General description

Both hydrophobic interaction chromatography (HIC) and reversed-phase liquid chromatography separate on the basis of protein hydrophobicity and allow selective binding and desorption of proteins. However, HIC operates at significantly lower binding energy and uses aqueous mobile phases. These characteristics provide a gentle technique that is less likely to disturb protein conformation. As a result, HIC generally provides better activity recovery.Three resin-based TSKgel HIC columns provide a range of hydrophobicities for chromatographic optimization. TSKgel Phenyl-5PW columns are most hydrophobic, followed by Ether-5PW, while the surface of Butyl-NPR columns is most polar or least hydrophobic. Ammonium sulfate concentration can be minimized by using a more hydrophobic packing such as Phenyl-5PW. We recommend the more polar Ether-5PW column for purifying more hydrophobic proteins. Ether-5PW and Phenyl-5PW packings are based on TSKgel G5000PW resin - 10 micron particles with 1000 àpores. Butyl-NPR packing is prepared from 2.5 micron nonporous particles, allowing for rapid analyses and semi-micropreparative applications. All three column types can be cleaned with 0.2 M NaOH. Scale-up can be performed by using corresponding Toyopearl bulk resins.

Legal Information

TSKgel is a registered trademark of Tosoh Corporation

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