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. TSKgel BioAssist Phenyl columns contain the same particles as Phenyl-5PW columns, but are packed in PEEK column hardware rather than stainless steel. BioAsist Phenyl (Phenyl-5PW), TSKgel Ether-5PW and Butyl-NPR resin-based columns provide a range of hydrophobicities for chromatographic optimization. Ammonium sulfate concentration can be minimized by using a more hydrophobic packing, such as BioAssist Phenyl. We recommend an Ether-5PW column for purifying very hydrophobic proteins. TSKgel BioAssist Phenyl and TSKgel Ether-5PW packings are based on TSKgel G5000PW resin - 10 µm particles with 1000 Ã pores. TSKgel Butyl-NPR columns, which are less hydrophobic than Ether-5PW, is prepared from 2.5 µm 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.

The most hydrophobic among the three TSKgel HIC columns, TSKgel Phenyl-5PW is an excellent choice to screen for the selectivity, retention and recovery of most biomolecules. Featuring a 1000Å pore size in a polymethacrylate base material, the Phenyl-5PW column is stable in either acid or caustic cleaning regimens and provides excellent access to larger molecules with low diffusion coefficients. Stainless steel is the standard column format for TSKgel Phenyl-5PW. Glass and PEEK column housings are available for samples that may adsorb to stainless steel. The alternative column formats are also suitable for general applications.

Legal Information

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