Production and characterization of monoclonal antibodies against hog cholera virus (Alfort 187 strain)

Abstract

Summary.A panel of 15 monoclonal antibodies (Mabs) against hog cholera virus (HCV) was produced and each Mab was characterized according to its viral protein specificity, virus-neutralizing activity and immunoreactivity with a large collection of HCV isolates.Hog cholera (HC) is a contagious and economically important disease of pigs and is caused by hog cholera virus (HCV). HCV belongs to the genus pestivirus of the family Flaviviridae [5]. This genus is composed of hog cholera virus, bovine viral diarrhoea virus (BVDV) of cattle and border disease virus (BDV) of sheep. These viruses show strong structural and serological similarities [2,21].Pestiviruses are small enveloped, infectious positive stranded animal RNA viruses with a diameter of about 40 nm, and possess a nucleocapsid assumed to have an icosahedral symmetry [8]. The purification of virions by physical means is extremely difficult [10] due to the problems inherent to pestivirus research, i.e., low titers in cell cultures, membrane association and fragility of viral particles. Despite these purification difficulties, monoclonal antibodies against the HCV were raised in different laboratories. Most of the Mabs produced against the HCV are directed against the major glycoprotein gp 55 [6,18,19], but Mabs were also obtained which recognized the gp44/48 of the Alfort strain and the gp 42 of the "Chinese" vaccine strain [15,20]. Many of the monoclonal antibodies directed against the major glycoprotein were shown to neutralize virus infectivity. Four antigenic domains on the structural gp 55

Characterization of structural and non-structural proteins of hog cholera virus by means of monoclonal antibodies

“…As these monoclonal antibodies did not react in Western blot analyses [4], we could not yet determine against which of these proteins they are directed. By SDS-PAGE electrophoretic analyses in non-denaturing conditions, we observed not only the precipitation of a 65 kDa polypeptide which we assume is the disulfide linked dimer gp 15/16-gp 55, but also a 46 kDa protein formed by the disulfide linked dimer p 27-gp 15/16.…”

“…Caij et al [4] demonstrated that this monoclonal antibody was not HCV restricted, but also recognized BVDV strains. It was previously demonstrated that the non-structural protein p 125 of the cytopathic N A D L strain of BVDV is cleaved to give rise to two proteins, the N-terminal product of 54 kDa and the Cterminal product of 80 kDa [6,19].…”

“…Fifteen monoclonal antibodies (Mabs) were produced by using the ultracentrifuged culture fluid of the HCV strain Alfort 187 as antigen [4].…”