Control of antibody high and low molecular weight species by depth filtration‐based cell culture harvesting. Issue 10 (29th June 2019)

• Record Type: Journal Article
• Authors: Yu, Deqiang; Mayani, Mukesh; Song, Yuanli; Xing, Zhizhuo; Ghose, Sanchayita; Li, Zheng Jian
• Is Part Of: Biotechnology and bioengineering. Volume 116:Issue 10(2019)
• Page Start: 2610
• Page End: 2620
• Publication Date: 2019-06-29
• Subjects: cell culture harvesting -- depth filtration -- high molecular weight species -- low molecular weight species -- monoclonal antibody -- protein adsorption; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.27081
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Depth filtration‐based harvesting is widely used in mAb manufacturing to remove cell and process‐related impurities. However, it has not been studied on control of product‐related impurities, which are very critical for product quality. In this article, we studied the interactions of depth filter with high and low molecular weight species (HMWs and LMWs) for their direct removal from cell culture. The process parameters (filter, loading, temperature, and flux) were evaluated for adsorption of HMWs and LMWs by depth filters. The adsorption is significantly dependent on filter media and loading capacity and is mainly on the basis of hydrophobic interaction during harvesting. The HMW and LMW species were characterized as HMW1, HMW2, LMW1, and LMW2. The increasing binding from LMW2 to LMW1, HMW1, and HMW2 is correlated with their increasing hydrophobicity score. Adsorption using enriched HMW sample demonstrated similar total protein binding capacity (36–40 g/m 2 ) between depth filters D0HC and X0HC. However, X0HC has stronger HMW binding than D0HC (71% vs 43% of bound protein), indicating more hydrophobic interaction in X0HC. HMW2 DBC on X0HC reached 12 g/m 2, similar to protein binding on hydrophobic interaction membrane adsorbers. Further study showed LMW can induce HMW formation. This study provides a critical understanding of HMW and LMW interaction with depth filters. The strategy of HMW and LMW control by depth filtration‐based harvesting was implemented successfully in mAb manufacturing. Abstract : A novel strategy was developed in this study to directly control antibody high and low molecular weight species (HMW and LMW) during cell culture harvesting by depth filtration. Deqiang and coworkers studied the interaction of HMW and LMW species with depth filters by characterizing protein species and properties. The process parameters and binding capacities were optimized for adsorption of HMW and LMW and this strategy was successfully implemented in mAb manufacturing.