Adsorption Phenomena in Hydrophobic Interaction Chromatography

Autor: Belinda F. Roettger, Fred E. Regnier, Michael R. Ladisch, Julia A. Myers
Rok vydání: 1989
Předmět:
Zdroj: Biotechnology Progress. 5:79-88
ISSN: 1520-6033
8756-7938
DOI: 10.1002/btpr.5420050304
Popis: Hydrophobic interaction chromatography (HIC) separates proteins on the basis of surface hydrophobicity while generally retaining the activity of proteins. Aqueous mobile phases with high salt concentrations are often used to adsorb the proteins onto a mildly hydrophobic support. HIC protein adsorption is modeled using the thermodynamic theory of linked functions (i.e., Wyman's linkage theory). This approach correlates adsorption to preferential interactions of the salts with supports and proteins. Two different methods were employed to study the salt interactions with the support: chromatography retention studies and densimetric techniques. Results of the salt chromatography trials indicate that chaotropic salts such as NaSCN and NaI have positive preferential interaction parameters that decrease with increasing concentration. Densimetric measurements show that the preferential interaction parameters of polar kosmotropic salts, such as ammonium sulfate are negative and become more negative with increasing concentration. The correlation of HIC adsorption to preferential interaction parameters was experimentally verified by retention studies of lysozyme in aqueous chaotropic and kosmotropic mobile phases, which also show a linear relation between the protein's capacity factor and the lyotropic series.
Databáze: OpenAIRE
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