UV absorbance measurements play an important role in bioprocess development. Yield and purity are often evaluated in terms of peak percentages in analytical SEC or ion-exchange chromatography. Also, industrial chromatography steps are usually controlled based on UV data with pooling decisions according to absorbance thresholds. Model-based process development would make elaborate screening experiments redundant, once the model has been calibrated to the specific process step. So far, absorbance measurements could not be used directly for modeling chromatography steps as the commonly applied models rely on mass or molar concentration. This study presents mechanistic modeling of an industrially relevant chromatography setting without any knowledge of the feed composition. The model equations were rewritten to employ boundary conditions in UV absorbance units, the absorption coefficients were shifted into the isotherm, and standard parameter estimation procedures could be applied. An anion-exchange chromatography case study of a target protein expressed in Escherichia coli and 11 lumped impurity peaks demonstrated practical applicability. The target protein concentration in the feed material was estimated from chromatograms. Using this method, initially unknown feed concentrations can be determined a posteriori for ion-exchange and multimodal chromatography from single-component absorbance curves.
UV absorption-based inverse modeling of protein chromatography
Tobias Hahn, Pascal Baumann, Thiemo Huuk, Vincent Heuveline, Jürgen Hubbuch
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Engineering in Life Sciences