Application of a beta-cyclodextrin sulfate-immobilized precolumn to selective on-line enrichment and separation of heparin-binding proteins by column-switching high-performance liquid chromatography

Journal of Chromatography. a
K IshimuraH Nakamura

Abstract

A column-switching high-performance liquid chromatography (HPLC) system which consisted of a beta-cyclodextrin (beta-CD) sulfate-immobilized hydrophilic vinyl-polymer gel precolumn and a reversed-phase analytical column was developed for the selective on-line enrichment and separation of heparin-binding proteins. Of 15 proteins investigated, 10 proteins having heparin-binding activity were retained on the beta-CD sulfate precolumn almost quantitatively, in contrast 5 proteins having no heparin-binding activity were not retained. Calibration graphs for basic fibroblast growth factor constructed at various sample volumes were nearly identical, indicating that the protein could be enriched by this system. The system was successfully used for the selective separation of lysozyme in egg white. The beta-CD sulfate-immobilized precolumn showed no loss of analytical performance over 2 years during which about 400 samples were analysed.

Citations

Jun 6, 2000·Advanced Drug Delivery Reviews·T Irie, K Uekama
Aug 19, 2007·Journal of Virology·Marguerite PriorHilary E M McMahon
Feb 21, 2013·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Ji Sheng Yang, Li Yang
Jul 13, 2000·Journal of Chromatography. a·L Bovanová, E Brandsteterová
Feb 19, 2002·Chemical Reviews·Kaneto UekamaTetsumi Irie

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