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 in finding and fine tuning selectivity. High throughput process development in plates is a valuable tool for rapid evaluation of operating conditions for a HIC step.This work will present a high throughput screening study for the development of a capture step for recombinant Green Fluorescent Protein (rGFP) expressed in E. coli. Parallel screening of HIC media and conditions in 96-well plates facilitated the selection of the most promising HIC media and also show how salt type and salt concentration conditions could be tailored to successfully capture rGFP.  Results showed that both binding and elution conditions could be fine-tuned to obtain high protein purity. This approach allows selection of HIC media and elution profiles to be predicted in a fast and effective way, saving both time and sample compared with traditional column screening.

High-throughput screening of HIC media in PreDictor plates for capturing recombinant green fluorescent protein from E. coli

243rd National Meeting (2012)

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