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dc.contributor.authorGuan, YH-
dc.contributor.authorHewitson, P-
dc.contributor.authorvan den Heuvel, RNAM-
dc.contributor.authorZhao, Y-
dc.contributor.authorSiebers, RPG-
dc.contributor.authorZhuang, Y-P-
dc.contributor.authorSutherland, I-
dc.identifier.citationJournal of Chromatography A, ARTN1424, pp. 102 - 110, (2015)en_US
dc.description.abstractManufacturing 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 20.2 m 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 20 ml/min. We then satisfactorily demonstrated the potential for a weekly protein separation and preparation throughput of ca. 11 g 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 1.6 mm 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.en_US
dc.description.sponsorshipThe BBSRC Grant number BB/FOF/206. YHG and YZ also acknowledge financial support by National Basic Research Program (973 Program 2013CB733600).Ms Yan Zhao (School of Mechanical and Power Engineering) has been grateful to a Training Program of Innovation and Entrepreneurship for Undergraduates Grant number X13219 by East China University of Science and Technology. Contribution and/or encouragement to this work from D. Fisher, T. Bunce, Ying-Dong Liu, Zheng-Tao Zhang, Li-Li Baien_US
dc.format.extent102 - 110 (9)-
dc.subjectCounter-current chromatographyen_US
dc.subjectAqueous two-phase systemsen_US
dc.subjectProtein separation and purificationen_US
dc.subjectToroidal columnen_US
dc.subjectScale up chromatography columnen_US
dc.subjectLiquid-liquid chromatographyen_US
dc.titleScale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatographyen_US
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