Establishment of a CFD-based kL a model in microtiter plates to support CHO cell culture scale-up during clone selection.

Microtiter plates are a common tool for clone selection in biopharmaceutical development. A way of visualizing and evaluating these systems and key processes parameters is the application of Computational Fluid Dynamics (CFD). CFD is a powerful tool for the modelling of hydrodynamics and mass transfer parameters. In this work, CFD was used to determine the specific surface area, the volumetric power input and the oxygen mass transfer coefficient kL a for two different microtiter plates with different scales (100 µL - 5 mL). For this purpose, a new method of predicting the kL a is presented and calibrated with literature data. Scaling effects in shaken microtiter plates are evaluated by comparing two culture volume scales under various operating conditions. To test validity of these models, three different Boehringer Ingelheim Pharma proprietary CHO production cell lines with different growth characteristics were cultivated using the respective microtiter plates under different conditions until limitations in growth and viability were observable. The cell culture data then was compared to different parameters obtained by CFD. The calculated kL a values match the cell culture performance in the 96-deepwell by predicting lowered oxygen transfer with increasing culture volume and decreasing orbital velocity. The same cells behave differently in the 6-deepwell scale. Here, the overall larger shear stress might cause physical stress for the cells. The kL a model predicts overall higher shear rates for this system, supporting the experimental findings. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018.

Prediction and Reduction of the Aggregation of Monoclonal Antibodies.

Protein aggregation remains a major area of focus in the production of monoclonal antibodies. Improving the intrinsic properties of antibodies can improve manufacturability, attrition rates, safety, formulation, titers, immunogenicity, and solubility. Here, we explore the potential of predicting and reducing the aggregation propensity of monoclonal antibodies, based on the identification of aggregation-prone regions and their contribution to the thermodynamic stability of the protein. Although aggregation-prone regions are thought to occur in the antigen binding region to drive hydrophobic binding with antigen, we were able to rationally design variants that display a marked decrease in aggregation propensity while retaining antigen binding through the introduction of artificial aggregation gatekeeper residues. The reduction in aggregation propensity was accompanied by an increase in expression titer, showing that reducing protein aggregation is beneficial throughout the development process. The data presented show that this approach can significantly reduce liabilities in novel therapeutic antibodies and proteins, leading to a more efficient path to clinical studies.