Optimization Of A Hydrophobic Interaction Chromatography Step For Recombinant Protein Purification

By Eva Heldin, BioProcess R&D Section Leader, Cytiva

Explore a workflow employed for the purification of a recombinant protein expressed in E. coli utilizing hydrophobic interaction chromatography (HIC). This workflow includes the use of high-throughput process development (HTPD) plates to facilitate the choice of resin and initial conditions for column chromatography. The range of buffer conditions utilized in this study was based on the results of an initial solubility study, and all the resins used were Capto™ high productivity resins. Based on the chromatographic results, we found that a wash step before the elution step reduced the amount of impurities. We also performed a design of experiments (DoE) to optimize the wash step and the results were confirmed in a verification run.