Progress towards continuous aqueous two-phase extraction via TAPPIR
September 17-21, 2017
At ICB II, we presented Aqueous Two-Phase Extraction (ATPE) as a non-chromatographic alternative for protein purification. We had developed an aqueous two-phase system with inexpensive and biocompatible PEG 1500 or 4000 and ammonium citrate. We purified several enzymes, more specifically a series of dehydrogenases , to near homogeneity after forward extraction into a PEG-heavy top phase at pH > 9 and back extraction into a bottom phase at pH 4-6; in selected cases, we were able to obtain pure protein in the bottom phase without forward extraction into the top phase. Scale-up of the PEG 1500/4000-ammonium citrate to 5-10 L scale still often gave phase separation times of less than five minutes. However, ATPE technology is characterized by complex phase separation and very limited number of separation stages not offering enough separation efficiency. Furthermore, conventional ATPE does not lend itself to continuous operation.
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Andreas S. Bommarius, Bettina R. Bommarius, Juliane Merz, Carolin Stark, and Gerhard Schembecker, "Progress towards continuous aqueous two-phase extraction via TAPPIR" in "Integrated Continuous Biomanufacturing III", Suzanne Farid, University College London, United Kingdom Chetan Goudar, Amgen, USA Paula Alves, IBET, Portugal Veena Warikoo, Axcella Health, Inc., USA Eds, ECI Symposium Series, (2017). https://dc.engconfintl.org/biomanufact_iii/30