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978-3-8439-2179-4, Reihe Technische Chemie
Fatma Alexia van Winssen
Tunable Aqueous Polymer-Phase Impregnated Resins for Downstream Processing
147 Seiten, Dissertation Technische Universität Dortmund (2014), Softcover, A5
Aqueous Two-Phase Extraction (ATPE) represents a promising unit operation for downstream pro-cessing of biochemical products. The technique provides several advantages such as a biocompatible environment for the extraction of sensitive and biologically active compounds. However, the tendency of some aqueous two-phase systems to form stable emulsions might lead to long phase settling times causing an increased footprint for the required mixer-settler devices or the need for additional equipment such as centrifuges. In this work, a novel approach to improve ATPE for downstream processing applications called ‘Tunable Aqueous Polymer-Phase Impregnated Resins’ (TAPPIR®)-Technology is presented. The technology is based on the immobilization of one aqueous phase inside the pores of an open porous solid support. The second aqueous phase forms the bulk liquid around the impregnated solids. Due to the immobilization of one phase, phase emulsification and phase settling of ATPE are realized in a single step.
In the present work, the TAPPIR®-Technology is evaluated in view of its application in downstream processing. The impregnation of different macroporous glass and ceramic solids was investigated and could be proven to be stable. Within a proof of principle study, the separation of the dye Patent blue V was successfully performed with the TAPPIR®-Technology. In a next step, the application of the TAPPIR®-Technology for the separation of an artificial protein mixture was shown. The same partitioning levels could be reached for the TAPPIR®-Technology as for the classical ATPE setup. Furthermore, the TAPPIR®-Technology was applied for the extraction of laccase from crude Pleurotus sapidus cell culture supernatant. The comparison to the classical ATPE setup showed that the laccase activity partition coefficient and active laccase enrichment as well as the overall activity yield was the same for both, TAPPIR® and ATPE. These results prove the TAPPIR®-Technology to maintain the initially mentioned major advantages provided by ATPE. At the same time, phase immobilization in TAPPIR® eliminates the phase settling step representing a potential limitation of ATPE. Hence, the TAPPIR®-Technology has been proven to be an alternative unit operation for downstream processing of various compounds.