Development of an intermediate chromatography step in an insulin purification process. The use of a High Throughput Process Development approach based on selectivity parameters.

Recent innovations in designing purification processes for biopharmaceutical production have enabled initial screening (optimization) of chromatographic conditions for binding to be performed in miniaturized batch format. The present report demonstrates the possibility of using this format to screen for selectivity and illustrates the need for careful adjustment of protocols when highly abundant, tightly-binding impurities are present in the sample. This batch format approach was used to choose a chromatography medium (resin) from a selection of available resins for the purification of recombinant insulin expressed in E.

Characterization of multimodal hydrophobic interaction chromatography media useful for isolation of green fluorescent proteins with small structural differences.

Hydrophobic interaction chromatography (HIC) has been developed as a powerful technique for separating and purifying proteins. In this study, we have characterized the ability of new multimodal pH-HIC media to resolve proteins with only small differences in their primary structures. This was done by determining the retention times of different green fluorescent protein (GFP) mutants prepared from Escherichia coli extracts.