As the monoclonal antibody (mAb) expressing mammalian cell cultures have intensified in the last 10 years, new high performance clarification technologies have been developed to address rising cell densities and increasing levels of soluble and of insoluble contaminants. One of such approach that has been adopted by the industry is HCCF fluid harvesting using chromatographic clarification. Such systems utilize AEX fibrous functional media to effectively capture cells, cell debris, DNA, and HCP that are negatively charged at the harvest cell culture pH. The product, typically a high isoelectric point (pI) IgG, flows through such system because it is either neutral or positively charged. We have previously reported on development and applications of such technologies, their performance, scalability, and platformability with respect to mAb manufacturing processes.<br/><br/>As the candidate pipelines expand into non-classical mAbs and non-mAb modalities, one can no longer assume that all products of interest will have high isoelectric points. If one is to continue to utilize chromatographic clarification based on AEX principle, process strategies need to be developed to enable molecules that have middle and low isoelectric points to pass through the AEX bed without causing other deleterious effects. Here we describe such a strategy that enables products within a wide range of pIs to be effectively harvested from CHO cell cultures with high recoveries. We demonstrate that all of the advantages of the chromatographic clarification can be applied to low isoelectric point modalities with minimal modification to the actual clarification process and to the HCCF. Such strategy can be easily deployed at any process scale from discovery to commercial manufacturing and enables platformability of AEX clarification approach beyond classical high isoelectric point mAbs.

Strategies to increase recovery of low isoelectric products during chromatographic clarification of mAb CHO cell cultures

ACS Spring 2022

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