## Abstract Although it is commonly believed that a column packing used for chromatofocusing must have an “even” buffering capacity in order to produce a linear pH gradient, it is demonstrated here that linear pH gradients suitable for chromatofocusing can be produced on a column packing having a m
Comparison of linear gradient and displacement separations in ion-exchange systems
✍ Scribed by V. Natarajan; S. Ghose; S. M. Cramer
- Publisher
- John Wiley and Sons
- Year
- 2002
- Tongue
- English
- Weight
- 187 KB
- Volume
- 78
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Abstract
The linear gradient mode of chromatography is the most widely employed mode of operation in ion‐exchange chromatographic separations. However, in recent years, the displacement mode has received considerable attention because of its promise of high throughput and high resolution. To enable a comparison of these two modes of chromatography, it is essential to identify the optimum operating conditions for each. We employed an iterative algorithm to carry out the necessary optimization. The Steric Mass Action model of ion‐exchange chromatography is used in concert with the solid‐film linear‐driving force model to describe the chromatographic behavior of the solutes in these systems. The performances of displacement and gradient modes of chromatography are compared for different types of separation problems. It turns out that for “easy” separations, both the modes are equally effective. However, for challenging separations, the displacement mode is superior to the gradient mode. Our results shed significant light on the performance of gradient and displacement modes in protein ion‐exchange systems. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 365–375, 2002.
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