Synthesis and immunochemical properties of the recombinant major surface glycoprotein E2 of the classical swine fever virus
Krivonos, A.V.; Zaberezhny, A.D.; Grebennikova, T.V.; Gibadulin, R.A.; Musienko, M.I.; Tsibezov, V.V.; Bogdanova, V.S.; Kal'nov, S.L.; Aliper, T.I.; Nepoklonov, E.A.
Voprosy Virusologii 45(2): 29-36
2000
ISSN/ISBN: 0507-4088 PMID: 10765548 Document Number: 526131
Recombinant E2 protein from vaccine strain of classical swine fever virus (CSFV) and from SCFV virulent strain Shimen was synthesized in SF-21 and High-Five cell culture with baculovirus as the expressing vector. For secretion, hydrophobic C-terminal transmembrane domain was removed and N-terminal signal polypeptide of 38 amino acids was added. Maximum accumulation of recombinant products in SF-21 cells was observed after 48 h and in medium 96 h after infection with recombinant baculovirus. In High-Five cells and in culture medium the maximum accumulation of E2 was observed after 96 h. The level of E2 expression is 5-10 micrograms/106 cells. The products of expression were purified by affinity chromatography and their specificity confirmed in immunochemical tests with a series of reference monoclonal antibodies. The product can be used for detecting antibodies to SCFV by competitive enzyme immunoassay.