[Synthesis and immunochemical properties of the recombinant major surface glycoprotein E2 of the classical swine fever virus].
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.
Krivonos
A V
AV
Zaberezhny
A D
AD
Grebennikova
T V
TV
Gibadulin
R A
RA
Musienko
M I
MI
Tsibezov
V V
VV
Bogdanova
V S
VS
Kal'nov
S L
SL
Aliper
T I
TI
Nepoklonov
E A
EA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002846:Chromatography, Affinity; D017931:DNA Primers; D007124:Immunoenzyme Techniques; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus