Scale up of a chromatographic capture step for a clarified bacterial homogenate – Influence of mass transport limitation and competitive adsorption of impurities

https://doi.org/10.1016/j.chroma.2020.460856Get rights and content
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Highlights

  • Model-based approach for scaling up chromatographic capture step was developed.

  • Model system for the study was fibroblast growth factor protein from an E. coli homogenate.

  • Dynamic model was formulated which was used to scale up the operation.

  • Effects of feed viscosity and competitive adsorption of impurities were accounted for.

Abstract

A model-based approach for scaling up chromatographic capture step was developed. The purification of human basic fibroblast growth factor protein 2 (FGF2) from an E. coli homogenate on a cation exchange resin was selected as a case study. Non-ideal effects accompanying the capture operation were examined, including: reduction in the protein diffusivity in the presence of the homogenate, competitive adsorption between FGF2 and undefined impurities, and flow behavior in external column volumes. The viscosity of the homogenate was measured as a function of dilution degree and shear stress, and its contribution to the diffusivity reduction was quantified. A dynamic model was formulated which accounted for underlying kinetic and thermodynamic dependencies. The model parameters were determined for a lab scale system using a small 2-mL column. The model was successfully used to scale up the capture operation from the lab scale column to a preparative bench scale column of about 1 L volume.

Keywords

Mathematical modelling
Protein capture
Scaling up
Fibroblast growth factor

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