Purification of Monoclonal Antibodies Using a Fiber Based Cation-Exchange Stationary Phase: Parameter Determination and Modeling

Bioengineering

Steffen Zobel¹  Jochen Strube¹  Louis Villain²  Jan Schwellenbach²  Florian Taft² 

[1] Institute for Separation and Process Technology, Clausthal University of Technology, Clausthal-Zellerfeld 38678, Germany;Sartorius Stedim Biotech GmbH, Göttingen 37079, Germany;

关键词: monoclonal antibodies;    downstream processing;    cation-exchange chromatography;    model;    simulation;    parameter determination;   

DOI  :  10.3390/bioengineering3040024

来源: DOAJ

【 摘 要 】

Monoclonal antibodies (mAb) currently dominate the market for protein therapeutics. Because chromatography unit operations are critical for the purification of therapeutic proteins, the process integration of novel chromatographic stationary phases, driven by the demand for more economic process schemes, is a field of ongoing research. Within this study it was demonstrated that the description and prediction of mAb purification on a novel fiber based cation-exchange stationary phase can be achieved using a physico-chemical model. All relevant mass-transport phenomena during a bind and elute chromatographic cycle, namely convection, axial dispersion, boundary layer mass-transfer, and the salt dependent binding behavior in the fiber bed were described. This work highlights the combination of model adaption, simulation, and experimental parameter determination through separate measurements, correlations, or geometric considerations, independent from the chromatographic cycle. The salt dependent binding behavior of a purified mAb was determined by the measurement of adsorption isotherms using batch adsorption experiments. Utilizing a combination of size exclusion and protein A chromatography as analytic techniques, this approach can be extended to a cell culture broth, describing the salt dependent binding behavior of multiple components. Model testing and validation was performed with experimental bind and elute cycles using purified mAb as well as a clarified cell culture broth. A comparison between model calculations and experimental data showed a good agreement. The influence of the model parameters is discussed in detail.

【 授权许可】

Unknown