Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/50332
標題: Self-assembly of the infectious bursal disease virus capsid protein, rVP2, expressed in insect cells and purification of immunogenic chimeric rVP2H particles by immobilized metal-ion affinity chromatography
作者: Wang, M.Y.
王敏盈
Kuo, Y.Y.
Lee, M.S.
Doong, S.R.
Ho, J.Y.
Lee, L.H.
關鍵字: infectious bursal disease virus
virus-like particle
baculovirus
expression system
immobilized metal-ion affinity chromatography
genomic segment
protection
chickens
vp2
pathogenicity
antibodies
sequence
antigen
culture
strains
期刊/報告no:: Biotechnology and Bioengineering, Volume 67, Issue 1, Page(s) 104-111.
摘要: A gene encoding a structural protein (VP2) of a local strain (P3009) of infectious bursal disease virus (IBDV) was cloned and expressed using the baculovirus expression system to develop a subunit vaccine against IBDV infection in Taiwan. The expressed rVP2 proteins formed particles of approximately 20-30 nm in diameter. Those particles were partially purified employing sucrose density gradient ultracentrifugation, and the purified particles were recognized by a monoclonal antibody against the VP2 protein of IBDV P3009. To facilitate the purification of the particles, the VP2 protein was engineered to incorporate a metal ion binding site (His)(6) at its C-terminus. The chimeric rVP2H proteins also formed particles, which could be affinity;purified in one step with immobilized metal ions (Ni2+). Particle formation was confirmed by direct observation under the electron microscope. The production level of rVP2H protein was determined to be 20 mg/L in a batch culture of Hi-5 cells by quantifying the concentration of the purified proteins. The chicken protection assay was performed to evaluate the immunogenicity of the rVP2H protein. When susceptible chickens were inoculated with the recombinant rVP2H proteins (40 mu g/bird), virus-neutralizing antibodies were induced, thereby conferring a high level of protection against the challenge of a very virulent strain of IBDV. In conclusion, the most significant finding in this work is that both of the expressed rVP2 and rVP2H proteins can form a particulate structure capable of inducing a strong immunological response in a vaccinated chicken. (C) 2000 John Wiley at Sons, Inc.
URI: http://hdl.handle.net/11455/50332
ISSN: 0006-3592
文章連結: http://dx.doi.org/10.1002/(sici)1097-0290(20000105)67:1<104::aid-bit12>3.0.co
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