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Title: Fast cation-exchange separation of proteins in a plastic microcapillary disc
Authors: Darton, N.J.
Reis, Nuno M.
Mackley, Malcolm R.
Slater, Nigel K.H.
Issue Date: 2011
Publisher: © Elsevier
Citation: DARTON, N.J. ... et al, 2011. Fast cation-exchange separation of proteins in a plastic microcapillary disc. Journal of Chromatography A, 1218 (10), pp.1409-1415.
Abstract: 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 142μm, 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 =5.5-27.7cms . The mean dynamic binding capacity for hen-egg lysozyme was found to be approximately 100μg for a 5m length film, giving a ligand binding density of 413ngcm . The dynamic binding capacity did not vary significantly over the range of u 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. © 2011 Elsevier B.V.
Description: NOTICE: this is the author’s version of a work that was accepted for publication in Journal of Chromatography A. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published at: http://dx.doi.org/10.1016/j.chroma.2011.01.030
Sponsor: This work was supported by the BBSRC-BRIC scheme and the European Commission (FP7 Marie Curie fellowship scheme).
Version: Accepted for publication
DOI: 10.1016/j.chroma.2011.01.030
URI: https://dspace.lboro.ac.uk/2134/15200
Publisher Link: http://dx.doi.org/10.1016/j.chroma.2011.01.030
ISSN: 0021-9673
Appears in Collections:Published Articles (Chemical Engineering)

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