Protein separation using toroidal columns by type-J synchronous counter-current chromatography towards preparative separation文档.pdfVIP

Journal of Chromatography A, 1217 (2010) 3525–3530 Contents lists available at ScienceDirect Journal of Chromatography A j o u rn al h omepage: w w w. elsev /locate/chro ma Protein separation using toroidal columns by type-J synchronous counter-current chromatography towards preparative separation Y.H. Guan ∗ , D. Fisher, I.A. Sutherland Brunel Institute for Bioengineering, Brunel University West London, Uxbridge, Middlesex UB8 3PH, UK a r t i c l e i n f o a b s t r a c t Article history: Separation of large bioactive molecules such as proteins, DNAs and RNAs using aqueous two-phase sys- Received 2 October 2008 tems (ATPSs) and liquid–liquid partition-based counter-current chromatography (CCC) can avoid risks of Received in revised form 13 March 2010 sample loss and denaturation, and greatly reduce processing time. We have constructed toroidal columns Accepted 22 March 2010 (length 26–140 m, column volume 51–280 ml, bore size 1.6 mm) suitable for mounting onto a commer- Available online 27 March 2010 cially available preparative CCC apparatus. With the use of an ATPS containing 12.5% (w/w) PEG1000 and 12.5% (w/w) K2 HPO4 and at a rotational speed of 800 rpm for the rotor of the CCC device, the lower phase Keywords: (i.e. the phosphate-enriched phase) has been used as the mobile phase and a pair of proteins, myoglobin Counter-current chromato