Hydrophobic Interaction Chromatography in Continuous Flow-Through Mode for Product-Related Variant Removal

Product-related impurities are challenging to remove during monoclonal antibody (mAb) purification process due to molecular similarity. Frontal chromatography on hydrophobic interaction resins has demonstrated its capability to effectively remove such impurities. However, process improvements geared towards purity level comes as a trade-off with the yield loss. In this work, we present a hydrophobic interaction chromatography process using multicolumn continuous chromatography (MCC) concept and frontal analysis to remove a high prevalence product related impurity. This design uses a two-column continuous system where the two columns are directly connected during product chase step to capture product wash loss without any in-process adjustment. This polish MCC operation resulted in a 10 % increase in yield while maintaining 99 % purity, despite the presence of 20 % product-related impurities in the feed material. One challenge associated with polish MCC design is that the accumulation of the impurities renders a non-steady state recycling. To surmount this issue and ensure a robust process, a mechanistic model was developed and validated to predict multicomponent breakthrough. This model was capable to predict multiple cycle behavior and accounts for increased impurity concentration. Assisted by the model, the optimized operation parameters and conditions can be determined to account for variation in product load quality. The simulated results demonstrate an effective doubling of productivity compared to conventional batch chromatography.