Isolation of Monoclonal Antibody Charge Variants by Displacement Chromatography

Abstract

This unit discusses the important parameters in designing and optimizing a separation of monoclonal antibody (mAb) charge variants from process streams by ion-exchange displacement chromatography, including sample preparation and selection of matrix, column, and appropriate buffer. A protocol is provided for determination of optimal column binding and displacement conditions, including cleaning and regeneration of the displacement columns.

Highly linear pH gradients for analyzing monoclonal antibody charge heterogeneity in the alkaline range: Validation of the method parameters

“…Our next approach, displacement chromatography [17], did not yield a separation of charge variants, under any conditions. This is in stark contrast to the results that Zhang et al [18] have presented previously.…”

“…The buffers and conditions used for displacement chromatography can be found in Table 2. The displacement experiments were performed according to Zhang et al [18] and McAtee and Hornbuckle [17].…”

“…Expell SP1 was chosen as the displacer, as it has proven to be useful in IgG separations [17,18,27]. No separation of charge variants was detectable after analysis with IEF.…”

“…This method has been used successfully for decades [16] and requires only a simple setup. The next approach was displacement CEX [17], which has been shown to be a method capable of high resolution separations of mAb charge variants [18]. With the advent of high performance displacers, this approach was very promising.…”

Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry

“…This is one important argument for using DE for the separation of proteoforms. Thus, it is not surprising, that DE has been applied to the separation of proteoforms of therapeutic proteins successfully [46,[80][81][82][83][84].…”