Ion selectivity in the high-performance cation-exchange chromatography of proteins

Advances in native high‐performance liquid chromatography and intact mass spectrometry for the characterization of biopharmaceutical products

“…and Regnier et al. . Not all charge variants are generally resolved using a salt gradient in IEC mode, especially the acidic variants .…”

“…[91,92]. The effects of eluent salts on the resolution of protein separation has been described by Gooding et al and Regnier et al [93,94]. Not all charge variants are generally resolved using a salt gradient in IEC mode, especially the acidic variants [95].…”

Charged fusions for enhanced protein purification and immobilization by ion-exchange

“…In principle, the unique electrostatic interactions of each protein with the stationary phase forms the basis for separation. However, the amphoteric nature and three-dimensional structure of the proteins make their interaction with the ionexchange surface very complex (Geng and Regnier, 1985;Gooding and Schmuck, 1984;Regnier, 1987). The ion-exchanger (Drager and Regnier, 1987;Wu and Walters, 1992) and the mobile phase Kopaciewicz et al, 1985;Rounds and Regnier, 1984) can also affect the protein retention and subsequent elution.…”

“…suggested that both poor resolution and low recovery can be improved by changing the type of displacing salt. Several studies devoted to studying the effects of different displacing ions on protein retention are available (Gooding and Schmuck, 1984;Rounds and Regnier, 1984). Unfortunately, due to the heterogeneity of protein (both structure and ionic content), the relation between selectivity and mobile phase remains empirical.…”