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How does Hydrophobic Interaction Chromatography (HIC) work?
Posted May 8, 2024

Answer

In HIC, proteins are inserted into a column with a high-salt buffer. The salt reduces solvation around the proteins, exposing their hydrophobic regions and promoting interaction with the medium. The amount of salt required for binding depends on the protein's hydrophobicity, with less hydrophobic proteins requiring more salt. Elution is achieved by decreasing the salt concentration, allowing proteins to be released based on their increasing hydrophobicity. Bound proteins can be washed off using a dilute buffer or water.