Premium
Expression, Purification, and Characterization of the Infectious Bursal Disease Virus‐Like Particles Produced by Insect Cells
Author(s) -
Ho JinYi,
Lai SuYuan,
Lee LongHuw,
Wang MinYing
Publication year - 1999
Publication title -
journal of the chinese chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.329
H-Index - 45
eISSN - 2192-6549
pISSN - 0009-4536
DOI - 10.1002/jccs.199900102
Subject(s) - infectious bursal disease , sf9 , capsid , virus , virology , recombinant dna , virus like particle , complementary dna , chemistry , microbiology and biotechnology , monoclonal antibody , antibody , biology , gene , spodoptera , virulence , biochemistry , immunology
The cDNA of the large genomic segment A (ORF A1) of infectious bursal disease virus (IBDV) strain P3009 was synthesized and cloned into the transfer vector pBlueBac4. Following cotransfection and plaque purification, a plaque‐pure recombinant baculovirus, vIBDVJ6, was selected to express IBDV capsid proteins. When Sf9 cells were infected with vIBDVJ6, IBDV precursor protein (polypeptide of VP2‐VP4‐VP3) was expressed and proteolytically processed to give VP2, VP3, and VP4 proteins by a viral protease. These resulting recombinant proteins can self‐assemble to form virus‐like particles (VLPs). The shape and size of the negatively stained purified IBDV‐like particles were demonstrated to be similar to those of the complete P3009 IBDV particles under direct electron microscopic observation. Furthermore, the purified particles were strongly recognized by an anti‐VP2 monoclonal antibody as confirmed by an immuno‐gold labeling experiment. IBDV‐like particles will offer a unique opportunity to create completely non‐infectious viral vaccines for use in the vaccination of chickens. The immunologic response in chickens stimulated by IBDV‐like particles is currently being evaluated.