Spiral column configuration for protein separation by high-speed countercurrent chromatography.
Retention of the stationary phase of aqueous-aqueous polymer phase systems is improved by a spiral column configuration which utilizes the radially acting centrifugal force along the spiral pitch to retain the heavier phase in the outer portion and the lighter phase in the inner portion of the spiral channel. For the separation of proteins which has low mass transfer rates, the system needs further modification of the separation channel to interrupt the laminar flow and enhance mixing of the two phases. Two spiral column assemblies were developed, one using a disk with spiral grooves and the other, the spiral tube support which accommodates the multiple spiral layers made from a single piece of fluorinated plastic tubing. In the spiral disk assembly, the best protein separation is achieved by the mixer-settler system which vigorously mixes two phases by vibrating glass beads placed in every other section of barricaded spiral channel, while in the spiral tube assembly the partition efficiency of proteins is enhanced by compressing the tubing to interrupt the laminar flow of the mobile phase. In both systems protein samples were well resolved by choosing the suitable elution modes.
Ito
Yoichiro
Y
Bioseparation Technology Laboratory, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bldg. 10, Room 8N230, 10 Center Drive, Bethesda, MD 20892-1762, USA.
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