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

Abstract

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.

Model-Based Design and Process Optimization of Continuous Single Pass Tangential Flow Filtration Focusing on Continuous Bioprocessing

“…For the biotechnological and pharmaceutical industry membrane-based unit operations are key process steps in order to adjust the concentration of the target component and the buffer media composition. This is mainly based on their mild temperature and process conditions and the low operation costs [1][2][3][4][5][6][7][8][9][10][11]. A high concentration of target protein leads to improved operating conditions for the following separation steps, e.g., chromatography, and lowers their operation costs.…”

Evaluation of Continuous Membrane Chromatography Concepts with an Enhanced Process Simulation Approach

“…The static binding capacity was measured with batch adsorption measurements in a way very similar to the one described by Schwellenbach et al [28]. For the simulation of chromatography in bind and elute mode, the isotherms have to be measured with respect to the modifier concentration.…”

“…Besides different unit operations, there is a huge potential for chromatographic separations that are not particle-based, such as membrane- or fiber-based chromatography [28,29,30,31,32,33,34].…”

Streamlined process development procedure incorporating the selection of various stationary phase types established in a mAb aggregate reduction study with different mixed mode ligands

“…The scaffold can be compared to monoliths and membrane material as, for example, polyethylene terephthalate and cellulose. [36][37][38] Membrane adsorber backbone structures with its pore sizes of 2-4 μm are known for their high applicable flow range with relatively low pressure drop. 39 These membrane adsorber are usually made of organic material such as cellulose and its derivatives, polyethersulfone, polypropylene and polyvinylidenfluoride exhibiting a surface are of 0.6-3 m 2 /g.…”