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Mixed electrolytes in hydrophobic interaction chromatography †
Author(s) -
Müller Egbert,
Vajda Judith,
Josic Djuro,
Schröder Tim,
Dabre Romain,
Frey Tim
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201200704
Subject(s) - chemistry , chaotropic agent , hydrophilic interaction chromatography , salt (chemistry) , hydrophobic effect , isoelectric point , chromatography , salting out , hofmeister series , ammonium sulfate , electrolyte , sodium , solubility , aqueous solution , organic chemistry , high performance liquid chromatography , electrode , enzyme
An essential part of the modulation of protein‐binding capacity in hydrophobic interaction chromatography is the buffer‐salt system. Besides using “single” electrolytes, multicomponent electrolyte mixtures may be used as an additional tool. Both the protein solubility and the binding capacity depend on the position of a salt in the so‐called H ofmeister series. Specific interactions are observed for an individual protein‐salt combination. For salt mixtures, selectivity, recovery, and binding capacity do not behave like for the single salts that are positioned in between the two mixed components in the H ofmeister series, as the continuous correlation would suggest. Thus, finding strategies for mixed salts could potentially lead to improved capacities in hydrophobic interaction chromatography. Mixtures of ammonium sulfate, sodium citrate, sodium sulfate, sodium chloride, sodium acetate, and glycine were used to investigate the binding capacities for lysozyme and a monoclonal antibody on various hydrophobic resins. Resin capacity for two investigated proteins increases when mixtures consisting of a chaotropic and a kosmotropic salt are applied. It seems to be related to the rather basic isoelectric points of the proteins.