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Abstract Aqueous two phase system(ATPS) provides a gentle, non-denaturing separation environment for proteins, enzymes. While high-speed countercurrent chromatography (HSCCC) is a liquid-liquid partition chromatography which uses centrifugal force to hold the stationary phase and facilities the mobile phase partitioning through the stationary phase, it can produce high separation efficiency with large sample loading capacity. However, the ordinary HSCCC apparatus (Type J ) fails to retention a satisfactory stationary phase of ATPS because of its high viscosity and low interfacial tension. Nevertheless, the later designed cross-axis planetary centrifuge system(X-CPC) can produce a greater lateral force field and enhances significantly the retention of the ATPS stationary phase. This paper gives a review of the application of these CCC techniques with ATPS in the separation of proteins. Meanwhile, new techniques such as pH－peak focusing CCC and dye-ligand affinity CCC and some new CCC column design for improvement of separation efficiency and retention of ATPS stationary phase are introduced in this paper.
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Received: 15 September 2005
Published: 25 February 2006
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