title: Spiral column configuration for protein separation by high-speed countercurrent chromatography.
abstract: 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.
authors.affiliation: 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.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Ito
references.citation: J Chromatogr A. 2009 May 8;1216(19):4193-200
references.pmid: 19062024
references.citation: J Chromatogr A. 2003 Oct 31;1017(1-2):71-81
references.pmid: 14584692
references.citation: J Chromatogr A. 2007 Jun 1;1151(1-2):115-20
references.pmid: 17239387
references.citation: J Chromatogr. 1984 Oct 5;301(2):387-403
references.pmid: 6501496
references.citation: J Chromatogr A. 1998 Apr 10;802(2):277-83
references.pmid: 9595695
references.citation: J Chromatogr A. 2005 Feb 18;1065(2):145-68
references.pmid: 15782961
references.citation: J Liq Chromatogr Relat Technol. 2008 Jan 1;31(9):1346-1357
references.pmid: 19343107
references.citation: Science. 1975 Sep 19;189(4207):999-1000
references.pmid: 1220011
references.citation: J Chromatogr. 1984 Oct 5;301(2):405-14
references.pmid: 6501497
references.citation: J Chromatogr A. 2007 Nov 23;1172(2):151-9
references.pmid: 17961580
references.citation: J Chromatogr A. 2007 Jun 1;1151(1-2):108-14
references.pmid: 17316660
references.citation: Science. 1970 Jan 16;167(3916):281-3
references.pmid: 5409709
references.citation: Anal Chem. 1995 Sep 1;67(17):3069-74
references.pmid: 8779424