Isoelectric chromatofocusing (ICF) has been reported to be capable of separating protein charge variants through the generation of an elution pH gradient on ion exchange matrices. Due to Donnan equilibria, ion-exchange and ion-displacement effects, the elution pH of a protein during ICF is often different from its isoelectric point (pI) determined with electrokinetic methods such as isoelectric capillary electrophoresis (iCE). Through the modulation of ionic strength and pH during ICF, protein elution pH and separation resolution can be adjusted and optimized. This technique was applied to design ICF separations for monoclonal antibodies such that the chromatographic elution profiles are analogous to iCE electropherograms. Using ICF at preparative scales, specific charge variants of interest can be purified and concentrated to allow for further analysis.<br/>

Isoelectric chromatofocusing as an analogous technique to isoelectric capillary electrophoresis for preparation separation of monoclonal antibodies

253rd National Meeting (2017)

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