Abstract
Abstract This work addresses the feasibility of scaling-up phenylboronic acid (PBA) chromatography for the purification of human mAbs directly from Chinese Hamster Ovary cell culture supernatants. Column performance optimization regarding superficial velocity and feed volume was performed with the best results being achieved with superficial velocities of 5.1, 15.3 and 25 cm/min in the adsorption, wash and elution steps, respectively. The column volume was scale-up 100 times from 0.4 cm 3 (laboratory scale) to 4, 16 and 40 cm 3 (preparative scale). This 100-fold scale-up was successfully achieved with a recovery yield of 97.7% and a protein purity of 82.6%. Following purification, purified IgG fractions were characterized in terms of isoelectric point, size, CHO proteins and genomic DNA content. Overall results suggest that using ProSep®-PB for mAbs purification is simple, reproducible, robust and scalable without compromising the target molecule integrity and purity.
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