Ion exchange (IEC) and multi-modal chromatography (MMC) are widely used methods for the removal of contaminants like host cell proteins, aggregates or charged variants from therapeutic proteins.Classical elution modes in ion exchange chromatography include linear salt gradients or step elutions at a fixed pH value. Use of pH as a mobile phase modifier (pH gradients) in IEC can be an effective alternative to using salt gradients. Advantages of pH gradients include the possibility of greater resolving power and elution at low ionic strengths.In multi-modal chromatography often a combination of pH and salt gradients or steps for elution is necessary for high resolution and recovery rates.In this work we present data for the separation of monoclonal antibodies, rec. proteins and peptides by externally generated dual linear salt and pH gradients, which is used for modeling and prediction of the elution behavior.The analysis is based on the linear gradient elution model of Yamamoto and allows the calculation and prediction of the elution behavior of proteins for dual linear salt and pH gradient in IEC and MMC.

Modeling of dual gradient chromatofocusing in ion exchange and multi-modal chromatography

247th National Meeting (2014)

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