Hydrophobic interaction chromatography of proteins.

摘要: Publisher Summary Hydrophobic interaction chromatography (HIC) is a useful tool for purifying proteins, with maintenance of the biological activity. In this method, hydrophobic ligands (e.g., n-alkyl or phenyl groups) are chemically bound to matrices (polymer silica gels) and protein components, interact these ligands, through application high concentration an anti-chaotropic salt [e.g., (NH4)2S04)]. This diminished, by decreasing concentration, leading elution. chapter, general guidelines, concerning separation conditions in HIC, discussed variety examples Escherichia coli-expressed glycosylated membrane antibodies). Protein structure conformation play central role determining chromatographic retention behavior. Conformational effects choices column temperature type vs hydrophobicity) important tools, which manipulate structure, variant separation. The introduction new matrix supports perfusive nonporous particle) has enhanced speed dramatically (from hours few minutes). To enlarge window increase selectivity, mobile-phase additives detergents, metal ions, urea, guanidine hydrochloride, ethylene glycol, glycerol, organic solvents) particularly widely applied.