Taper Spiral Column For Protein Separation By Type-J Synchronous Counter-current Chromatography

Countercurrent chromatography(CCC)takes advantage of physical forces,typically the Archimedean screw force and centrifugal force,to retain a liquid stationary phase in a column while undergoing planetary motion,and a mobile immiscible liquid phase flows over the stationary phase to achieve liquid-liquid partition.Samples or crude materials usually are injected into the mobile phase,after the numerous mixing and settling steps a chromatographic separation is obtained based on the different distribution coefficients of the sample components.Thus CCC haves many advantages over other chromatographic methods in that it eliminates complications caused by the use of solid supports,such as irreversible adsorptive sample deactivation and loss,tailing of solute peaks,and contamination.So far,multi-layer helical columns on J-type CCC apparatus has been widely used for the separation of various natural and synthetic products.However,this type of columns on a J-type CCC apparatus did not work as well for protein separation using aqueous two-phase systems(ATPSs).So it limits the CCC's applications in the separation of biologically active macromolecules.This paper,firstly,minutely summarizes principle,apparatus,and methodology of the countercurrent chromatography.Secondly,we elaborate our physical model for "spiral"column on synchronous Type-J CCC planetary motion while briefly stating the conventional physical model for "ring" columns which reveals certain key features of helical and multi-layer helical columns.By comparing the "spiral" column with the "ring" column,we can get that the spiral model is more rationality and more advanced."ring" model was developed for circular columns and in practice this refers to single-layer helical columns and,to a large extent,multi-layer helical ones as well.Our spiral model has been developed for columns with spiral geometry.Thirdly,based on our "spiral" model,we designed a tapered multi-layer spiral columns.In the end,we use the tapered multi-layer spiral columns on synchronous Type-J CCC planetary motion to separate proteins,and it is successful to finish the separation process.No doubt,our research findings is great important to apply such scalable CCC devices to separation and purification of large bioactive molecules.