mode chromatographic column can potentially replace two orthogonal chromatographic steps in a protein purification process, with clear cost benefits.…”

High‐Performance Liquid Chromatography of Biological Macromolecules

“…The same group has also investigated the combined use of salts and solvents to affect retention in HIC and IEC. 260 Hearn et al studied the effect of different displacing salts on retention and band broadening of proteins in isocratic anion chromatography. 600 The retention order was found to correlate well with decreasing hydrated ionic radii of the eluent anions and the retention was also affected by the type of cation, an effect attributed to preferential interactions with the protein solute.…”