کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
640019 | 1456960 | 2016 | 8 صفحه PDF | دانلود رایگان |
• Direct capture of monoclonal antibodies from a serum-free culture medium by PBA chromatography.
• 100 times scale-up successfully accomplished.
• High clearance of host DNA and proteins.
• Column performance optimization performed regarding superficial velocity and feed volume.
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 cm3 (laboratory scale) to 4, 16 and 40 cm3 (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.
Journal: Separation and Purification Technology - Volume 160, 29 February 2016, Pages 43–50