Characterisation of non-porous magnetic chelator supports and their use to recover polyhistidine-tailed T4 lysozyme from a crude E. coli extract
作者： Susan M O'Brien , Rhona P Sloane , Owen R.T Thomas , Peter Dunnill
DOI: 10.1016/S0168-1656(97)01688-X
关键词: Periplasmic space 、 Sepharose 、 Fusion protein 、 Escherichia coli 、 Selectivity 、 Metal Chelator 、 Chemistry 、 Lysozyme 、 Chelation 、 Chromatography
摘要: The use of high capacity micron-sized non-porous magnetic metal chelator adsorbents for the direct recovery a recombinant metal-binding protein from crude liquors is described. Selectivity and interaction strength particles were assessed using set native proteins with known behaviour towards commercially available immobilised chelate adsorbents. Particles charged Cu2+ highly effective in recovering histidine-tailed T4 lysozyme fusion directly E. coli extracts single step. Levels purity compared favourably those achieved by chromatography pre-clarified on Cu2+-IDA Sepharose. approach offers advantages such as avoidance clarification to prevent fouling columns, steps that become especially significant at large scale. By detailed characterisation chelators practical tailed repeated production periplasmic products realistic prospect.
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