Hydrophobic Interaction Chromatography

Hydrophobic Interaction Chromatography

Hydrophobic interaction chromatography (HIC) is driven by non-polar interactions between a non-polar stationary phase and molecular hydrophobicity exposed by a high salt concentration mobile phase. Chaotropic salts in the mobile phase significantly alter hydration patterns around analytes in solution, forcing hydrophobic regions to reorganize and variably interact with a weakly non-polar stationary phase. Analytes generally elute in order of most hydrophilic to most hydrophobic, facilitated by decreasing salt concentration in the mobile phase. Low percentages of organic solvent in the mobile phase can be utilized to facilitate elution for strongly binding species.

Applications

HIC is generally considered a non-denaturing separation technique for biologics analysis, typically preserving tertiary and quaternary-structural interactions within proteins, antibodies, and nucleic acids. It is applied for protein separations, protein purifications, DNA and RNA isoform analyses, viral clearance for AAVs, analysis of monoclonal (mAbs) and bispecific antibodies (bsAbs), and analysis of Antibody-Drug Conjugates (ADCs).

HIC Solutions

TSKgel^® HIC columns contain polymer-based stationary phases linked with a variety of hydrophobic ligands suitable for protein, DNA, and antibody-based applications.

TSKgel^® HIC-ADC columns contain polymer-based stationary phases with novel grafting chemistries targeted specifically for resolving structural features of variably conjugated ADC isoforms.