期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1272, 期 -, 页码 145-149出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2012.11.036
关键词
Megaporous cryogels; Immobilized metal-ion affinity; Chromatography; Hydrodynamics; Dynamic binding capacity
资金
- Higher Education Commission of Pakistan
- European Commission [PGSYN/ETB-2008-44]
Megaporous cryogels with metal-ion affinity functionality, which possess enhanced protein-binding ability, were synthesized and their properties were investigated. These highly porous materials (pore sizes up to 100 mu m) allowed the direct capture of a recombinant His(6)-tagged protein from a partially clarified extract. The total ligand density of the material was found to be 770 mu mol/g. Application of a partially clarified cell extract in order to recover a His(6)-tagged protein (NAD(P)H-dependent 2-cyclohexen-1-one-reductase) yielded 12 mg of highly purified recombinant product per gram of adsorbent. Increased dynamic binding capacities were observed upon larger degrees of grafting, although some reduction in the quality of the system hydrodynamics was also observed. Nevertheless, these immobilized metal-ion affinity cryogels show potential for a convenient single-step purification of recombinant proteins from raw cell extracts without the need for laborious pre-chromatographic sample clean-up procedures. (C) 2012 Elsevier B.V. All rights reserved.
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