This presentation illustrates pro-insulin purification using chromatographic media (resins) coupled with different multimodal cation ligands. To compare the ligands, different binding and elution conditions were tested for each structure.The evaluation was performed using a High Throughput Process Development (HTPD) technology, using media filled 96-well filter plates. The sample used in the study was a recombinant pro-insulin expressed in E.Coli. Binding of pro insulin to the different ligands were tested at different pH and elution was performed at different pH and sodium chloride concentrations. To keep the pro insulin dissolved all experiments were run in 8M Urea. The HTPD results of both binding and elution studies show that the ligand structure will have an important role for the pro insulin purification. The pro insulin binds stronger to some ligands and weaker to others. Also ability to elute varied with different kind of ligands.This presentation shows the HTPD results, how different kinds of ligands and running conditions affect pro insulin purification results.

Evaluation of different cation multimodal ligand structures for capture of pro insulin

245th National Meeting (2013)

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