Comparison of various columns used in anion exchange chromatography method for mAb purification

Abstract

The impact of resins with varying ligands and pH levels on human IgG4 protein was analysed using anion exchange chromatography. Initially, a resin screening study involving five different positively charged ligands from four different brands was conducted on largely purified monoclonal antibodies, following Protein A capture. Subsequently, the influence of pH levels (6, 7, and 8.1) on an IgG4 protein with an isoelectric point of 6.9 was assessed using a single resin. Throughout the resin screening, all protein quality analyses were performed to identify the resin with the most compatible ligand. The study on pH effects revealed that when the pH exceeded 6.9, various protein fragments were removed, directly affecting the protein charge variant. When the protein pH was at or below the isoelectric point, the anion exchange chromatography flow-through method achieved a maximum protein recovery of 92-98%.

Keywords

• Anion exchange chromatography
• Monoclonal antibody
• Igg
• Resin screening
• Downstream

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