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978-3-8439-2333-0, Reihe Verfahrenstechnik

Jan Mündges
Investigation and Intensification of Monoclonal Antibody Purification by Aqueous Two-Phase Extraction

197 Seiten, Dissertation Technische Universität Dortmund (2015), Softcover, A5

Zusammenfassung / Abstract

The application of monoclonal antibodies (mAbs) for therapeutic treatment of several diseases like cancer, asthma or autoimmune diseases causes a high demand for mAbs. Continuous manufacturing of mAbs offers the opportunity to increase productivity and reduce apparatus dimensions. Aqueous two-phase extraction (ATPE) is a promising technology to replace or support chromatographic process steps, especially Protein A chromatography, and lead to a continuous downstream process.

Within this work a continuous, multistage ATPE process for the purification of an Immunoglobulin G1 (IgG1) from a Chinese hamster ovary cell supernatant is developed. For process development a combination of experimental investigation and process modeling is applied. The result is a continuous, multistage ATPE process which combines a salt-rich extraction step, followed by a salt-poor back extraction and a salt-poor washing step. IgG1 yields and purities achieved are in the range of Protein A chromatography performance and underline the potential of the ATPE process. Nonetheless, precipitation of proteins is observed to be a significant drawback. Parallel, a process model is developed to predict the multistage ATPE on the basis of single stage equilibrium data is successfully validated by the experimental results. The model represents a useful tool for future ATPE process development and significantly decreases experimental efforts.

ATPE was further successfully applied as a pre-concentration step to Protein A chromatography. In a single stage batch extraction IgG1 is significantly concentrated directly from cell culture. Because of the limited loading capacity of aqueous two-phase systems a significant amount of IgG1 is lost due to occurring precipitation. By modifying the aqueous two-phase system with hyperbranched polymers the loading capacity was increased 30 fold. This work therefore significantly contributes to the future application potential of ATPE.