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Separations 2016, 3(4), 31; doi:10.3390/separations3040031

Comparison in Partition Efficiency of Protein Separation between Four Different Tubing Modifications in Spiral High-Speed Countercurrent Chromatography

1
Laboratory of Bioseparation Technology, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
2
Machine Instrumentation Design and Fabrication, National Institutes of Health, Bethesda, MD 20892, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Alain Berthod
Received: 7 July 2016 / Revised: 30 September 2016 / Accepted: 11 October 2016 / Published: 14 October 2016
(This article belongs to the Special Issue Countercurrent Chromatography)
View Full-Text   |   Download PDF [948 KB, uploaded 14 October 2016]   |  

Abstract

High-speed countercurrent chromatography with a spiral tube assembly can retain a satisfactory amount of stationary phase of polymer phase systems used for protein separation. In order to improve the partition efficiency, a simple tool was fabricated to modify the tubing shapes, and the following four different tubing modifications were made: intermittently pressed at 10 mm width, flat, flat-wave, and flat-twist. Partition efficiencies of the separation column made from these modified tubings were examined in protein separation with an aqueous–aqueous polymer phase system at flow rates of 1–2 mL/min under 800 rpm. The results indicated that the column with all modified tubing improved the partition efficiency at a flow rate of 1 mL/min, but at a higher flow rate of 2 mL/min, the columns made of flattened tubing showed lowered partition efficiency, apparently due to the loss of the retained stationary phase. Among all the modified columns, the column with intermittently pressed tubing gave the best peak resolution. It may be concluded that the intermittently pressed and flat-twist improve the partition efficiency in a semi-preparative separation, while other modified tubing of flat and flat-wave configurations may be used for analytical separations with a low flow rate. View Full-Text
Keywords: spiral high-speed countercurrent chromatography; tubing shape modification; protein separation; polymer phase system spiral high-speed countercurrent chromatography; tubing shape modification; protein separation; polymer phase system
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Ito, Y.; Clary, R. Comparison in Partition Efficiency of Protein Separation between Four Different Tubing Modifications in Spiral High-Speed Countercurrent Chromatography. Separations 2016, 3, 31.

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