Removing aggregates during the downstream purification of monoclonal antibody (mAb) therapeutics is imperative since these impurities increase the risk of an immunogenic response and can reduce efficacy. Aggregates are a particularly challenging impurity to remove since they have very similar isoelectric points and hydrophobicities to the monomeric protein and are typically present at much higher percentages relative to other impurities such as HCP, DNA, viruses, etc. Aggregate removal is most commonly performed using bind/elute chromatographic processes. Here, we report a unique CEX resin specifically designed for the removal of aggregates in the flow-through frontal chromatography mode of operation, thereby enabling significantly higher loadings (1000 g/L) relative to bind/elute chromatography (30-80 g/L). In this study, we compare two different 3-step processes consisting of capture chromatography with Eshmuno® A media, followed by CEX bind/elute chromatography with Eshmuno® CPX media or CEX flow-through frontal chromatography with a new CEX media, and finally flow-through chromatography with Eshmuno® Q. We demonstrate comparable aggregate and HCP removal with good recoveries for both processes using two different mAbs. However, the process using the CEX flow-through frontal chromatography was operated at greater than 10 times higher loadings and requires no dilution prior to AEX chromatography, providing a purified product at 2.5 times the concentration obtained with a traditional CEX bind/elute chromatography process.

MAb aggregate removal with CEX flow-through frontal chromatography

255th National Meeting (2018)

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