This paper investigates the influence of buffers and additives on the retention time of proteins in ion exchange chromatography (IEX). IEX is a widely used technique for the separation of proteins based on their surface charges, which influence pH. Buffers play a key role in maintaining the final pH conditions and ensuring the reproducibility of protein separation. The choice of buffers and their ionic strength must be matched to the stability and activity of the proteins. Additives, e.g. in the organic solvent, influence the interaction of the proteins with the stationary phase and have specific and non-specific effects on the retention times. Research shows how important the optimisation of these parameters is for efficient protein purification.
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