Fast cation-exchange separation of proteins in a plastic microcapillary disc

A novel disposable adsorbent material for fast cation-exchange separation of proteins was developed based on plastic microcapillary films (MCFs). A MCF containing 19 parallel microcapillaries, each with a mean internal diameter of 142 μm, was prepared using a melt extrusion process from an ethylene-...

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Veröffentlicht in:Journal of Chromatography A 2011-03, Vol.1218 (10), p.1409-1415
Hauptverfasser: Darton, N.J., Reis, N.M., Mackley, M.R., Slater, N.K.H.
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Sprache:eng
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Zusammenfassung:A novel disposable adsorbent material for fast cation-exchange separation of proteins was developed based on plastic microcapillary films (MCFs). A MCF containing 19 parallel microcapillaries, each with a mean internal diameter of 142 μm, was prepared using a melt extrusion process from an ethylene-vinyl alcohol copolymer (EVOH). The MCF was surface functionalised to produce a cation-exchange adsorbent (herein referred as MCF-EVOH-SP). The dynamic binding capacity of the new MCF-EVOH-SP material was experimentally determined by frontal analysis using pure protein solutions in a standard liquid chromatography instrument for a range of superficial flow velocities, u LS = 5.5–27.7 cm s −1. The mean dynamic binding capacity for hen-egg lysozyme was found to be approximately 100 μg for a 5 m length film, giving a ligand binding density of 413 ng cm −2. The dynamic binding capacity did not vary significantly over the range of u LS tested. The application of this novel material to subtractive chromatography was demonstrated for anionic BSA and cationic lysozyme at pH 7.2. The chromatographic separation of two cationic proteins, lysozyme and cytochrome-c, was also performed with a view to applying this technology to the analysis or purification of proteins. Future applications might include separation based on anion exchange and other modes of adsorption.
ISSN:0021-9673
1873-3778
DOI:10.1016/j.chroma.2011.01.030