Hydrophobic Interaction chromatography

Hydrophobic interaction chromatography (HIC) is a versatile method for the purification and separation of biomolecules based on differences in their surface hydrophobicity. Proteins and peptides usually sequence hydrophobic amino acids in domains away from the surface of the molecule. However, many biomolecules considered hydrophilic have sufficient hydrophobic groups exposed to allow interaction with hydrophobic ligands attached to the chromatographic matrix. Compared to reversed phase chromatography, the density of the ligand on the matrix is much lower. This feature promotes the high selectivity of HIC, while allowing mild elution conditions to help preserve biological activity.

Hydrophobic interaction between a biomolecule and the matrix is enhanced by high ionic strength buffers. This makes HIC an ideal “next-step” for the purification of materials that have been precipitated with ammonium sulfate or eluted in high salt concentrations during ion exchange chromatography.

Hydrophobic Interaction Products

Selection guide Hydrophobic Interaction pdf

SOURCE™ - Prepacked RESOURCE™ and PE columns and Lab packs

SOURCE chromatography media are based on a hydrophilic matrix made from monodispersed, rigid, polystyrene/dininylbenzene and substituted with different hydrophobic ligands. The 15 µm particle allows fast high resolution purification and is ideal for final polishing in a purification process. SOURCE 15ETH, 15ISO and 15PHE are available in labpacks as well as prepacked in RESOURCE columns. RESOURCE HIC test kit is used to rapidly sceen for the most suitable medium for a specific application. SOURCE 15PHE is also available in a prepacked Tricorn column designed for high resolution at lab scale and for optimization studies when scaling up.

Product and Order information

Sepharose™ High Performance - Prepacked HiLoad™ and HiTrap™ columns

Phenyl Sepharose High Performance (HP) and Butyl Sepharose HP are 34 µm media suitable for intermediate or polishing steps that require high resolution. Both chromatography media are available in lab packs as well as prepacked in convenient HiTrap columns. When increase capacity is required, use prepacked HiLoad columns.

Product and Order information

Sepharose™ Fast Flow - Prepacked HiTrap™, HiScreen™, HiPrep™ columns and Lab packs

Sepharose Fast Flow is based on a matrix of particles with a mean size of 90 µm. The media is well suited for capture or intermediate purification and in large-scale purifications. Hydrophobic ligands coupled to Sepharose Fast Flow are, phenyl, bultyl, butyl-s and octyl. Phenyl Sepharose 6 Fast Flow is available with high or low levels of phenyl substitution differing in selectivity, efficiency and binding capacity. All chromatography media are available in lab-packs. The different media are also available in HiTrap columns as well as in the HiTrap HIC selection kit, to rapidly screen for the most suitable medium for a specific application. When increase capacity is required, use prepacked HiPrep columns. HiScreen HIC media columns are designed for process development work.

Hydrophobic Interaction chromatography

Handbooks

Hydrophobic Interaction and Reversed Phase Chromatography pdf

Protein Purification Handbook pdf