Design of Experiments–Driven Purification of Anti-Vascular Endothelial Growth Factor Monoclonal Antibodies

Objective: This study demonstrated the successful application of multimodal chromatography (MMC) as an alternative to protein A chromatography for the purification of mAbs with anti-vascular endothelial growth factor properties.Materials and Methods: PROchievA and mAbSelect PrismA resins were used for protein A chromatographic purification, while Capto MMC and Nuvia cPrime resins were used for multimodal chromatographic purification. Each purification study was conducted as the initial capture step for the mAb produced in the Chinese hamster ovary cell culture. The experimental designs for the purification steps were developed using Modde 13 software, and the resulting data were analysed with the assistance of the same software.Results: Purification steps using protein A resins resulted in yields of approximately 87.2%–93%, whereas yields obtained with multimodal resins were approximately 57.8%–65.2%. The multimodal purification step enabled the production of a monoclonal antibody (mAb) with a higher purity compared to the protein A purification step. It also showed superior performance compared to protein A resins in the removal of acidic and basic variants, achieving approximately 10% greater reduction of these variants in a single step and enabling the production of a purer mAb product.Conclusion: This study highlighted the potential of MMC to enhance the purity of mAbs with greater efficiency in removing charge variants. The purified mAbs were confirmed based on their mass values using the sodium dodecyl sulphate–polyacrylamide gel electrophoresis.