Progress towards continuous aqueous two-phase extraction via TAPPIR

Conference Dates

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 [1], 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.[2] 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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