Hydrophobic Interaction Chromatography (HIC)

Hydrophobic interaction chromatography is a useful tool for the purification of proteins, providing a powerful complementary separation step to ion-exchange and gel filtration chromatography. It has also shown much promise as a simultaneous protein purification and refolding step. As hydrophobic interactions are strongest at high ionic strengths, this approach is often used fairly early in downstream bioprocessing, directly after precipitation steps19 when salt levels are high, or in combination with gel filtration or ion-exchange chromatography. Although hydrophobic interaction chromatography is an adsorptive process, the interactions are generally weaker than in affinity and ion-exchange chromatography, thus resulting in minimal structural damage and loss of activity to the target protein. Hydrophobic interaction chromatography is also very useful in the removal of DNA impurities and for purifying DNA-based products.