Selection and Use of Poly Ethylene Glycol and Phosphate Based Aqueous Two-Phase Systems for the Separation of Proteins by Centrifugal Partition Chromatography

Separation of active laccases from Pleurotus sapidus culture supernatant using aqueous two-phase systems in centrifugal partition chromatography

“…It has been shown in earlier studies [24] that the retention of stationary phase for ATPS composed of PEG and phosphate salts was better when using the PEG rich upper phase as stationary phase. Thus, the descending mode of operation was chosen where the lower salt rich phase of the ATPS is the mobile phase.…”

“…Hydrodynamic measurements in the CPC chamber were performed to determine the volume flow of mobile phase for the separation experiments. Detailed information about hydrodynamic measurement and the experimental setup of a single disc CPC is given in a former publication [24].…”

“…Six-port valves (Knauer, Berlin, Germany; type: Smartline S6, 3-channel) were used to select the phase to be pumped to the CPC and for sample injection. The flow out of the CPC was cooled down by a steel tube in iced water before UV-detection to decrease the noise in the detector baseline as described in a former study [24]. For UVdetection a ProStar 340 detector (Agilent, Santa Clara (CA), USA) at 210 nm and a fraction collector L-7655 (Merck-Hitachi, Darmstadt, Germany) was used.…”

“…The rotor is built up of 20 discs, containing 1320 chambers and 200 mL internal volume in total. The peripheral setup is as described in further studies [24,25]: solvent supply is done by two SD-1 pumps (Agilent, Santa Clara (CA), USA), one for each phase. Six-port valves (Knauer, Berlin, Germany; type: Smartline S6, 3-channel) were used to select the phase to be pumped to the CPC and for sample injection.…”

Developing the biofacility of the future based on continuous processing and single-use technology

“…Consequently, alternative UO for the capture step or intermediate purification/polishing like aqueous two phase extraction (Muendges et al, 2015;Rosa et al, 2011;Schwienheer et al, 2015), precipitation (Sim et al, 2012;Hammerschmidt et al, 2014) or crystallization (Schmidt et al, 2005) have not been taken into account. Although, these methods might have advantages regarding continuous operation mode and process economics, they do not fulfill the requirement of being a platform technology in antibody production.…”