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標題: Vaccine development through terminal deletions of an infectious bursal disease virus protein 2 precursor variant
作者: Ho, J.Y.
Lee, L.H.
Lin, Y.C.
Tai, Y.J.
Chang, C.K.
Chou, Y.M.
Lai, S.Y.
Wang, M.Y.
關鍵字: Infectious bursal disease virus;Subviral particle;Baculovirus;expression system;Immobilized metal-ion affinity chromatography;Vaccine;ion affinity-chromatography;vp2 subviral particle;capsid protein;insect cells;crystal-structure;chicken il-2;strains;purification;protection;identification
Project: Process Biochemistry
期刊/報告no:: Process Biochemistry, Volume 45, Issue 5, Page(s) 786-793.
VP2 is the primary host-protective immunogen of infectious bursal disease virus (IBDV), the agent that causes the highly contiguous infectious bursal disease (IBD). Previous studies have shown that a C-terminal his-tagged 452 amino acid residue VP2 precursor variant (VP2-452H) can form an immunogenic subviral particle (SVP). A set of his-tagged N- and C-terminal VP2-452 deleting mutants (designated as N5-452H, N10-452H, N20-452H, N40-452H, VP2-441H, VP2-437H, VP2-411H and VP2-399H) was expressed in insect cells to discover the role of both N- and C-termini on the assembly of SVP and to develop an efficient SVP-based vaccine. Among these mutants, the expression level of N5-452H was the highest. Results of ultracentrifugation and electron microscopy also indicated that mutants of N-terminal deletion N10-452H, N20-452H and N40-452H or C-terminal deletion VP2-411H and VP2-399H lost the capability to self-assemble SVP. The other mutants, N5-452H, VP2-441H and VP2-437H, formed SVP. Additionally, SVP formed by N5-452H could not only be single-step purified by immobilized metalion affinity chromatography (IMAC), but it could also induce a high titer of neutralizing activity to protect chicks from the infection of IBDV at a low dosage (0.2 mu g), suggesting that SVP formed by N5-452H can be an alternative vaccine candidate for the prevention of IBD. (C) 2010 Elsevier Ltd. All rights reserved.
ISSN: 1359-5113
DOI: 10.1016/j.procbio.2010.01.021
Appears in Collections:生物科技學研究所

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