Hydrophobic Interaction Chromatography
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Hydrophobic Interaction Chromatography.
Most proteins and large polypeptides have hydrophobic regions at their surface. These hydrophobic "patches" are due to the presence of the side chains of hydrophobic or nonpolar amino acids such as phenylalanine, tryptophan, alanine, and methionine. These surface hydrophobic regions are interspersed between more hydrophilic or polar regions and the number, size, and distribution of them is a sp...
متن کاملCalmodulin interacts with cyclic nucleotide phosphodiesterase and calcineurin by binding to a metal ion-independent hydrophobic region on these proteins.
Hydrophobic interaction chromatography is employed to determine if calmodulin might associate with its target enzymes such as cyclic nucleotide phosphodiesterase and calcineurin through its Ca2+-induced hydrophobic binding region. The majority of protein in a bovine brain extract that binds to a calmodulin-Sepharose affinity column also is observed to bind in a metal ion-independent manner to p...
متن کاملHydrophobic Interaction Chromatography: Fundamentals and Applications in Biomedical Engineering
Hydrophobic interaction chromatography (HIC) a powerful technique used for separation and purification of biomolecules. It was described for the first time by Shepard & Tiselius (1949), using the term “salting-out chromatography”. Later, Shaltiel & Er-el (1973) introduced the term “hydrophobic chromatography”. Finally, Hjerten (1973) described this technique as “hydrophobic interaction chromato...
متن کاملModeling of adsorption in hydrophobic interaction chromatography systems using a preferential interaction quadratic isotherm.
A preferential interaction quadratic isotherm model for hydrophobic interaction chromatographic systems is presented in this paper. In this isotherm, the nonlinear effect of salt on the capacity factor is described using the preferential interaction model developed by Perkins et al. [J. Chromatogr. A, 766 (1997) 1]. This is then coupled with a quadratic nonlinear isotherm to describe nonlinear ...
متن کاملA Simple Method for the Estimation of Protein Retention in Hydrophobic Interaction Chromatography Under Different Operation Conditions
Protein behavior in Hydrophobic Interaction Chromatography using different chromatographic conditions was investigated. A linear correlation was found between protein retention time on different matrixes and different initial elution salt concentrations. Mathematical correlations between retention times under different chromatographic conditions were obtained and validated, which can be used in...
متن کاملPurification of Plasmid (pVaxLacZ) by Hydrophobic Interaction Chromatography
This paper describes a method for the plasmid DNA purification, which includes an ammonium sulphate precipitation, followed by hydrophobic interaction chromatography (HIC) using Phenyl Sepharose 6 Fast Flow (low sub). The use of HIC took advantage of the more hydrophobic character of single stranded nucleic acid impurities as compared with double-stranded plasmid DNA.
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