Premium
Chimeric infectious bursal disease virus‐like particles expressed in insect cells and purified by immobilized metal affinity chromatography
Author(s) -
Hu YuChen,
Bentley William E.,
Edwards Gerard H.,
Vakharia Vikram N.
Publication year - 1999
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19990620)63:6<721::aid-bit10>3.0.co;2-o
Subject(s) - affinity chromatography , antigenicity , infectious bursal disease , virus like particle , virus , recombinant dna , monoclonal antibody , virology , sf9 , capsid , histidine , chemistry , microbiology and biotechnology , fusion protein , spodoptera , antibody , biology , biochemistry , amino acid , enzyme , gene , virulence , immunology
Chimeric virus‐like particles (VLPs) of infectious bursal disease virus (IBDV) were produced by coinfecting Spodoptera frugiperda (S f ‐9) insect cells with two recombinant baculoviruses, vIBD‐7 and vEDLH‐22. vIBD‐7 encodes VP2, VP3, and VP4 of the IBDV structural proteins. vEDLH‐22 encodes VP2 with five histidine residues at the carboxy‐terminus (VP2H). Coinfection produced hybrid VLPs composed of VP2, VP2H, and VP3. The additional histidine residues on VP2H enabled the efficient purification of VLPs based on immobilized metal affinity chromatography (IMAC). These results demonstrated that the VLPs formed are comprised of chimeric subunits with attached affinity ligands, and further, that sufficient His 5 ligand was available for binding to the IMAC metal‐chelating resin. Additionally, these novel particles were fully characterized for antigenicity by a series of monoclonal antibodies, and appeared identical to the two wild‐type IBDV strains contributing subunits to the chimeric VLP. IMAC purification provides a promising low‐cost and simple scheme to purify VLPs as vaccines. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 721–729, 1999.