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J Chromatogr A. 2015 Dec 11;1424:102-10. doi: 10.1016/j.chroma.2015.03.004. Epub 2015 Mar 11.

Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography.

Author information

1
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.
2
Institute of Environment, Health and Societies, Brunel University London, Uxbridge, Middlesex UB8 3PH, UK.
3
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.
4
Department of Applied Physics, Fontys University of Applied Sciences, Rachelsmolen 1, 5600 AH Eindhoven, The Netherlands.

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.

KEYWORDS:

Aqueous two-phase systems; Counter-current chromatography; Liquid–liquid chromatography; Protein separation and purification; Scale up chromatography column; Toroidal column

PMID:
25818556
DOI:
10.1016/j.chroma.2015.03.004
[Indexed for MEDLINE]

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