title: Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography.
abstract: Manufacturing high-value added biotech biopharmaceutical products (e.g. therapeutic proteins) requires quick-to-develop, GMP-compliant, easy-to-scale and cost effective preparatory chromatography technologies. In this work, we describe the construction and testing of a set of 5-mm inner diameter stainless steel toroidal columns for use on commercially available preparatory scale synchronous J-type counter-current chromatography (CCC) machinery. We used a 20.2m long column with an aqueous two-phase system containing 14% (w/w) PEG1000 and 14% (w/w) potassium phosphate at pH 7, and tested a sample loading of 5% column volume and a mobile phase flow rate of 20ml/min. We then satisfactorily demonstrated the potential for a weekly protein separation and preparation throughput of ca. 11g based on a normal weekly routine for separating a pair of model proteins by making five stacked injections on a single portion of stationary phase with no stripping. Compared to our previous 1.6mm bore PTFE toroidal column, the present columns enlarged the nominal column processing throughput by nearly 10. For an ideal model protein injection modality, we observed a scaling up factor of at least 21. The 2 scales of protein separation and purification steps were realized on the same commercial CCC device.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, & The College of Biotechnology, East China University of Science & Technology, Shanghai 200237, China; Shanghai Collaborative Innovation Center for Biomanufacturing Technology, 130 Meilong Road, Shanghai 200237, China. Electronic address: y.h.guan@ecust.edu.cn.
authors.forename: Yue Hugh
authors.initials: YH
authors.lastname: Guan
authors.affiliation: Institute of Environment, Health and Societies, Brunel University London, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Hewitson
authors.affiliation: Institute of Environment, Health and Societies, Brunel University London, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Remco N A M
authors.initials: RN
authors.lastname: van den Heuvel
authors.affiliation: State Key Laboratory of Bioreactor Engineering, & The College of Biotechnology, East China University of Science & Technology, Shanghai 200237, China; Shanghai Collaborative Innovation Center for Biomanufacturing Technology, 130 Meilong Road, Shanghai 200237, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Applied Physics, Fontys University of Applied Sciences, Rachelsmolen 1, 5600 AH Eindhoven, The Netherlands.
authors.forename: Rick P G
authors.initials: RP
authors.lastname: Siebers
authors.affiliation: State Key Laboratory of Bioreactor Engineering, & The College of Biotechnology, East China University of Science & Technology, Shanghai 200237, China; Shanghai Collaborative Innovation Center for Biomanufacturing Technology, 130 Meilong Road, Shanghai 200237, China.
authors.forename: Ying-Ping
authors.initials: YP
authors.lastname: Zhuang
authors.affiliation: Institute of Environment, Health and Societies, Brunel University London, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Sutherland
mesh_terms: D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods; D007202:Indicators and Reagents; D008962:Models, Theoretical; D009113:Muramidase / Q000302:isolation & purification; D009211:Myoglobin / Q000302:isolation & purification; D010710:Phosphates; D011092:Polyethylene Glycols; D017680:Potassium Compounds; D011506:Proteins / Q000302:isolation & purification*; D013193:Stainless Steel
chemical_list: D007202:Indicators and Reagents; D009211:Myoglobin; D010710:Phosphates; D017680:Potassium Compounds; D011506:Proteins; D013193:Stainless Steel; D011092:Polyethylene Glycols; C013216:potassium phosphate; D009113:Muramidase; C000595209:polyethylene glycol 1000
keywords: Aqueous two-phase systems; Counter-current chromatography; Liquid–liquid chromatography; Protein separation and purification; Scale up chromatography column; Toroidal column