Hydrophobic interaction chromatography (HIC) is a powerful purification technique where the type and density of the ligand, pH and salt of binding conditions, temperature and the nature of the target protein are highly significant parameters when determining selectivity. This study demonstrates a rapid, high-throughput process development workflow using parallel screening in 96-well plates to determine the most suitable HIC media and conditions for capturing recombinant Green Fluorescent Protein expressed in E. coli. Chromatography media were ranked and their elution profiles predicted in a fast and effective way, saving both time and sample compared with traditional column screening. Results showed that binding and elution conditions could be fine-tuned to obtain high protein purity.
September 5, 2011