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標題: Purification of VP3 protein of infectious bursal disease virus using nickel ion-immobilized regenerated cellulose-based membranes
作者: Hu, H.L.
Wang, M.Y.
Chung, C.H.
Suen, S.Y.
關鍵字: infectious bursal disease virus;immobilized metal ion affinity;chromatography;immobilized metal ion affinity membrane;VP3 protein;metal affinity membranes;escherichia-coli;antigenic properties;chromatography;oligomerization;adsorption;separation;stabilization;particles;ibdv
Project: Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
期刊/報告no:: Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, Volume 840, Issue 2, Page(s) 76-84.
In this study, hexa-histidine tagged VP3 protein of infectious bursal disease virus (IBDV) was purified using immobilized metal ion affinity technique from the fermentation of Escherichia coli BL21 (DE3) containing a recombinant plasmid with a VP3 gene. The purification efficiencies of VP3 protein (TVP3 and Delta TVP3) using Ni2+-NTA commercial agarose gels and Ni2+-IDA regenerated cellulose-based membranes at 4 degrees C were compared. A good washing condition for removing most impurity proteins was found as 20 mM NaH2PO4, 500 mM NaCl, 40 mM imidazole, pH 7.8, whereas an efficient elution condition was 20 mM NaH2PO4, 500 mM NaCl, 500 or 750 mM[ imidazole, pH 7.8. By applying these conditions to the flow experiments, similar recovery (86-88%) and purity (98-99%) for VP3 were obtained in both gel column (1 ml gel) and membrane cartridge (four membrane disks) under the flow rate of 1.7 ml/min for protein loading and 2.7 ml/min for protein elution. Regarding that the membrane process exhibited some advantages such as shorter residence time and lower cost, a better process efficiency in a large-scale system could be expected for the Ni2+-IDA membranes. (c) 2006 Elsevier B.V. All rights reserved.
ISSN: 1570-0232
DOI: 10.1016/j.jchromb.2006.04.032
Appears in Collections:生物科技學研究所

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