Assessing the Degradation of Biopharmaceutical Compounds by Hydrophobic Interaction Chromatography with UV Detection (HIC-UV) (CAT#: STEM-B-0403-CJ)

The principle for protein adsorption to HIC media is complementary to ion exchange and size exclusion chromatography. Sample molecules containing hydrophobic and hydrophilic regions are applied to an HIC column in a high-salt buffer. The salt in the buffer reduces the solvation of sample solutes. As solvation decreases, hydrophobic regions that become exposed are adsorbed by the media. The more hydrophobic the molecule, the less salt is needed to promote binding. Usually a decreasing salt gradient is used to elute samples from the column in order of increasing hydrophobicity. Sample elution may also be assisted by the addition of mild organic modifiers or detergents to the elution buffer.

Biopharmaceutica

1. The protein binds to hydrophobic ligands on the HIC resin in a binding buffer with a high salt concentration. The interaction is reversed when the ionic strength of the buffer is reduced.
2. The deamidation products and succinimides are well separated and quantified.
3. Acquisition of data, peptide map analysis.

• Sample: Peptides, Proteins, Vaccines, Virus-like particles
• Equipment: Hydrophobic Interaction Chromatography (HIC).