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ISBN 978-3-8439-2916-5

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978-3-8439-2916-5, Reihe Technische Chemie

Christoph Heinrich Schwienheer
Advances in centrifugal purification techniques for separating (bio-) chemical compounds

234 Seiten, Dissertation Technische Universität Dortmund (2016), Softcover, A5

Zusammenfassung / Abstract

Increasing production of biochemical compounds causes a demand for benign purification techniques. Chromatography is capable of achieving highly purified products, however, biochemical compounds like active enzymes can irreversibly adsorb at the solid stationary phase. Centrifugal partition Chromatography (CPC) is a technique without the use of a solid stationary phase and thus, potentially provides benign conditions. Therefore, one liquid phase is kept stationary in chambers on a rotor with a centrifugal field, while a second immiscible liquid mobile phase is pumped through. Compounds distribute between both phases and elute time-dependent according to their affinity to the liquid stationary phase. CPC is commonly operated with aqueous-organic solvent systems, however, these are mostly not suitable for biochemical compounds, as protein structures might get destroyed. Aqueous two-phase systems (ATPS) provide benign conditions for biochemical compounds. By using ATPS in CPC, a chromatographic separation technique for biochemical compounds arises. However, the use of ATPS in CPC is not well investigated yet and efficiency is low. The scope of this thesis is therefore to enhance the separation efficiency by investigating the hydrodynamic, namely the effects of the two-phase flow in the CPC chambers. By this, and the connection to the mass transfer, parameters influencing the separation efficiency are found. As a result the separation performance was increased by new chamber designs, allowing to be operated at higher rotational speed, resulting in increased interfacial area for mass transfer. With the use of ATPS, which were systematically investigated in the CPC, the knowledge was transferred to the purification of biochemical compounds. Separations experiments were performed with model proteins and enzymes. Activity yields of 100 % demonstrated the benign and activity preserving combination of ATPS in CPC. Finally, a concept for a continuous operating extraction device, based on the CPC technology was phrased and a first prototype built.