Displacement chromatography of chemotactic peptides |
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Affiliation: | 1. Research Institute, JCR Pharmaceuticals Co., Ltd., Kobe 651-2241, Japan;2. R&D Platform Technology and Science, GlaxoSmithKline, King of Prussia, Pennsylvania 19406;3. R&D Platform Technology and Science, GlaxoSmithKline, Stevenage, UK;1. Protein Research Center, Shahid Beheshti University, G. C., Tehran, Iran;2. Department of Biomedical Engineering, The University of Akron, Akron, OH 44325, USA |
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Abstract: | Displacement chromatography was successfully used to separate a binary peptide mixture, n-formyl-Met–Phe and n-formyl-Met–Trp, on a reversed-phase column. Displacement parameters such as choice of displacer, displacer concentration, mobile phase organic level, and flow-rate were critically examined in the context of maximizing productivity. Since the feed composition was limited by solubility, optimal productivity was sought as a function of feed volume. The impurities contained in the commercial displacers used in this study did not seem to affect the overall separation quality. In most cases the final pattern of contiguous rectangular bands was not attained; nevertheless, separations of high productivity were achieved using benzethonium chloride and tris[2-(2-methoxyethoxy)ethyl]amine as displacers. In some cases further increase in productivity was not possible only because of solubility constraints. Loading of feed at low initial organic modulator level coupled with displacements at higher modulator level was found to give efficient separations. |
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