In this study, the hydrophobicities of newer generation Capto HIC resins were compared to Sepharose High Performance and Sepharose Fast Flow HIC resins. A monoclonal antibody (mAb) and its corresponding antigen-binding fragment (Fab) were loaded on each resin and eluted in a linear salt gradient. The relative hydrophobicity of each resin was determined from the elution volume.
The study generated a complete hydrophobicity map for our hydrophobic interaction chromatography (HIC) resins. Results showed that relative hydrophobicity of a resin is dependent upon sample composition, emphasizing the importance of testing resin hydrophobicity for each biomolecule.
An application example is also presented. Aggregate removal results are compared using Capto Butyl ImpRes and Butyl Sepharose High Performance resins. Capto Butyl ImpRes achieved a similar decrease in aggregate content with the potential to use higher bed heights and increased flow rates.