Anion exchange chromatography method optimization to mAb protein purification

The effects of resins with different ligands and different pH values on human IgG4 protein were investigated by anion exchange chromatography In the first step, a resin screening study with 5 different positively charged ligands from 4 different brands was applied to the largely purified monoclonal antibodies protein using the Protein A capture step. Following this study, the effect of different pH (6-7-8.1) values on an IgG4 protein with an isoelectric point of 6.9 was investigated on a single resin. During the resin screening process, all protein quality analyzes were examined aiming to reach the resin with the most compatible ligand. According to the studies on the effect of pH values on monoclonal antibody protein, when the pH value was higher than 6.9 different fragments of the protein were removed, and it had a direct effect on the protein charge variant. When the protein pH value was studied equal or below to the isoelectric point value with the anion exchange chromatography flow through method was determined that protein recovery was at the maximum value of %92-%98.