<br/>A key aspect of bioprocess chromatography process design is maximizing product quality attributes by optimizing process parameters and performance attributes. Process parameters include but are not limited to bed height, elution pH, and eluate cut points. An under characterized chromatography process parameter for protein A chromatography is process temperature. Here, we present the effects of controlling temperature (10, 20, and 30°C) on the purification of a monoclonal antibody (mAb) from a perfusion bioreactor using MabSelect SuRe (Cytiva, Marlborough, MA, US) chromatography resin. Batch adsorption experiments using 96-well chromatography resin plates incubated at the three different temperatures generated the isotherms, thermodynamic parameters, and dynamic binding capacities (DBCs) of the columns. Numerical and shrinking core mechanistic models fit to the DBC data provided the mass transfer and diffusion coefficients used to predict column behavior when scaling up to 1 mL columns. A self-designed 3D-printed heating jacket controlled the 1 mL chromatography process temperature during the loading, wash, elution, and cleaning in place (CIP) steps. The numerical and mechanistic model predictions were compared with the scale-up results to determine the effectiveness of each model to predict scale-up behavior. Furthermore, the results at 10°C demonstrated a 20% increase in binding capacity and a 35% reduction in mAb loss during the loading and wash phases, respectively. In addition, eluting at 30°C resulted in the formation of a second early eluting peak which contained 60% of the total aggregates and dimers and only 7% of the total mAb bound to the column. Removing this early eluting impurity-rich peak offers the potential to simplify further polishing steps by reducing the impurity content in the pooled product. The addition of temperature control to protein A chromatography purification provides a potentially novel method to enhance yields and purity whilst reducing resin volume required.

Effect of temperature on the operational efficiency of protein A chromatography for the purification of a monoclonal antibody

ACS Spring 2022

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