Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/50287
標題: Purification of capsid-like particles of infectious bursal disease virus (IBDV) VP2 expressed in E-coli with a metal-ion affinity membrane system
作者: Chen, C.S.
孫幸宜
Suen, S.Y.
Lai, S.Y.
Chang, G.R.L.
Lu, T.C.
Lee, M.S.
Wang, M.Y.
王敏盈
關鍵字: recombinant
protein
protection
vaccines
chromatography
separation
chickens
segment
期刊/報告no:: Journal of Virological Methods, Volume 130, Issue 1-2, Page(s) 51-58.
摘要: Protein VP2, matured from the polyprotein encoded by the genome of infectious bursal disease virus (IBDV) and the primary host-protective immunogen of this virus, together with its two N-terminal truncated mutants were cloned and expressed in Escherichia coli. To obtain pure recombinant proteins for the development of an efficient subunit vaccine against IBDV infection, these three proteins were fused with six additional histidine residues at their C-termini as a His-purification-tag. Following purification employing immobilized metal-ion (Ni2+) affinity chromatography (IMAC), a purification fold of approximately 104 was achieved. Electron microscopic observation also demonstrated that all three E. coli-derived proteins form the morphology of icosahedral particles of approximately 25 nm in diameter. To reduce the cost of resin used for IMAC, self-prepared immobilized metal-ion affinity membranes (IMAM), i.e., commercial, regenerated cellulose membrane modified with iminodiacetic acid and immobilized with nickel ions, was applied to purify particles formed by these three proteins. A 104-fold of purification efficiency was also achieved by this membrane, showing that under the same conditions the recovery and purification efficiency of IMAM are comparable with those of IMAC. The pure VP2-formed particles thus obtained, coupled with their uniform dimensions, not only facilitate a better understanding of the structural biology of these immunogenic particles but also help the development of improved vaccines against this avian virus. (c) 2005 Elsevier B.V. All rights reserved.
URI: http://hdl.handle.net/11455/50287
ISSN: 0166-0934
文章連結: http://dx.doi.org/10.1016/j.jviromet.2005.06.002
Appears in Collections:生物科技學研究所

文件中的檔案:

取得全文請前往華藝線上圖書館



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.