Plate height determination for gradient elution chromatography of proteins
Article first published online: 19 FEB 2004
Copyright © 1995 John Wiley & Sons, Inc.
Biotechnology and Bioengineering
Volume 48, Issue 5, pages 444–451, 5 December 1995
How to Cite
Yamamoto, S. (1995), Plate height determination for gradient elution chromatography of proteins. Biotechnol. Bioeng., 48: 444–451. doi: 10.1002/bit.260480506
- Issue published online: 19 FEB 2004
- Article first published online: 19 FEB 2004
- Manuscript Accepted: 31 MAY 1995
- Manuscript Received: 22 NOV 1994
- ion-exchange chromatography;
- protein separation;
- gradient elution;
- preparative separation
A method for determining the plate height HETP from the elution curve obtained by the linear gradient elution (LGE) ion-exchange chromatography (IEC) of proteins is presented. The method was developed on the basis of the numerical solutions of a chromatography model which considers the zone sharpening and the distribution coefficient as a function of the salt concentration. The plate height HETP is determined from the peak width and the salt concentration at which the peak is eluted in LGE. The method was applied to the experimental results with various ion-exchange chromatography media. A calculation example based onthe present method is presented to show how the chromatographic and operating parameters should be tuned to obtain a desired resolution. A simplified calculation procedure for the peak profile is also described. © 1995 John Wiley & Sons, Inc.